Ahmad Et Al., (2020) Molecular Mwchanisms of Adipogenesis

REVIEW
published: 08 May 2020

doi: 10.3389/fmolb.2020.00076
Molecular Mechanisms of
Adipogenesis: The Anti-adipogenic
Role of AMP-Activated Protein
Kinase
Bilal Ahmad 1 , Christopher J. Serpell 2 , Isabel Lim Fong 3 and Eng Hwa Wong 4*
1
School of Biosciences, Faculty of Health and Medical Sciences, Taylor’s University, Subang Jaya, Malaysia, 2 School
of Physical Sciences, University of Kent, Canterbury, United Kingdom, 3 Department of Paraclinical Sciences, Faculty
of Medicine and Health Sciences, Universiti Malaysia Sarawak, Kota Samarahan, Malaysia, 4 School of Medicine, Faculty
of Health and Medical Sciences, Taylor’s University, Subang Jaya, Malaysia
Obesity is now a widespread disorder, and its prevalence has become a critical
concern worldwide, due to its association with common co-morbidities like cancer,
cardiovascular diseases and diabetes. Adipose tissue is an endocrine organ and
therefore plays a critical role in the survival of an individual, but its dysfunction or excess
is directly linked to obesity. The journey from multipotent mesenchymal stem cells to
the formation of mature adipocytes is a well-orchestrated program which requires the
Edited by: expression of several genes, their transcriptional factors, and signaling intermediates
Eleonora Napoli,
University of California, Davis,
from numerous pathways. Understanding all the intricacies of adipogenesis is vital if we
United States are to counter the current epidemic of obesity because the limited understanding of
Reviewed by: these intricacies is the main barrier to the development of potent therapeutic strategies
José María Moreno-Navarrete, against obesity. In particular, AMP-Activated Protein Kinase (AMPK) plays a crucial role
CIBER Fisiopatología Obesidad y
Nutrición (CIBEROBN), Spain in regulating adipogenesis – it is arguably the central cellular energy regulation protein
Rubén Cereijo, of the body. Since AMPK promotes the development of brown adipose tissue over that
University of Barcelona, Spain
of white adipose tissue, special attention has been given to its role in adipose tissue
*Correspondence:
Eng Hwa Wong
development in recent years. In this review, we describe the molecular mechanisms
EngHwa.Wong@taylors.edu.my involved in adipogenesis, the role of signaling pathways and the substantial role of
activated AMPK in the inhibition of adiposity, concluding with observations which will
Specialty section:
This article was submitted to
support the development of novel chemotherapies against obesity epidemics.
Cellular Biochemistry,
Keywords: obesity, adipogenesis, WAT, BAT, AMPK, beige/brite adipocytes
a section of the journal
Frontiers in Molecular Biosciences
Received: 10 February 2020 INTRODUCTION
Accepted: 03 April 2020
Published: 08 May 2020
Obesity is an increasingly prevalent disorder around the globe promoted by genetic, nutritional,
Citation: and environmental factors. Energy imbalance – excessive consumption of calories compared to
Ahmad B, Serpell CJ, Fong IL and utilization is the key driving force of obesity. Obesity is a multifactorial chronic disease, linked to
Wong EH (2020) Molecular
other disorders including cancer, insulin resistance, cardiovascular diseases and type-2 diabetes
Mechanisms of Adipogenesis:
The Anti-adipogenic Role
(Cohen et al., 2014; Tung et al., 2017; Ghaben and Scherer, 2019). Obesity/overweight is now
of AMP-Activated Protein Kinase. the fifth leading cause of death worldwide (Chandrasekaran et al., 2012). According to World
Front. Mol. Biosci. 7:76. Health Organization (WHO), in 2016, more than 1.9 billion adults were overweight, of whom 650
doi: 10.3389/fmolb.2020.00076 million were suffereing from obesity, with approximately 2.8 million adults dying each year due
Frontiers in Molecular Biosciences | www.frontiersin.org 1 May 2020 | Volume 7 | Article 76

Ahmad et al. Molecular Opportunities in Adipogenesis Intervention
to the condition (World Health Organization, 2020). There are ADIPOSE TISSUE AND ADIPOGENESIS
many factors which contribute to obesity such as sedentary
lifestyle, high calorific intake, depression, and various social and Adipose tissue is one of the most complex organs in the
monetary issues, but they have a single common result: the human body, containing pre-adipocytes, adipocytes, immune
accumulation of fats in mature adipocytes of white adipose tissue cells, fibroblasts, pericytes, vascular smooth muscle cells, and
(WAT): obesity is characterized by increase in the mass of adipose vascular endothelial cells (Bijland et al., 2013; Coelho et al., 2013;
tissue (Ghaben and Scherer, 2019). The epidemic of obesity has Schling and Löffler, 2018). At the whole body level, adipose tissue
therefore focused researchers’ attention on understanding the is divided into visceral adipose tissue (VAT) and subcutaneous
development of adipose tissue and fat cells, regulated by a multi- adipose tissue (SCAT) (Bijland et al., 2013). The tissue secretes
step process known as adipogenesis (Lefterova and Lazar, 2009). various adipokines and has regulatory roles in the endocrine,
Improved and holistic knowledge of the processes governing immune and metabolic systems (Bijland et al., 2013; Booth
adipogenesis is required if we are to counter the burgeoning et al., 2015). WAT is the hub for the synthesis and storage of
epidemic (Rosen and MacDougald, 2006). triglycerides. Maintenance of systematic energy balance through
Adipose tissue plays an important proper role in the body. storage and release of free fatty acids and via secretion of
Excess energy is stored in the form of fat, in mature adipocytes adipokines is the main function of WAT (Raajendiran et al.,
within WAT (of which these cells make up the majority) (Niemalä 2019). Whether located viscerally or subcutaneously, adipose
et al., 2008), and during energy scarcity, these fats are used by tissue has a crucial role in the survival of an individual because
other organs of the body to meet the energy demand (Moseti it is the basic source of fatty acids for the production of heat
et al., 2016). Obesity is diagnosed either on the amount of WAT and energy. White adipocytes or white fat cells are lipid-laden
or the number of mature white adipocytes in WAT (Park, 2014), cells within WAT that acquire the ability to accumulate lipids
rather than simply by body weight. The expansion of adipose after differentiation – the process in which the cells from a
depots can be driven either by the increase in the number common ancestor are derived mitotically and become different
(hyperplasia) or size (hypertrophy) of adipocytes (Ghaben and from one another in morphology and function. Adipocytes are
Scherer, 2019). Both hyperplasia and hypertrophy are responsible derived from multipotent mesenchymal stem cells (MSCs), which
for the dysfunctionality of adipose tissue (Unamuno et al., are first transformed into pre-adipocytes before undergoing
2018). Changes in the physiological functions of dysfunctional secondary differentiation to become mature adipocytes. The
adipose tissue (such as abnormal secretion of adipokines, insulin differentiation of adipocytes is determined by the expression of
resistance, and chronic inflammation) are directly linked to genes and the function of proteins which dictate the phenotype
obesity and its related co-morbidities (Van Kruijsdijk et al., of adipocytes (Ali et al., 2013). Hyperplasia and hypertrophy
2009). For example, inflammation in adipose tissue due to of WAT through adipogenesis (e.g., due to excessive energy
abnormal secretion of proinflammatory adipokines such as intake accompanied by low energy expenditure) leads to obesity
monocyte chemoattractant protein-1 (MCP-1), tumor necrosis (Tang and Lane, 2012; Ali et al., 2013; Haider and Larose,
factor-alpha (TNFα), Interleukin 6 (IL 6) etc. is a key factor in 2019). The cellular process of adipogenesis involves three well-
the development of type 2 diabetes (T2D), insulin resistance, defined stages: (i) commitment of MSCs to the adipocyte lineage;
cardiovascular diseases and cancer (Burhans et al., 2011; Park (ii) mitotic clonal expansion - involving replication of DNA
et al., 2014). So, therefore it is essential to understand the and duplication of cells; (iii) terminal differentiation, involving
molecular mechanisms of adipocyte differentiation, physiology, expression of genes and transcriptional factors such as CCAAT/
and morphology of adipocytes in order to contribute to the enhancer-binding proteins (C/EBPs) family and peroxisome
overall understanding of adiposity and develop new ways to proliferator-activated receptor–γ (PPARγ) – and a dramatic
combat obesity and its associated complications. A concise increase in lipogenesis and induction of lipogenic genes such
knowledge of the role of genes, proteins (transcriptional as acetyl CoA carboxylase (ACC), Fatty acid synthase (FAS)
factors and hormones), and signaling intermediates regulating and adipocyte fatty acid binding protein (aP2) (Lefterova and
adipogenesis is also of vital importance and all of these Lazar, 2009; Khalilpourfarshbafi et al., 2018; Lazar et al., 2018).
factors are promising targets for the discovery of novel anti- The differentiation of pre-adipocytes into mature adipocytes is
obesogenic drugs. In particular, the role of energy sensor also influenced by various other factors including growth factors
protein of the body, adenosine monophosphate-activated protein such as insulin-like growth factor 1 (IGF-1) (Lefterova and
kinase (AMPK, a negative regulator of white adipogenesis) in Lazar, 2009), and insulin itself. IGF-1 is critical for the survival,
adipose tissue development is of central importance as we will proliferation and differentiation of pre-adipocytes (Garten et al.,
argue in this review. 2012), and insulin is one of the potent adipogenic hormones,
This review aims to provide a comprehensive knowledge inducing the transcription of various positive regulators of
of the key molecular factors (proteins and various signaling adipogenesis (Klemm et al., 2001). During adipogenesis, the cells
pathways) involved in adipocyte differentiation, and the anti- lose their fibroblastic shape and become spherical, which is an
adipogenic role of AMPK and its mechanism of activation in indication of profound changes taking place in the extracellular
adipose tissue. But before heading toward those details, we must matrix (ECM), and cytoskeletal components of the cells (Niemalä
survey the origin, physiology and types of adipose tissues, and the et al., 2008), including decreased expression of actin (Lazar et al.,
biomolecules responsible for the morphological changes during 2018). Alteration in the organization of actin may influence the
adipocyte differentiation. cytoskeletal tension, something which has been shown to regulate

adipogenesis in vitro (Schiller et al., 2013). Various components giving what is known as beige or ‘brite’ (brown in white)
of the ECM, negatively or positively regulate the differentiation adipocytes. Zfp516 is a novel transcriptional activator of UCP1
of pre-adipocytes (Sarantopoulos et al., 2018). For instance, and can be induced by hormonal stimulation, exposure to cold,
proteolytic degradation of the ECM around pre-adipocytes and innervation (Dempersmier et al., 2015). It directly binds
by a cascade of plasminogen is essential for changes in the to the proximal region of UCP1 promoter and interacts with
expression of adipogenic genes and deposition of fats (Selvarajan transcriptional co-regulator PR-domain containing 16 (PRDM
et al., 2001; Ali et al., 2013). Selvarajan et al. (2001) reported 16) to activate UCP1 promoter (Dempersmier et al., 2015). In
that events and changes (molecular and morphological) which addition to Zfp516, various other transcriptional regulators have
were associated with these changes in ECM might modulate also been implicated in the activation of brown/beige adipocytes
adipogenesis directly because they alter the expression of positive specific genes (Shapira and Seale, 2019). These include interferon
transcriptional regulators of adipogenesis such as PPARγ and regulatory factor 4 (IRF4), Krüppel-like factor 11 (KLF11), TATA-
C/EBPα. The expression of another protein, preadipocyte factor- binding protein associated factor 7L (TAF7L), zinc finger and
1 (PREF-1), which is considered to be responsible for maintaining BTB domain-containing protein 16 (ZBTB16), placenta-specific
the phenotype of pre-adipocytes, decreases dramatically upon gene 8 protein (PLAC8), early B cell factor 2 (EBF2), forkhead
induction of adipocytes differentiation (Lazar et al., 2018). box C2 (FoxC2) and ewing sarcoma break point region 1
Each year, approximately 10% of adipocytes turn over in (EWSR1) (Seale et al., 2009; Seale, 2015; Inagaki et al., 2016;
human adipose tissue (Lowe et al., 2011). This long duration Mueller, 2016; Shapira and Seale, 2019). Beige adipocytes are
means that the proper functioning of these newly formed inducible and possess characteristics of both WAT and BAT
adipocytes must be ensured to prevent dysfunction and metabolic (Herz and Kiefer, 2019). Under basal conditions, beige adipocytes
diseases (Lowe et al., 2011). Promotion of normal function of in WAT show phenotypes similar to white adipocytes: they
adipocytes, or replacement of poorly functioning adipocytes may lack expression of UCP1 and contain one large lipid droplet
prove beneficial in overcoming the problem of obesity and its (Petrovic et al., 2010). However, when exposed to cold (Barbatelli
associated disorders. et al., 2010) and β3-adrenergic activators (Himms-Hagen et al.,
2000), these beige cells acquire characteristics similar to brown
adipocytes including expression of UCP1 and presence of small
BIOLOGY OF WHITE, BROWN AND multilocular lipid droplets. Recruitment of beige adipocytes
BEIGE ADIPOSE TISSUES within WAT leads to the acquisition of thermogenic capacity
in WAT, just like that of BAT (Chayama et al., 2019). Beige
There are two main types of adipose tissues in mammals; adipocytes can also be differentiated de novo from the dedicated
white and brown adipose tissues (WAT and BAT), characterized white precursor cells, whenever stimuli such as β3-adrenergic
by different morphologies, anatomical locations, biochemical activators or exposure to cold are met (Figure 1) (Harms and
features, functions and gene expression patterns. Both are Seale, 2013; Rosen and Spiegelman, 2014; Merlin et al., 2016).
involved in the homeostasis of energy (Park, 2014). The main However, they are converted back to white adipocytes when
constituent of adipose tissue is WAT, which is used as an heat generation is no longer a priority (Rosen and Spiegelman,
energy substrate when needed. WAT adipocytes have a greater 2014), illustrating that these cells exhibit extraordinary plasticity
average diameter (20–150 µm) than those of BAT (10–25 µm) in response to changes in the physiological conditions. The
(Stock and Cinti, 2003). White adipocytes contain a single thermogenic activity of beige cells has been reported to act
lipid droplet of triglycerides (formed from esterification of fatty against obesity and increase energy expenditure (Crane et al.,
acids and glycerol-3-phosphate). WAT represents more than 2015). The prevalence of beige cells is in inverse proportion to
95% of adipose mass while BAT represents 1–2% of the fat obesity, body mass index, and plasma glucose level (Cypess et al.,
(Kahn et al., 2019). Brown adipocytes contain high numbers 2009), evidencing the importance of their role in the regulation
of multilocular lipid droplets as well as many mitochondria of body’s metabolism. Some reports had stated that BAT was
(Park, 2014). BAT is known to be protective against hypothermia present only in newborns and small mammals, but recent studies
due to its capacity to break down lipids to generate heat have revealed conclusively the presence and functional relevance
(thermogenesis). WAT stores triglycerides while BAT disperses of BAT (Saely et al., 2011) and beige adipose tissue in adults
energy in thermogenesis - thus there is a complementary (Shinoda et al., 2015).
functional relationship between the two forms (Coelho et al., Although both brown and white adipocytes originate from
2013; Mukherjee et al., 2015). Mitochondria within BAT host key MSCs, it is believed that the immediate precursor cells giving
thermogenic protein uncoupling protein 1 (UCP1), which is a rise to brown and white adipocytes are different (Figure 1):
crucial player for thermogenesis (Tam et al., 2012; Shan et al., MSCs are pledged either to adipogenic (Myf-5 negative) cells
2016). UCP1 is expressed in the inner membrane of mitochondria giving rise to white adipocytes, or myogenic (Myf-5 positive)
and is responsible for the generation of heat via respiratory cells that become brown adipocytes (Timmons et al., 2007;
uncoupling reactions. It converts chemical energy into heat Park, 2014). The Myf-negative precursor cells give rise to white
via proton leak across the inner membrane of mitochondria pre-adipocytes through the expression of bone morphogenetic
(Park, 2014). The expression of UCP1 in WAT has also protein-2 and 4 (BMP 2 and 4) and beige pre-adipocytes
been reported previously: over-expression of the transcriptional upon exposure to cold or β3-adrenergic activators. BMP 2
activator (Zfp516) of UCP1 resulted in the ‘browning’ of WAT, is known to promote osteogenesis in human bone-marrow

FIGURE 1 | Differentiation of MSCs into white, beige/brite and brown adipocytes. Myf-5, Myogenic Factor-5 protein; BMP 7, Bone morphogenetic protein 7; BMP 2,
Bone morphogenetic protein 2; BMP 4, Bone morphogenetic protein 4; PRDM 16, PR-domain containing 16; PGC-1α, peroxisome proliferator-activated receptor
gamma coactivator 1-alpha; PPARγ, peroxisome proliferator-activated receptor gamma; PTEN, phosphatase and tensin homologue; C/EBPα,β,δ,
CCCAAT/Enhancer Binding Protein α, β, δ.
cells and white adipogenesis in mouse-derived 3T3-L1 and in the differentiation of both WAT and BAT but with binding
C3H10T1/2 cells by inducing the expression of PPARγ (Margoni sites specific to either WAT or BAT. For example, early beta-
et al., 2012). BMP 4 promotes the commitment of MSCs to cell factor-2 and PRDM 16 recruits PPARγ to BAT selective
the adipogenic lineage and is reported to induce adipogenesis genes, while TLE3 recruits PPARγ to activate specifically white
in a dose-dependent manner in mouse embryonic stem cells adipogenesis (Giralt and Villarroya, 2013).
(Taha et al., 2006). The Myf-positive precursor cells give rise The recent rediscovery of effective BAT in adult humans
to brown pre-adipocytes through bone morphogenetic protein- has invigorated interest in it as a viable and novel target
7 (BMP 7) expression and PRDM16 (Kajimura et al., 2009; for anti-obesogenic drugs (Park, 2014). BAT transplantation
Park, 2014). BMP 7 activates brown adipogenesis only through studies have revealed that besides thermogenic activities,
the p38MAPK pathway by inducing the expression of brown BAT also acts as an endocrine organ and secretes various
adipogenesis-specific transcriptional factors such as UCP1, brown adipokines known as batokines to orchestrate adaptive
Peroxisome proliferator-activated receptor gamma coactivator thermogenesis and, in turn, improving metabolic health (White
1-alpha, beta (PGC-1α,β), C/EBPs and PPARγ (Tseng et al., et al., 2019). These batokines exert endocrine, autocrine and
2008). PRDM16 is expressed both before and after adipocyte paracrine actions and target distant organs to exert their
differentiation (Morganstein et al., 2010). Both PRDM16 and effects (Villarroya et al., 2019). Batokines include fibroblast
C/EBPβ together act as a switch in determining the fate of growth factor-21 (FGF21), C-X-C motif chemokine ligand-14
BAT away from myogenic lineage by inducing the expression (CXCL14), bone morphogenetic protein-8b (BMP8b), growth-
of PGC-1α and PPARγ (Park, 2014). It has also been reported and-differentiation factor-15 (GDF15), neuregulin-4 (NRG4),
that myf expressing cells (myf-5 positive precursor cells) can S100 calcium-binding protein b (S100b) and various others
be differentiated into white adipocytes (Harms and Seale, 2013; (Villarroya et al., 2019). These secreted batokines perform various
Rosen and Spiegelman, 2014). Sánchez-Gurmaches et al. (2012) functions and contribute to the regulation of immune activities,
evidenced a subset of white adipocytes which were derieved thermogenic activities, cardioprotective effects, vascularization,
from myf-5 positive progenitor cells. Loss of phosphatase and substrate utilization etc. (Lee et al., 2019; Villarroya et al.,
tensin homologue (PTEN) in myf-5 positive precursors resulted 2019). The activity of BAT in human is in inverse relation
in a subset of white adipocytes (Sánchez-Gurmaches et al., to the onset of obesity, type II diabetes and age (Lee et al.,
2012). However, most of the evidence supports that BAT and 2013). Upregulating the proteins and transcriptional factors
WAT originates from different developmental paths (Rosen specifically expressed in brown or beige adipocytes is a highly
and Spiegelman, 2014). Recently, efforts have been made to promising approach in the elimination of obesity. Activation
identify the transcriptional mechanisms specific to WAT and of the thermogenic system in humans, either in WAT or BAT,
BAT-related gene regulatory networks. It has been observed that should correlate well with an increase of energy expenditure.
most of the adipogenic factors, for example, PPARγ functions Thus, developing browning-inducing strategies in WAT or

activation of BAT might contribute to a crucial strategy for KLF7 significantly decreases C/EBPα, PPARγ, adipsin and aP2
treating obesity. expression (Kawamura et al., 2006). KLF16 overexpression
inhibits the differentiation of 3T3-L1 cells and brown pre-
adipocytes through downregulation of PPARγ expression, while
TRANSCRIPTIONAL REGULATION OF knockdown of KLF16 promotes differentiation of both brown,
ADIPOGENESIS and white adipocytes and increases expression of PPARγ (Jang
et al., 2016). Similarly, GATA binding proteins 2 and 3 (GATA2
Adipogenesis is controlled by a large number of transcriptional and 3) also decrease the rate of adipogenesis by downregulating
factors, including C/EBP family members and PPARγ (Ali et al., PPARγ expression (Tong et al., 2005; Khalilpourfarshbafi et al.,
2013). Expression of C/EBPβ and C/EBPδ occurs at early stages of 2018). GATA2 and 3 are expressed predominantly in the pre-
adipocyte differentiation and together they induce the expression adipocytes of WAT, decreasing the expression of PPARγ2 –
of C/EBPα and PPARγ which are the central positive modulators and hence adipogenesis - through direct suppression of PPARγ2
of adipogenesis (Rosen et al., 2009; Khalilpourfarshbafi et al., promoter, and formation of inhibitory complexes with C/EBP
2018). C/EBPβ is considered the most important, being induced family members. GATA2 and 3 are also known to have a
rapidly after the induction of adipogenic stimuli (Guo et al., role as molecular gatekeepers during the differentiation of
2015). Knockdown of C/EBPβ is reported to block adipogenesis adipocytes and may be novel targets for preventative anti-
in 3T3-L1 adipocytes (Zhang et al., 2011; Guo et al., 2012, 2015). obesogenic therapies (Feng et al., 2016). Another protein, PREF1,
PPARγ is the master regulator involved in the differentiation is expressed abundantly in pre-adipocytes, but its expression
of adipocytes and metabolism (Lefterova et al., 2014). PPARγ decreases significantly upon the development of adipocytes.
and C/EBPα exert positive feedback on each other (Figure 2), Ectopic expression of PREF1 inhibits 3T3-L1 differentiation
co-operating to orchestrate the complete adipogenic program and reduces the expression of C/EBPα and PPARγ (Rosen and
(Khalilpourfarshbafi et al., 2018). Several studies (Barak et al., MacDougald, 2006; Sarjeant and Stephens, 2012). Mice deficient
1999; Rosen et al., 1999) have indicated that PPARγ is the in PREF1 showed retarded growth and enhanced adiposity
key regulator involved in the development and differentiation (Sarjeant and Stephens, 2012). Other transcriptional factors
of adipocytes, and therefore known to be obligated for the such as cyclic AMP response binding element (CREB) and
differentiation of adipocytes; cells deficient in PPARγ cannot sterol regulatory binding protein-1 (SREBP1), (which expedites
differentiate into mature adipocytes even if other powerful pro- metabolism of fatty acids by inducing expression of PPARγ)
adipogenic factors are ectopically expressed (Rosen et al., 2009). are positive regulators of adipogenesis and needed in the
Previous in vitro studies have shown that most of the activators differentiation of white pre-adipocytes into mature adipocytes
and repressors of adipogenesis alter the activity and expression (Khalilpourfarshbafi et al., 2018). In white pre-adipocytes CREB
of PPARγ (Sarjeant and Stephens, 2012). Transcriptional factors is required for the induction of differentiation of adipocytes and
such as C/EBPβ, C/EBPδ, Kruppel-like factor 5 (KLF5) and absence of CREB inhibits pre-adipocytes differentiation (Reusch
early β-cell factor 1 (EBF1) are known to directly induce the et al., 2000). CREB is required to induce the expression of
expression of PPARγ mRNA in adipogenesis (Rosen et al., C/EBPβ during the early stages of adipocyte differentiation. It
2009). Early β-cell factor 1 and 2 (EBF1 and EBF2) are induced binds to the dual cis regulatory elements (TGA1 and TGA2)
during the differentiation of the 3T3-L1 white pre-adipocyte within the proximal promoter region of C/EBPβ gene and
cell line, but their pattern of expression is different from each activates its transcription. Expression of a dominant-negative
other (Jimenez et al., 2007). EBF1 binds to the promoter of CREB in mouse embryonic fibroblasts (MEFs) has been observed
C/EBPα, directly activating C/EBPα and PPARγ (Jimenez et al., to block adipogenesis and expression of C/EBPβ (Zhang et al.,
2007). EBF2 has been reported to regulate brown adipocyte 2004). SREBP1 is also involved in adipocytes differentiation,
genes expression (Ucp1 and Prdm16) and is expressed at higher may induce the expression of PPARγ and metabolism of fatty
levels in BAT as compared to WAT (Rajakumari et al., 2013). acids (Khalilpourfarshbafi et al., 2018). SREBP-1c is the highly
Reduction of EBF1 and 2 blocks the differentiation of 3T3-L1 cells expressed form of SREB1 in adipocytes (Sewter et al., 2002;
(Jimenez et al., 2007). Payne et al., 2010) and involved in the regulation of genes
Likewise, other transcriptional factors also contribute to the responsible for the synthesis of fatty acids such as FAS (Rosen
regulation of adipogenesis. Kruppel-like factors (KLFs) may et al., 2009). In addition, retinoblastoma protein (pRb) is also
be either activators or suppressors of adipogenesis. KLF4, known to positively regulate white adipogenesis and is the
KLF5, KLF6, KLF9, KLF13 and KLF15 are known to enhance founding member of the pocket proteins family (Hansen et al.,
adipogenesis while KLF2, KLF3, KLF7 and KLF16 inhibit 2004). It is considered as an essential player in the differentiation
adipogenesis (Sue et al., 2008; Jiang et al., 2015; Jang et al., 2016; of white adipocytes in mice (Moreno-Navarrete et al., 2013).
Pollak et al., 2018). Adenovirus-mediated ectopic expression of Activation of pRb positively regulates terminal differentiation
KLF2 has been reported to inhibit the expression of C/EBPα, of white adipocytes and inhibits brown adipogenesis (Chen
PPARγ and sterol regulatory binding protein-1c (SREBP-1c) but et al., 1996; Moreno-Navarrete et al., 2013; Petrov et al., 2015).
did not have any effect on C/EBPδ and C/EBPβ expressions The adipogenic effects of pRB are due to its regulatory effects
(Banerjee et al., 2003). KLF3 inhibits 3T3-L1 pre-adipocyte on memebers of C/EBPs family, especially C/EBPβ. It binds
differentiation by repressing the C/EBPα promoter (Sue et al., and augments the activity of C/EBPβ and hence positively
2008), as does KLF7 (Cho et al., 2007); overexpression of regulates white adipogenesis. pRb-deficient fibroblasts are unable

FIGURE 2 | Transcriptional regulation of adipogenesis. Arrows represent activation and bars represent inhibition. MSCs, mesenchymal stem cells; DNA,
deoxyribonucleic acids; C/EBPs, CCAAT/enhancer binding proteins. C/EBPβ,δ; CCCAAT/Enhancer Binding Protein β, δ, KLF 4,5,6,9,13,15, Kruppel-like factor
4,5,6,9,13,15; SREBP-1, Sterol regulatory binding protein-1; KLF 2,3,7,16, Kruppel-like factor 2,3,7,16; GATA 2,3, GATA binding protein 2,3; EBF-1, 2, Early β-cell
factor 1,2; CREB, cyclic AMP response binding element; PREF-1, preadipocyte factor-1; pRb, retinoblastoma protein; FAS, fatty acid synthase; ACC, acetyl CoA
carboxylase; FABP-4, fatty acid binding protein-4.
to undergo adipose conversion (Hallenborg et al., 2009). pRb also antagonistically with overexpression of one factor repressing the
regulates fate choice and lineage commitment (Calo et al., 2010). other (Zhang et al., 2006; James, 2013).
Lack of pRb switches the cell fate from white to brown adipocyte,
increases energy expenditure and acts as molecular switch in the
determination of white versus brown adipogenesis (Hansen et al., Transforming Growth Factors-β Pathway
2004; Dali-Youcef et al., 2007). Inactivation of pRb in mouse and Bone Morphogenetic Proteins
embryonic fibroblasts, white pre-adipocytes and mouse stem cells The transforming growth factors-β (TGF-β) pathway consists
resulted in increase brown adipogenesis and increased expression of more than 33 members. These include TGF-β1, 2, and 3,
of UCP1 (Hansen et al., 2004) which demonstrates that activation bone morphogenetic proteins (BMPs), activins, nodal-related
of pRb is positively associated with white adipogenesis and its proteins and growth differentiation factors (GDFs) (Lee, 2018).
ablation promotes brown/beige adipogenesis. Members of the TGF-β super family control diverse process
As, both PPARγ and C/EBP family members are the central such as cell differentiation, growth and cell fate specification
modulators of adipogenesis and are widely studied targets in (Budi et al., 2017; Lee, 2018) in various cell types including
in vitro and in vivo studies of anti-obesogenic medicine, so, adipocytes (Margoni et al., 2012). The TGF-β pathway stimulates
insights into various signaling pathways, energy sensing proteins proliferation of pre-adipocytes but inhibits differentiation of pre-
(e.g., AMPK), genes and their transcriptional factors which have adipocytes into mature adipocytes (Zamani and Brown, 2010;
direct interactions with PPARγ and C/EBP family members are Lee, 2018). Among all the TGF-β superfamily members, TGF-β1
required to tackle abnormal adipose tissue development and has the greatest role in adipogenesis, inhibiting 3T3-L1 pre-
obesity pandemic. adipocyte differentiation (Margoni et al., 2012) by interacting and
repressing PPARγ, C/EBPα and C/EBPβ (Moseti et al., 2016). The
signal transduction in TGF-β pathway begins when the TGF- β
ROLE OF SIGNALING PATHWAYS IN ligands bind to type 1 and 2 receptors (TGF-β-R1 and TGF-β-R2)
ADIPOGENESIS present on the cell surface. These receptors are serine/threonine
kinases and convey the signals through downstream processes
MSCs are committed to either osteogenic, myogenic or (Margoni et al., 2012). TGF-β ligands bind to the TGF-β-R2
adipogenic lineages. This involves discrete signaling pathways receptor and recruit (phosphorylate) the TGF-β-R1 receptor. The
including those of Bone morphogenetic protein (BMP), Wnt phosphorylated TGF-β-R1 receptor then targets and potentiates
(canonical and non-canonical), and Hedgehog. These pathways the downstream specific receptor-regulated SMAD proteins
exert very strong influences on the central regulators of referred as R-SMADs. In TGF-β branch of the pathway two
both osteogenesis (Runx2) and adipogenesis (PPARγ), working R-SMADs (SMAD2/3) take part in the process. These R-SMADs

upon phosphorylation associate with common-SMAD (co- in the differentiation of adipocytes depending on the stage
SMAD/SMAD4) and form a heterocomplex. This complex then of cells, BMP type and dosage (Tseng and He, 2007). BMP-
translocates to the nucleus and activates the target genes (Lee, 2 and 4 have been shown to commit pluripotent stem cells
2018) (Figure 3). toward the adipogenic lineage (Tang and Lane, 2012). BMP-
Bone morphogenetic proteins (BMPs) also belong to the 7 has been shown to activate differentiation toward brown
TGF-β superfamily and have been identified as regulators of adipocytes (Tseng et al., 2008). 3T3-F44 2A pre-adipocytes
osteogenesis, and more recently, adipogenesis (James, 2013; treated with BMP-2 showed a decrease in insulin-induced lipid
Moseti et al., 2016), as well as having regulatory roles in accumulation (Skillington et al., 2002), but BMP-7 increased
proliferation, apoptosis, differentiation, and determination of cell the differentiation of 3T3-L1 pre-adipocytes, demonstrating the
fate in adulthood and during embryogenesis (Chen et al., 2004; contradictory roles of BMPs in adipogenesis (Rebbapragada
Wang et al., 2014; Moseti et al., 2016). In contrast to the TGF- et al., 2003; Suenaga et al., 2013). Similarly, BMP-4 regulates
β pathway, BMPs are generally considered as stimulators of the commitment of precursor cells into white adipogenic lineage
both white and brown adipogenesis. Their signal transduction (Gustafson and Smith, 2012). BMP-4 and BMP-7 can also activate
is similar to the TGF-β branch, with differences in the cell the development of beige adipocytes in human precursor cells
surface receptors and types of SMAD proteins involved. Signal (Elsen et al., 2019). Overexpression of BMP-4 in transgenic
transduction begins when BMPs ligands bind to cell surface mice showed a reduction in the size and mass of WAT and
receptors BMPR-1 and BMPR-2. These liganded receptors then induced browning of WAT (Yu et al., 2013). Induced expression
activate (phosphorylate) the R-SMADs (SMAD 1/5/8), which of BMP-4 upregulated the expression of key regulators of
associates with co-SMAD (SMAD4), translocate to the nucleus brown adipose tissue, peroxisome proliferator-activated receptor
and activate target genes (Figure 3). BMPs play different roles gamma coactivator 1-α (PGC-1α) and its target gene, UCP1
FIGURE 3 | A schematic diagram of the TGF-β and BMPs pathway. TGF-β, transforming growth factor-beta; TGF-β-R1,2, transforming growth factor-beta receptor
type 1, 2; co-SMAD, common-SMAD; BMPs, bone morphogenetic proteins; BMP-R1,2, bone morphogenetic protein- type 1 and 2 receptors.

(Qiang et al., 2007; Smith and Kahn, 2016). Likewise, BMP-8b obesity through inhibition of WAT development, and promotion
and BMP-9 are also known to promote brown adipogenesis. of BAT by targeting the key regulating factors of these pathways.
BMP-8b is known to enhance energy dissipation in the body
(Pellegrinelli et al., 2018). It regulates energy metabolism by Wnt Signaling Pathways
increasing BAT thermogenesis both centrally through activation Wnts (Wingless-type MMTV integration site family members)
of AMPK and peripherally through activation of p38 MAPK are secreted glycoproteins that work both in an autocrine
pathway in mature and differentiating brown adipocytes (Whittle and paracrine manner (Moseti et al., 2016) and are post-
et al., 2012; Pradhan et al., 2017). BMP-8b over expression translationally modified by the addition of lipids (Hu et al., 2018).
increases the browning of subcutaneous WAT and enhances its Wnt signaling refers to a group of conserved signal transduction
thermogenic capacity (Pellegrinelli et al., 2018). Whittle et al. pathways consisting of proteins which convey signals through cell
(2012) reported that mice with BMP-8b deletion (BMP-8b−/− ) surface receptors into the cell. These pathways are involved in
exhibited impaired thermogenesis and reduced metabolic rate. cell differentiation and proliferation in adult tissue regeneration,
BMP-9 has been reported to enhance brown adipogenesis in and in embryonic development (Moseti et al., 2016). Abnormal
human adipose-derived stem cells (hASCs). Kuo et al. (2014) activities of various appendages in Wnt signaling pathways causes
showed that a recombinant BMP-9 derivative (MB109) induced the aberrant expansion of adipose tissue (Aamir et al., 2019). Wnt
the thermogenic UCP1 gene mRNA expression and enhanced pathways can be divided into canonical (β-catenin dependent)
brown adipogenesis in hASCs, thus shows anti-obesogenic and non-canonical (β-catenin independent) pathway. MSCs
capacity. These pathways (TGF-β and BMP) are therefore of great are differentiated into osteocytes and myocytes instead of
interest for the discovery of novel chemotherapies in preventing adipocytes upon Wnt/β-catenin signaling pathway activation.
FIGURE 4 | Inhibition of adipogenesis through Wnt/β catenin dependent pathway. Arrows indicate activation and bars indicate inhibition. LRP 5/6, lipoprotein
receptor-related protein 5/6; GSK-3, glycogen synthase kinase-3; APC, adenomatous polyposis coli;TCF/LEF, T-cell factors/lymphoid-enhancing factor; PPARγ,
peroxisome proliferator-activated receptor gamma; C/EBPα, CCCAAT/Enhancer Binding Protein alpha; FAS, fatty acid synthase; aP2, adipocyte fatty acid binding
protein.

Conversely, interruption of Wnt/β-catenin signaling promotes Since the Wnt/β-catenin dependent pathway inhibits
adipogenesis (Figure 4) (Li et al., 2007). Wnt/β-catenin plays adipogenesis and directs the cells toward osteogenesis or
a negative regulatory role in confining the differentiation of myogenesis, its activation constitutes an attractive drug-
adipocytes (Aamir et al., 2019). The signal transduction begins development target to combat obesity and the associated
when Wnt proteins (e.g., wnt 10b) attach to the Frizzled metabolic complications.
receptors and lipoprotein receptor-related protein 5/6 (LRP5/6)
to form a heterotrimeric complex. This complex phosphorylates Hedgehog Signaling Pathway
(activates) Disheveled proteins which disrupt the destruction The Hedgehog (Hh) signaling pathway was first discovered in
complex containing Glycogen synthase kinase-3 (GSK-3)- Drosophila but is now known to be involved in the development
AXIN- adenomatous polyposis coli (APC) (GSK-3-AXIN- of all vertebrates (Liang et al., 2015). The proteins of the Hh
APC), which would otherwise degrade β-catenin. Inhibition family are known as Sonic Hedgehog (SHH), Desert Hedgehog,
of the destruction complex releases and stabilizes β-catenin and Indian Hedgehog (IHH) and participate in the same highly
in the cytoplasm. β-catenin then translocates to the nucleus, conserved Hh signaling pathway (James, 2013) which is an
attaches to T-cell factors/lymphoid-enhancing factor (TCF/LEF) important modulator of stem cell differentiation. Notably, its role
and inhibits adipogenesis through suppression of PPARγ and in the differentiation of MSCs has been demonstrated in several
C/EBPα. Wnt10b is one of the important element of the studies (Fontaine et al., 2008; Plaisant et al., 2009). Hh signaling
Wnt/β-catenin pathway. It has been reported to be responsible initiates when the insoluble and inactive Hh polypeptide
for the anti-adipogenic function of canonical pathway. Wnt10b precursor is converted to a soluble (active) form which makes
is highly expressed in pre-adipocytes, but its expression declines it capable of diffusing across the cell membrane. This modified
promptly after induction of differentiation (Bennett et al., protein then secreted from cell transmembrane proteins named
2002). Its overexpression stabilizes cytoplasmic β-catenin and Dispatched (DISP). After secretion, the Hh polypeptide binds to
blocks adipogenesis in 3T3-L1 pre-adipocytes (Krishnan et al., another cell surface receptor Patched (PTCH) present on nearby
2006; Christodoulides et al., 2009). Similarly, the other two cells. This binding releases another protein called Smoothened
members of the canonical pathway, Wnt6 and Wnt10a have also (SMO), suppressing the PTCH, thus enabling them to activate
been shown to promote osteogenesis and inhibit adipogenesis the glioblastoma gene product (Gli1-3) (James, 2013). Glis are
in St2 and 3T3-L1 pre-adipocytes (Cawthorn et al., 2012). the core transcription factors of this pathway (Figure 5); Gli1 is
Moreover, the Wnt/β-catenin signaling pathway also inhibits used as a reliable marker for the activity of Hh signaling (Tzameli
brown adipogenesis by disrupting the PPARγ and C/EBPα et al., 2004). Hh signaling has inhibitory effects on adipogenesis
induction. Wnt10a and Wnt10b, members of the canonical in murine cells, i.e., KS483, mouse adipose-derived stromal cells,
pathway, are the possible endogenous inhibitors of BAT. Both C3H10T1/2 and calvaria MSC lines (Tang and Lane, 2012). In
Wnt10b and Wnt10a are expressed in pre-adipocytes of BAT mammalian fat, the levels of the components of Hh signaling
but not in differentiated brown adipocytes and their expression respond dynamically to adipogenesis and obesity (Suh et al.,
reduces with the progression of brown adipogenesis (Kang et al., 2006). In mice, subcutaneous fat pad and WAT decrease when the
2005). In addition, Wnt signaling also blocks the thermogenic Hh pathway is activated (Li et al., 2008). Fan et al. (2018) reported
program of BAT by suppressing the thermogenic protein, UCP1 that Hh signaling primarily acts on the later stages of adipocytes
of BAT through repression of PGC-1α (Kang et al., 2005). In vivo differentiation in porcine adipose-derived MSCs. It was revealed
expression of Wnt10b from fatty acid-binding protein 4 (FABP that the expression pattern of Gli1, C/EBPα and PPARγ were
4) promoter had been shown to reduce total body fat by 50% changed on the fourth day of activation of the pathway. Gli1
and provide resistance to WAT accumulation in high-fat diets mRNA and protein expression reached the maximum on the
(Longo et al., 2004). fourth day before gradually decreasing. The mRNA and protein
Members of Wnt family also activate the non-canonical expression of C/EBPα and PPARγ were suppressed significantly
β-catenin independent pathway. As compared to canonical on the fourth day of activation of Hh signaling pathway. Reduced
pathway, less is known about non-canonical β-catenin expression of Gli1, 2, 3 and PTCH promote adipogenesis in MSCs
independent pathway. Members of this pathway include (Fontaine et al., 2008; James, 2013). This signaling pathway is
Wnt 4, 5a/b, 6, 7a/b, and 11 (Ackers and Malgor, 2018). downregulated during the differentiation of human adipocytes
Activation of the non-canonical pathway through Wnt5a is and upon activation, it reduces the expression of C/EBPα
reported to antagonize the canonical pathway, promoting the and thus hinders the accumulation of lipids and adipogenesis
differentiation of pre-adipocytes (Topol et al., 2003). Similarly, (Fontaine et al., 2008; Moseti et al., 2016). Hh signaling pathway
Wnt4 and Wnt5a promote differentiation of adipocytes activation in C3H10T1/2 mouse cell lines was reported to inhibit
(Nishizuka et al., 2008), and Wnt5b together with Wnt5a is PPARγ and C/EBPα expression, blocked the differentiation of
shown to inhibit the Wnt/β-catenin signaling and promotes pre-adipocytes and increased the commitment of C3H10T1/2
adipogenesis by activating PPARγ (Van Tienen et al., 2009). mouse cell lines toward osteogenic lineage (Spinella-Jaegle et al.,
Accordingly, adenoviral overexpression of the related Wnt5b 2001). Activation of Hh gene in a B. mori cell line (BmN)
impaired β-catenin nuclear translocation and enhanced 3T3-L1 inhibited aP2 expression, while knockdown of the Hh gene
cell differentiation (Kanazawa et al., 2005). The non-canonical by RNA interference enhanced aP2 gene expression indicating
pathway, therefore, antagonizes the canonical pathway and the regulatory effect of Hh on aP2. Moreover, the blocking of
promotes adipogenesis. the Hh signaling pathway by an antagonist, cyclopamine, in

FIGURE 5 | Mechanism of action of Hedgehog signaling pathway. Hh, Hedgehog protein; DISP, dispatched protein; PTCH, patched protein; SMO, Smoothened
protein; Gli 1,2,3, glioblastoma gene 1,2,3.
silkworm larvae resulted in increased differentiation and size of of the activity of AMPK (Daval et al., 2006). Stimuli such as
adipocytes. Inhibition of fat formation by Hh signaling pathway exercise, fasting, undernutrition, and exposure to cold result
was retained both in vertebrates and invertebrates (Liang et al., in activation of AMPK in adipose tissue (Daval et al., 2005).
2015). While it has been revealed that activation of Hh signaling For example, in C557Bl/6 mice, AMPK activation increased in
impairs adipogenesis, it is also counterintuitively reported that BAT in response to chronic (>7 days) cold exposure, and in
decrease or blockade of Hh pathway is necessary but not sufficient WAT the activity of α1 AMPK was increased by almost 98%
to trigger adipocyte differentiation (Fontaine et al., 2008; after exposure to cold for more than 15 days (Mulligan et al.,
Fan et al., 2018). 2007). Endogenous stimulators such as high-density lipoproteins
(HDLs), β-adrenergic stimulators, eicosapentaenoic acid and
homocysteine also activate AMPK in BAT of rats and mice,
AMPK AND ITS ACTIVATION IN ADIPOSE and 3T3-L1 adipocytes (Bijland et al., 2013) as does IL 6 in
TISSUE adipose tissue (Daval et al., 2006). Decreased phosphorylation of
AMPK was found in adipose tissue of IL 6 knockout mice after
AMPK is a serine/threonine kinase which is expressed in heavy exercise (Daval et al., 2006). The high concentration of
different kinds of tissues (liver, adipose, skeletal, kidney and adenosine monophosphate (AMP) and low levels of adenosine
hypothalamus) (Kim and Park, 2016) and plays a vital role triphosphate (ATP) resulting from stimuli such as nutrient
in controlling and regulating cell cycle and cellular energy deprivation, ischemia and hypoxia activate AMPK allosterically
homeostasis. AMPK is a fuel-sensing enzyme – it is involved through regulation of an upstream kinase of AMPK (Daval
in sensitivity to, and the homeostasis of, lipids, glucose and et al., 2006; Katwan et al., 2019). In the case of low ATP
insulin (Xu et al., 2012). AMPK activation results in an increase and high AMP levels, the upstream kinase of AMPK; liver
in the body’s cellular energy levels (Kim and Park, 2016). This kinase b1 (LKB1) is activated and phosphorylates AMPK (Bijland
heterotrimeric protein consists of 3 subunits: catalytic subunit et al., 2013). Similarly, the adipokines adiponectin and leptin
α which is comprised of two further subunits α1, α2 and also activate AMPK in adipose tissue (Daval et al., 2006).
regulatory subunits β and γ consisting of two subunits (β1, β2) Overexpression of adipose-specific leptin receptor in WAT of
and three subunits (γ1, γ2, γ3) respectively (Kim et al., 2016; mice leads to an increase in phosphorylation of AMPK Thr172 ,
Hardie, 2018). In adipose tissue, the α1 subunit is considered showing that leptin also activates AMPK in adipose tissue (Wang
to be the most important subunit and accounts for the majority et al., 2005). Orci et al. (2004) reported the phosphorylation of

FIGURE 6 | Activation and functions of AMPK in adipose tissue. Orange arrow and bar indicates functions of ACC and MCoA in the absence of AMPK activation.
Arrows indicate activation and bars indicate inhibition. ACC, acetyl-CoA carboxylase; MCoA, malonyl co-enzyme A; CPT1, carnitine palmitoyltransferase 1;
CaMKK2, calcium/calmodulin-dependent protein kinase kinase 2; LKB1, liver kinase b1; ULK1, Unc-51 Like Autophagy Activating Kinase 1; UCP1, uncoupling
protein 1; PGC-1α, peroxisome proliferator-activated receptor gamma coactivator 1-alpha; SREBP-1c, sterol regulatory element binding protein 1; C/EBPα,
CCCAAT/Enhancer Binding Protein alpha; PPARγ, Peroxisome proliferator-activated receptor gamma.
AMPK in hyperleptinemia white adipocytes. These adipocytes AMPK – its activation correlates with decreased lipid storage
were transformed into “fat burning machines” and it appeared (Bijland et al., 2013). AMPK inhibits de novo synthesis of
that the combustion of fat was due to the leptin-induced cholesterol, triglycerides (TG), and fatty acids (FAs), and
phosphorylation (activation) of AMPK along with increased activates FA uptake and β-oxidation (FAO). It inhibits and
expression PGC-1α and other thermogenic proteins and reduced phosphorylates targets involved in the synthesis of fatty acids
expression of lipogenic proteins. Similarly, activation of AMPK such as FAS, ACC1, and SREBP-1c (Figure 6). SREBP-1c is
by adiponectin in epididymal rat adipocytes is reported by Wu involved in the transcriptional regulation of various lipogenic
et al. (2003). Cellular treatment with adiponectin increased the enzymes, including FAS and ACC1. ACC1 is the predominant
phosphorylation of AMPK at Thr172 and its downstream target form of ACC expressed in lipogenic tissues (Oh et al., 2005;
ACC and resulted in increased glucose uptake. Inhibition of Ridgway and McLeod, 2015). ACC1 converts acetyl-CoA to
AMPK activation by pharmacological agents abrogated glucose malonyl-CoA and catalyzes the rate-limiting step in the synthesis
uptake indicating the activation of AMPK by adiponectin. of FAs (Luo et al., 2011; Jeon, 2016). Malonyl-CoA inhibits
carnitine palmitoyl transferase 1 (CPT1) which is the rate-
limiting enzyme for the transport of fatty acids to mitochondria
Metabolic Functions of AMPK and Role for subsequent oxidation (Bijland et al., 2013). AMPK inhibits
in Adipogenesis the synthesis of cholesterol by phosphorylating and inhibiting
AMPK regulates lipid/glucose homeostasis, mitochondrial HMG-CoA reductase (Jeon, 2016). AMPK also stimulates
biogenesis, autophagy, protein homeostasis, redox equilibrium, mitochondrial biogenesis and β-oxidation through regulation
food intake and insulin signaling (Ceddia, 2013; Jeon, 2016). of PGC-1α activity (Seo et al., 2015). Expression of PGC-1α
Once activated, AMPK directly or indirectly promotes the is related to mitochondrial biogenesis whereas loss of PGC-1α
phosphorylation of downstream targets, including transcription function results in reduced expression of mitochondrial and
and translational factors, metabolic enzymes, epigenetic factors, thermogenic genes in WAT (Kleiner et al., 2012). Wan et al.
growth and proliferation pathways. The overall effect of this (2014) reported that the induction of PGC-1α and the expression
regulation is to reduce the synthesis of cholesterol, fatty acids, of mitochondrial proteins is regulated by AMPK in mouse
ribosomal RNAs (rRNAs) and proteins (Yan et al., 2018). epididymal adipose tissue. AMPK also regulates carbohydrate
Regulation of lipid metabolism is the first known function of metabolism in liver, skeletal muscle and adipose tissue (Kola

et al., 2008; Ceddia, 2013; Jeon, 2016). Skeletal muscle is the of activated AMPK in 3T3-L1 adipocytes. Similarly, Ono and
principal site of insulin-mediated glucose uptake (Koistinen Fujimori (2011) also showed the inhibition of adipogenesis
and Zierath, 2002). In skeletal muscle, AMPK increases glucose through AMPK activation in 3T3-L1 pre-adipocytes. Pollard
uptake through increased glucose transporter type-4 (GLUT-4) et al. (2019) reported that activation of AMPK protects
translocation (Daval et al., 2006; Jeon, 2016). Exercise-stimulated against diet-induced obesity through thermogenesis. Chronic
glucose uptake in skeletal muscle is known to be mediated genetic activation of AMPK resulted in increase of whole-
through the activation of AMPK (Kola et al., 2008). In addition, body energy expenditure which could be due to an increase
AMPK also attenuates glycogen synthesis through inhibition in the consumption rate of oxygen in WAT. AMPK also
of glycogen synthase (GS) and activates glycogenolysis through regulates autophagy (Lee et al., 2018). Several studies have
activation of glycogen phosphorylase (GP) (Jeon, 2016). In demonstrated autophagy in lipophagy, glycophagy, adipose tissue
adipose tissue the potential role of AMPK activation on glucose differentiation and mass regulation (Singh et al., 2009). AMPK
uptake is less clear (Bijland et al., 2013). The majority of the regulates autophagy by phosphorylating two initiating regulators
studies demonstrate that activation of AMPK in white adipocytes of autophagy: a protein kinase complex ULK1 and lipid kinase
inhibits insulin-stimulated glucose uptake (van Dam et al., complex PI3KC3/VPS34 (Kim et al., 2013).
2015a). However, some studies have reported an activating AMPK is also known to have an anti-inflammatory role
effect of AMPK on glucose uptake in adipose tissue. Ye et al. in adipocytes (Mancini et al., 2017) and plays a key role in
(2006) reported enhanced glucose uptake through activation of the inhibition of inflammatory responses (Morita et al., 2018).
AMPK by rosiglitazone in adipose tissue and muscles. Similarly, Inflammation in adipose tissue is known to cause obesity-induced
Attané et al. (2011) and Shen et al. (2014) reported the effect of insulin resistance (Makki et al., 2013). In obesogenic conditions,
AMPK activation on glucose uptake in human adipose tissue the hypertrophied adipocytes and the adipose tissue-resident
and 3T3-L1 adipocytes. Activation of AMPK by apelin in human immune cells increase the levels of circulating proinflammatory
adipose tissue (Attané et al., 2011) and by cinnamon extract in cytokines. Activation of AMPK in adipocytes rapidly suppresses
3T3-L1 adipocytes (Shen et al., 2014) enhanced glucose uptake. the pro-inflammatory pathways (Mancini et al., 2017). Cheng
Inhibition of AMPK by compound-C showed opposite effect et al. (2019) reported that catechin attenuates TNF-α stimulated
which indicates regulation of glucose uptake by AMPK in human inflammation through activation of AMPK/SIRT1 pathway in
adipose tissue and 3T3-L1 adipocytes. 3T3-L1 adipocytes. Similarly, Morita et al. (2018) showed that
In adipose tissue, indirect evidence suggests that activation activation of AMPK reduced the release of MCP-1, which is
of AMPK inhibits differentiation of white pre-adipocytes (Daval known to be one of the most important pro-inflammatory
et al., 2006). AMPK regulates aP2 and induction of C/EBPs adipocytokines. Its over expression in adipose tissue contributes
and PPARγ. AMPK has been shown to inhibit adipogenesis via to infiltration of macrophages and causes chronic low grade
inhibition of the early mitotic clonal expansion (MCE) phase inflammation in adipose tissue (Kamei et al., 2006; Kanda et al.,
accompanied by reduced expression of early and late adipogenic 2006; Morita et al., 2018). Mancini et al. (2017) also showed
factors including FAS, SREBP-1c and aP2 (Habinowski and the anti-inflammatory effects of AMPK in 3T3-L1 adipocytes.
Witters, 2001; Bijland et al., 2013). Vingtdeux et al. (2011) Activation of AMPK inhibited the interleukin 1-β (IL 1-β)
reported the inhibition of adipogenesis by small-molecule stimulated C-X-C motif chemokine 10 (CXCL10) secretion.
activators (RSVA314 and RSVA405) of AMPK via MCE CXCL10 is a proinflammatory cytokine and its upregulation
phase inhibition accompanied by reduced C/EBPβ expression, correlates positively with obesity and type-2 diabetes (Zhang
inhibition of C/EBPα, PPARγ and late adipogenic factors et al., 2014). Activation of AMPK also inhibited the TNF-α
including SREBP-1c, FAS and aP2. Similarly in another study, stimulated IKK/IκB/NFκB signaling (Mancini et al., 2017) which
AMPK activation by A769662 resulted in the reduction of indicates the anti-inflammatory role of AMPK in adipocytes.
lipid droplets and activation of PPARγ, C/EBPα, and early AMPK is obligatory for the proper functioning of BAT as
adipogenic transcription factors such as C/EBPβ and C/EBPδ well (Day et al., 2017). Activation of AMPK increases during the
(Zhou et al., 2009). Likewise, Moreno-Navarrete et al. (2011) differentiation of brown adipocytes, and targeting AMPK with
showed reduced expression of key adipogenic factors such as short interfering RNAs (siRNAs) inhibits the differentiation of
FASN, ACC, PPARγ through activation of AMPK by metformin pre-adipocytes into mature brown adipocytes (Vila-Bedmar et al.,
in human white preadipocytes differentiation. Moreover, it was 2010; Bijland et al., 2013). AMPK is activated in BAT in a situation
observed that increased action of metformin was due to the of chronic cold exposure, providing a thermogenic response
increased expression of organic cation transporter 1 (OCT1 (van Dam et al., 2015a). AMPK is integral to the browning
gene). Cotreatment with cimetidine, an OCT1 gene blocker, of WAT, increasing energy expenditure through thermogenesis
reversed the process resulting in increased adipogenesis and (Chung et al., 2017; Desjardins and Steinberg, 2018). It is vital for
blunted AMPK activity. In addition, He et al. (2013) also reported maintaining the mitochondrial structure, functions, and markers
the inhibition of adipogenesis through activation of AMPK. of mitophagy in BAT. Deletion of AMPK in mice brown and
AMPK activation attenuated the expression of C/EBPα,β and beige adipose tissue causes intolerance to cold exposure and
PPARγ accompanied by decreased expression of SREBP-1c. The reduces thermogenesis in response to β-adrenergic stimulation
phosphorylation of ACC1 and expression of the rate-limiting (Mottillo et al., 2016). These defects were due to impaired
enzyme CPT1 was also increased. These effects were reversed mitophagy which resulted in defective BAT mitochondria, non-
by using AMPK siRNAs, confirming the inhibitory function alcoholic fatty liver disease (NAFLD) and insulin resistance.

AMPK causes mitophagy through phosphorylation of Unc-51 Chen et al. (2012) have shown the activation of AMPK and
like autophagy activating kinase 1 (ULK1) (Sinha et al., 2015; inhibition of 3T3-L1 pre-adipocyte differentiation by CaMKKβ
Mottillo et al., 2016). activation. Activation of CaMKKβ reduced the expression of key
adipogenic factors C/EBPα, PPARγ and SREBP-1 and activated
AMPK Activation by Upstream Kinases in (phosphorylated) AMPK (p-AMPK). Similarly, Lin et al. (2011)
Adipocytes showed the inhibitory effects of CaMMKβ on adipocyte
Under different physiological conditions, the subunits of AMPK differentiation through AMPK activation. Differentiation of pre-
behave differently and are regulated differently. Activation of adipocytes was enhanced in a condition of acute inhibition
AMPK can be achieved by either through upstream kinases or or deletion of CaMKKβ affirming the AMPK activation by
allosterically through AMP (Kim and Park, 2016). The best- CaMKKβ in adipocytes. Likewise, Peng et al. (2012) reported
studied mechanisms of the activation of AMPK are allosteric the activation of AMPK by glucagon through CaMKKβ/AMPK
activation by binding of either AMP or ADP at γ subunit and by pathway. Glucagon enhanced the oxidation of the fatty acid
phosphorylation of the α subunit (Hardie et al., 2012). Conditions and inhibited fatty acid synthesis through phosphorylation of
including hypoxia, exercise, ischemia and hypoglycaemia usually ACC1 at Ser79 and ACC2 through CaMKKβ/AMPK activation
alter the cellular adenine nucleotides levels (suppress ATP in adipocytes. Moreover, it was also observed that fasting
consumption) and subsequently enhance the activity of AMPK led to phosphorylated AMPK and ACC only in CaMKK+/+
(Hardie et al., 2003). The rise in AMP/ADP and decline in the adipocytes but not in CaMKK−/− adipocytes. This demonstrates
levels of ATP cause the activation of AMPK by direct binding of that CaMKKβ/AMPK may be the only pathway through which
ADP or AMP to the γ subunit of AMPK. This binding prevents glucagon regulates lipid metabolism in adipocytes (Peng et al.,
access of phosphatases to Thr172 in the α subunit, and thus 2012). The third upstream kinase of AMPK, TAK1 activates
maintains a high phosphorylation level of AMPK (Jeon, 2016). AMPK-α subunit (Momcilovic et al., 2006; Xiao et al., 2011; Chen
Upstream kinases of AMPK include LKB1, mouse et al., 2013; Wang et al., 2018). It mediates autophagy induced by
protein 25 (MO25) and STE-related adaptor (STRAD), tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)
calcium/calmodulin-dependent protein kinase kinase (CaMKK) in cancerous cells (Herrero-Martin et al., 2009). Although AMPK
and transforming growth factor-β-activated protein kinase is phosphorylated and activated by TAK1 in different tissues and
1 (TAK1) (Oakhill et al., 2012; Desjardins and Steinberg, organs, LKB1 and CaMMKβ are considered the main upstream
2018; Wang et al., 2018). To activate AMPK, LKB1 requires kinases of AMPK in adipocytes (Ceddia, 2013).
two upstream kinases, STRAD and MO25 to join it in a In the case of obesity, AMPK remains inactive due to the
heterotrimeric complex. This complex directly activates AMPK availability of excess nutrients and energy sources, therefore an
by phosphorylating Thr172 of the α subunit. The LKB1/AMPK external stimulus would be needed to activate AMPK. Much
pathway regulates the metabolic check-points of cells and work has been performed to delineate the exogenous activators of
stops proliferation and growth of cells in low ATP conditions. AMPK (Coughlan et al., 2014; Goodman et al., 2014; Kim et al.,
Genetic and biochemical studies in mice, worms and flies 2016) and the debate is still ongoing.
have demonstrated that LKB1 is the major phosphorylating
agent of AMPK (Scott et al., 2008). Shan et al. (2016) reported Exogenous Activators of AMPK
that the presence of LKB1 promoted AMPK activity and its In recent years, much effort has been made to delineate
absence worked oppositely in high-fat diet-induced mice (HFD). the pathways of AMPK and to identify both direct and
Similarly, Hawley et al. (2003) showed that HeLa cells which were indirect activators (Table 1) of AMPK for the development of
unable to express LKB1, upon exposure to external stimuli did new therapies for various disorders including obesity. Many
not elevate AMPK expression. LKB1 activates AMPK in 3T3-L1 pharmacological and natural exogenous activators have been
adipocytes and inhibits adipocyte differentiation (He et al., 2013). reported to activate AMPK either directly independent of
Silencing of LKB1 with siRNAs diminished the activation of upstream kinases or indirectly through upstream kinases.
AMPK in 3T3-L1 adipocytes (He et al., 2013), which pointed to
the activation of AMPK by LKB1 in 3T3-L1 adipocytes. Gormand Direct Exogenous Activators
et al. (2011) have shown that LKB1 is required to maintain the Activators that bind directly to AMPK and activate it without
normal signaling of AMPK in non-stimulated adipocytes. significant changes in ATP:AMP ratio are known as direct
Phosphorylation and activity of AMPK is also promoted by activators. Direct activators induce conformational changes in
other upstream kinases with the lack of expression of LKB1 the AMPK complex, more specifically by interacting with
(Wang et al., 2018). Calcium acts as a trigger for AMPK activation one of the AMPK subunits. 5-Amino-4-imidazolecarboxamide
through calcium/calmodulin-dependent protein kinase kinase- riboside (AICAR) was the first identified direct activator of
2 (also known as CaMKKβ) for phosphorylation of AMPK at AMPK in vitro and in vivo (Fogarty and Hardie, 2010).
Thr172 of the α subunit in some tissues (Desjardins and Steinberg, AICAR has been widely used to evaluate the downstream
2018). Unlike the LKB1 complex, CaMKKβ activates AMPK in effects of activated AMPK in animals (Fogarty and Hardie,
response to an increase in the concentration of cellular Ca2+ 2010). Structurally, AICAR is similar to adenosine and it is
regardless of changes in AMP/ADP/ATP levels (Bijland et al., similarly phosphorylated upon entering the cell (via adenosine
2013). Presence of Ca2+ /CaMKK in adipocytes correspondingly transporters) to AICAR monophosphate (ZMP) by adenosine
regulates the activation of AMPK (Gormand et al., 2011). kinase. ZMP is an analog of adenosine monophosphate (AMP)

TABLE 1 | Some of the direct and indirect activators of AMPK.
Compound Structure AMPK target subunit Activation type References
AICAR γ Direct activation Habinowski and Witters, 2001;

Coughlan et al., 2014
A-769662 β1 Direct activation Desjardins and Steinberg, 2018
Compound 991 (benzimidazole) β1, β2 Direct activation Xiao et al., 2013; Kim et al.,
2016
Salicylate β1 Direct activation Hawley et al., 2012
Compound-2 α1 Direct activation Hunter et al., 2014; Kim et al.,

2016
Metformin α Indirect activation Wang et al., 2019
Troglitazone α Indirect activation Coughlan et al., 2014
Quercetin α, β1 Indirect activation Ahn et al., 2008
Resveratrol α Indirect activation Chen et al., 2011
Curcumin α Indirect activation Ejaz et al., 2009
and similarly activates AMPK allosterically by binding to its Other direct activators of AMPK include A-769662 compound
γ subunit. This causes an increase in Thr172 phosphorylation (Thienopyridone Family), Compound 991 (Benzimidazole
of α subunit of AMPK (Kim et al., 2016) and also inhibits family), and salicylate. A-769662 is a small organic compound
the dephosphorylation of AMPK (Fogarty and Hardie, 2010). which activates AMPK allosterically at Ser108 in the AMPKβ1
Treatment with AICAR has been shown to increase glucose subunit (Kim et al., 2016) and inhibits dephosphorylation of
tolerance, reduce TGs and free fatty acids (FFAs) level of plasma. Thr172 in AMPKα subunit (Sanders et al., 2007; Goodman
AMPK activation by AICAR has been reported to suppress et al., 2014). A-769662 activates AMPK in human primary
the activation of adipogenic transcription factors C/EBPα and subcutaneous adipocytes (Kopietz et al., 2018) and induces
PPARγ, and the enzymes ACC1 and FAS (Habinowski and thermogenesis and browning of inguinal WAT through AMPK
Witters, 2001). Prolonged treatment of AMPK with AICAR signaling (Desjardins and Steinberg, 2018; Wu et al., 2018).
increases BAT- specific protein expression; UCP1 and induces Another direct activator of AMPK is Compound 991 which is
browning of WAT, thus enhances brown adipogenesis (Vila- reported to bind the β unit of AMPK and is more effective (5–10
Bedmar et al., 2010). Although AICAR has these potentially fold) than A-769662 in the allosteric activation and inhibition
useful effects, it also has other AMPK-independent effects of dephosphorylation of AMPK (Xiao et al., 2013; Kim et al.,
which limit its further use (Fogarty and Hardie, 2010). For 2016). Neither A-769662 nor Compound 991 activate AMPK
instance, it acts on other AMP-regulated enzymes such as complexes which contain mutations in the Ser108 of the β subunit
fructose-1,6-bisphosphatase (FBPase) and stimulates muscle of AMPK, suggesting that both A-769662 and Compound 991
glycogen phosphorylase (Longnus et al., 2003; Fogarty and have a similar mechanism for the activation of AMPK (Xiao
Hardie, 2010). In addition, due to short half-life and poor et al., 2013). Likewise, salicylate, a phytochemical obtained from
bioavailability, it is unlikely to be used in the treatment of patients willow bark (Coughlan et al., 2014) and now used very widely
(Coughlan et al., 2014). in the acetylated form (Aspirin) (Goodman et al., 2014), is also

known to activate AMPK. Salicylate is known to directly activate turn increases the release and expression of adiponectin from
AMPK in muscles, liver and WAT (Hawley et al., 2012; van Dam adipocytes (LeBrasseur et al., 2006), which activates AMPK in
et al., 2015b). It binds to the β1 subunit of AMPK and activates skeletal muscle and liver, increases the oxidation of fatty acids
AMPK allosterically, inhibiting the dephosphorylation of Thr172 and uptake of glucose, and decreases the production of hepatic
in the α subunit (Hawley et al., 2012). Beyond these examples, glucose (Kim et al., 2016).
5-(5-hydroxyl-isoxazol-3-yl)-furan-2- phosphonic acid, termed Indirect activation of AMPK by phytochemicals has also been
as Compound-2 (C-2) is the most potent direct activators of reported in numerous studies. Quercetin is one of the most
AMPK. C-2 binds to the AMPKα subunit, causes allosteric abundant flavonoids found in many plants, food and grains, and
activation of AMPK and prevents the dephosphorylation of is known to activate AMPK indirectly (Ahn et al., 2008). Exposure
Thr172 . C-2 mimics AMP’s effects in the activation of AMPK, of 3T3-L1 cells to quercetin resulted in decreased expression of
but unlike AICAR, it does not have any effect on the enzymes positive regulators of adipogenesis, and indeed attenuation of
which use AMP as a substrate (Hunter et al., 2014). C-2 shows adipogenesis. This was due to the phosphorylation of AMPK α
potency twice that of AMP, and 20 times that of A-769662 and β1 subunits, and its downstream substrate ACC (Ahn et al.,
(Kim et al., 2016). 2008). Another indirect activator of AMPK that can be found in
grapes is resveratrol. Resveratrol activates AMPK α indirectly by
Indirect Exogenous Activators increasing AMP:ATP ratio through inhibition of mitochondrial
Studies have shown that modulators which cause calcium ATP production (Fogarty and Hardie, 2010; Hawley et al.,
(Ca2+ ) or AMP accumulation in the body can result in the 2010; Chen et al., 2011). Resveratrol activates AMPK and
activation of AMPK (Kim et al., 2016) without any direct enhances brown adipogenesis (Wang et al., 2015). Treatment
interaction. These modulators are known as indirect activators with resveratrol has been shown to stimulate mitochondrial
of AMPK and may be physiological, pharmacological, or biogenesis, glucose uptake and reduce the accumulation of lipids
natural product activators (Coughlan et al., 2014; Kim et al., in different types of cells (Baur et al., 2006; Zang et al., 2006;
2016). Pharmacological and phytochemical compounds such as Um et al., 2010; Coughlan et al., 2014). Wang et al. (2015)
metformin, troglitazone, quercetin, genistein, epigallocatechin observed enhanced mRNA and protein expression of brown
gallate, resveratrol, berberine, curcumin and the α-lipoic adipocytes-specific markers such as UCP1, PRDM16, PGC-1α
acid act as indirect activators of AMPK (Kim et al., 2016), etc, in inguinal WAT after treatment with resveratrol. It was
activating the kinase by the expenditure of energy because observed that the formation of brown-like adipocyte formation
when ATP is decreased, AMP is increased. Metformin is a in WAT was through activation of AMPK α1. Such brown
biguanide which is found in Galega officinalis (Fogarty and adipocyte formation in WAT was absent in cells lacking AMPK
Hardie, 2010). It upregulates the activity of AMPK, increases α1, which demonstrates the positive role of AMPK in brown
the oxidation of fatty acids, downregulates lipogenic genes, adipogenesis and browning of WAT. In addition, curcumin
increases glucose uptake and decreases the production of derived from Curcuma longa also activates AMPK through its α
glucose. Metformin activates AMPK indirectly, by binding subunit. Exposure of 3T3-L1 adipocytes to curcumin enhanced
and inhibiting the complex I of the mitochondrial respiratory the phosphorylation and activation of AMPK and decreased the
chain, thus increasing the AMP:ATP ratio. It also inhibits the expression of ACC by phosphorylation (Ejaz et al., 2009).
dephosphorylation of AMPK and increases the phosphorylation Unsurprisingly, physiological activators, for instance, exercise
of AMPK through upstream kinase of AMPK, LKB1 (Goodman and calorie restriction, induce the increase in AMP:ATP and
et al., 2014). Metformin mediated activation of AMPK results indirectly activate AMPK. Contraction of muscles both in human
in improved mitochondrial respiration and hyperglycemia in and rodents results in activated AMPK, giving one of the most
obesity. Recently, Wang et al. (2019) have shown the activation compelling examples of molecular effects of exercise (Chen et al.,
of AMPK by metformin in hepatocytes. Metformin mediated 2003; Coughlan et al., 2014). While intracellular energy level is a
AMPK activation promoted mitochondrial fission, improved crucial determinant in the activity of AMPK, it has been reported
mitochondrial respiration and restored the mitochondrial life that reactive oxygen species (ROS) also induced the activation of
cycle. Thiazolidinediones (TZDs) are insulin-sensitizing drugs AMPK without any decrease in ATP level (Quintero et al., 2006;
and consist of rosiglitazone, pioglitazone and troglitazone Wu et al., 2012; Kim et al., 2016).
(Coughlan et al., 2014); these compounds indirectly activate
AMPK and promote phosphorylation of ACC1 in various
types of tissues including adipose, skeletal muscles and liver SUMMARY AND OUTLOOK
(LeBrasseur et al., 2006; Coughlan et al., 2014). Troglitazone
caused phosphorylation and activation of AMPK in adipose Obesity is a common disorder caused by the interaction
tissue just after 15 min of administration (LeBrasseur et al., of environmental, genetic and nutritional factors, and its
2006). Similarly, pioglitazone also caused rapid phosphorylation pervasiveness is accelerating worldwide. Socioeconomic changes,
of AMPK and ACC1 in Swiss 3T3-fibroblast cells (LeBrasseur extensive consumption of calorific foods, and increasingly
et al., 2006). TZDs enhance the accumulation of AMP by sedentary lifestyles are the predominant causative factors for
inhibiting the complex I of the mitochondrial respiratory chain abnormal adipose tissue development and rise in obesity.
and hence activate AMPK indirectly (Brunmair et al., 2004). Abnormalities, both in the development of adipose tissue and the
Moreover, they enhance the expression of PPARγ which in differentiation of pre-adipocytes to mature adipocytes are directly

linked to obesity. Adipose tissue has a strong influence on whole- be investigated in detail, as brown adipogenesis is inversely
body metabolism and therefore is an attractive site for anti- proportional to obesity and associated complications. AMPK,
obesogenic therapies. A cascade of hundreds of transcriptional therefore, can thus be a potential therapeutic target in the
factors and signaling pathways act as either negative or positive prevention and treatment of obesity and we believe that these
modulators of adipose tissue development and adipogenesis. steps could expedite the development of anti-obesogenic drugs
Significant efforts had been made in the past few years to against obesity.
gain insight into the molecular modulation of adipogenesis, but
the investigation into promising targets and identification of
unique regulators of adipogenesis including signaling pathways AUTHOR CONTRIBUTIONS
are still elusive and needed in the fight against obesity. The
heterogeneity of adipose tissue increases the challenges of BA and EW performed the literature search, designed and wrote
determination of the exact role of various signaling intermediates the draft of the manuscript. CS and IF edited the manuscript
and AMPK in different depots. The control of white adipogenesis and contributed to structure and composition. All the authors
accompanied by reduction of lipid contents in mature white reviewed and approved the final version of the manuscript
adipocytes, numeric decrease of adipocytes and controlling before submission.
the abnormal production of cytokines (adipokines) can be an
effective strategy to combat obesity. Moreover, activation of
AMPK in adipose tissue could prove beneficial in attenuating FUNDING
adipose tissue dysfunctionality because AMPK has a crucial
Funding was provided by Taylor’s University Lake Side Campus
role in the regulation of transcriptional factors and pathways
No. 1 Jalan Taylor’s, 47500 Subang Jaya, Malaysia. Taylor’s
related to white/brown adipogenesis and lipid synthesis. AMPK
University Flagship Research Programme (TUFR) Grant Number
activation could also prove beneficial in the prevention of
TUFR/2017/003/05.
various other pathological conditions associated with obesity
such as type-2 diabetes, cancer, chronic inflammation etc.
Obesity is directly linked to chronic inflammation, and chronic ACKNOWLEDGMENTS
inflammation is a risk factor of many diseases. Thus inhibition of
adipose-derived pro-inflammatory cytokines through activation This manuscript is written with the support of Taylor’s
of AMPK could help in the attenuation of metabolic syndrome. University Lake Side Campus No. 1 Jalan Taylor’s, 47500
In addition, the role of AMPK, especially in BAT, must Subang Jaya, Malaysia.
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in adipose-derived stem cells in vitro and in vivo. Calcif. Tissue Int. 79, 169–178. Copyright © 2020 Ahmad, Serpell, Fong and Wong. This is an open-access article
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Transcriptional activation of histone H4 by C/EBPβ during the mitotic clonal original author(s) and the copyright owner(s) are credited and that the original
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doi: 10.1091/mbc.E10-11-0912 use, distribution or reproduction is permitted which does not comply with these terms.

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published: 08 May 2020

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TABLE 1 | Some of the direct and indirect activators of AMPK.

Compound Structure AMPK target subunit Activation type References

AICAR γ Direct activation Habinowski and Witters, 2001;

A-769662 β1 Direct activation Desjardins and Steinberg, 2018

Salicylate β1 Direct activation Hawley et al., 2012

Compound-2 α1 Direct activation Hunter et al., 2014; Kim et al.,

Metformin α Indirect activation Wang et al., 2019

Troglitazone α Indirect activation Coughlan et al., 2014

Quercetin α, β1 Indirect activation Ahn et al., 2008

Resveratrol α Indirect activation Chen et al., 2011

Curcumin α Indirect activation Ejaz et al., 2009

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