Curr Diab Rep (2017) 17: 41
DOI 10.1007/s11892-017-0867-2
PATHOGENESIS OF TYPE 2 DIABETES AND INSULIN RESISTANCE (RM WATANABE, SECTION EDITOR)
The Janus Head of Oxidative Stress in Metabolic Diseases
and During Physical Exercise
Dominik Pesta 1,2 & Michael Roden 1,2,3
Published online: 24 April 2017
# The Author(s) 2017. This article is an open access publication
Abstract
Purpose of Review Oxidative stress describes an imbalance
between production and degradation of reactive oxygen species (ROS), which can damage macromolecules. However,
ROS may also serve as signaling molecules activating cellular
pathways involved in cell proliferation and adaptation. This
review describes alterations in metabolic diseases including
obesity, insulin resistance, and/or diabetes mellitus as well as
responses to acute and chronic physical exercise.
Recent Findings Chronic upregulation of oxidative stress associates with the development of insulin resistance and type 2
diabetes (T2D). While single bouts of exercise can transiently
induce oxidative stress, chronic exercise promotes favorable
oxidative adaptations with improvements in muscle mitochondrial biogenesis and glucose uptake.
Summary Although impaired antioxidant defense fails to
scavenge ROS in metabolic diseases, chronic exercising can
restore this abnormality. The different metabolic effects are
likely due to variability of reactive species and discrepancies
in temporal (acute vs. chronic) and local (subcellular distribution) patterns of production.
This article is part of the Topical Collection on Pathogenesis of Type 2
Diabetes and Insulin Resistance
* Michael Roden
michael.roden@ddz.uni-duesseldorf.de
1
Institute for Clinical Diabetology, German Diabetes Center, Leibniz
Center for Diabetes Research at Heinrich-Heine University
Düsseldorf, Düsseldorf, Germany
2
German Center for Diabetes Research (DZD e.V.),
Munich, Neuherberg, Germany
3
Department of Endocrinology and Diabetology, Medical Faculty,
Heinrich-Heine University, c/o Auf’m Hennekamp 65,
40225 Düsseldorf, Germany
Keywords Reactive oxygen species . Antioxidant capacity .
Obesity . Type 2 diabetes . Exercise
Introduction
Mitochondrial respiration generates reactive oxygen species
(ROS), which are quenched by antioxidant systems. Various
processes such as insulin signaling and upregulation of antioxidants, adaptive protein synthesis, and mitochondrial biogenesis
depend on increased ROS generation under physiological conditions such as exercising. On the other hand, dysregulation of
ROS production and removal, termed oxidative stress, occurs in
numerous human disorders including type 2 diabetes (T2D) and
obesity [1] and has been related to their pathogenesis and complications. This review will address detrimental effects of systemic ROS and ROS originating from skeletal muscle-inducing
oxidative stress and cellular damage in the context of metabolic
diseases but will also explore effects of different exercise training
interventions on oxidative stress in this cohort. This review is
based on a search in biomedical databases (PubMed, Quertle) for
the terms “obese, insulin resistant, type 2 diabetes, ROS, human,
oxidative stress” as well as “obese, insulin resistant, type 2 diabetes, acute, chronic, exercise, ROS, human, oxidative stress”
and mainly focuses on studies published during the last 5 years
but also addresses relevant older studies.
What Is Oxidative Stress?
The term oxidative stress has first been introduced to the biomedical research community in 1985. Oxidative stress has
originally been described as a disturbance in the pro-oxidant–antioxidant balance in favor of the former, potentially
leading to cellular damage [2]. After the discovery of redox
41 Page 2 of 13
Curr Diab Rep (2017) 17: 41
pathways, this definition has been rephrased as “a disruption
of redox signaling and control” [3]. Oxidative stress can also
be defined as a state of temporarily or chronically elevated
ROS production, ROS production is temporarily or chronically elevated, perturbing cellular metabolism and damaging cellular components [4].
ROS are chemical species produced by sequential fourelectron reduction of molecular O2 through the addition of
electrons at metabolically active sites such as the mitochondria
or cytosolic enzymes during their catalytic activity. They comprise of superoxide anion radical (O2·−), hydrogen peroxide
(H2O2), and hydroxyl radical (OH·−), which are chemically
instable and have a strong tendency to react with and damage
biological molecules (Fig. 1). Caloric overload can stimulate
substrate flux to the mitochondria, giving rise to electron
donors (NADH, FADH2) and global electron transport activity with additional electron leakage due to high membrane
potential (ΔΨ). Major sites of mitochondrial net ROS emission include complex I and complex III of the electron transfer
system (ETS) in the inner mitochondrial membrane, the mitochondrial glyceraldehyde-3-phosphate dehydrogenase [5] in
the mitochondrial matrix, as well as the flavoprotein monoamine oxidase in the outer mitochondrial membrane [6, 7].
Other ROS emission sites include the enzymes nicotinamide
adenine dinucleotide phosphate (NADPH) oxidase (Nox) [8]
and nitric oxide synthase (NOS) [9] as well as enzymes that
produce ROS as a byproduct, such as xanthine oxidase (XO)
and lipoxygenase [10]. The process of oxidative protein folding in the endoplasmic reticulum (ER) can also serve as an
important source of ROS and contributes to approximately
Fig. 1 Pathological effects of chronic oxidative stress. Major sources of
reactive oxygen species (ROS) involved in the pathophysiology of insulin
resistance and obesity are shown. The factors responsible for elevated
chronic oxidative stress include inflammatory processes via cytokine
receptors (CR), increased free-fatty acids (FFA) via Toll-like receptor
(TLR), and hyperglycemia, which promote ROS emission from the
NADPH oxidase system (Nox) and nitric oxide (NO) synthase (NOS) as
well as xanthine oxidase (XO) and the mitochondria. Binding of advanced
glycation end products (AGE) to its receptor (RAGE) can further stimulate
Nox-mediated ROS release. Increased mitochondrial oxidation from
nutrient overload and participation of monoamine oxidase (MAO) and
mitochondrial glyceraldehyde-3-phosphate dehydrogenase (mGDPH)
can contribute to excess intracellular ROS production, which can cause
oxidative damage to DNA, proteins, and lipids and also activate the
mitogen-activated protein kinase (MAPK) pathway and C-Jun-Nterminal kinase (JNK), contributing to impaired insulin signaling and
glucose uptake. Elevated intracellular diacylglycerols (DAG) and
ceramides (CER) also interfere with insulin signaling. Endoplasmic
reticulum (ER) stress via the unfolded protein response (UPR) remains
an important source for increased ROS generation. Calcium release from
the ER can enhance cytochrome c release and interfere with electron
transfer within the electron transfer system, thereby further increasing
mitochondrial ROS generation. AKT protein kinase B, CI–CV
mitochondrial complex I–V, ERK extracellular signal-regulated kinase,
GLUT4 glucose transporter 4, IR insulin receptor, IRS insulin receptor
substrate, PI3K phosphoinositide 3-kinase. Arrows denote activation, red
lines indicate inhibition, and words in red represent the most important
outcome of oxidative damage to cellular macromolecules
Curr Diab Rep (2017) 17: 41
25% of overall cellular ROS emission [11]. ER-mediated ROS
emission due to oxidative protein folding increases during
augmented demand in insulin-resistant individuals due to inflammation and higher insulin biosynthesis [12].
Another type of redox signaling molecule, reactive nitrogen species (RNS), is derived from ·NO, a byproduct of Larginine-L-citrulline metabolism catalyzed by NOS isozymes.
·
NO can react with superoxide to form the highly reactive
peroxynitrite (ONOO−). Other RNS include the nitrogen dioxide radical (·NO2) and nitrite (NO2−). RNS may modulate
cell signaling or damage cells by oxidation of biological macromolecules and nitrosylation of other proteins.
In order to detoxify these highly reactive molecules and to
maintain low degrees of oxidative stress, the healthy cell relies
on a large array of antioxidant defense mechanisms. They are
responsible for scavenging and breaking down ROS to less or
nonreactive products and include antioxidant enzymes such as
superoxide dismutase (SOD), catalase (CAT), and glutathione
peroxidase (GPx) as well as endogenous metabolites like bilirubin and uric acid [13]. There are three mammalian isoforms
of SOD: cytosolic Cu/Zn SOD or SOD1, mitochondrial Mndependent SOD or SOD2, and extracellular Cu/Zn SOD or
SOD3, which catalyze detoxification of O2·− to oxygen and
H2O2 [14]. CAT catalyzes the degradation of H2O2 to water
and oxygen, while GPx reduces H2O2 and lipid hydroperoxides to water or corresponding alcohols consuming reduced
glutathione (GSH) [15]. Obviously, both antioxidant defense
and oxidant load affect the redox balance and insufficient
scavenging may be an important cause for oxidative stress.
Methods for Detecting Oxidative Stress
Previous reviews have comprehensively described the
methods for assessment of oxidative stress [16–18]. Briefly,
the challenge of measuring ROS results from their short halflife ranging from nanoseconds to seconds and their overall
very low concentrations. ROS can be either detected directly
or indirectly by measuring molecules that preferably react
with ROS in vivo.
Direct Methods to Assess Oxidative Stress
Highly specific methods involve the trapping of O2·− with spin
trapping reagents such as α-phenyl-N-tert-butyl nitrone (PBN)
that covalently bind and form stable adducts with radicals and
can be detected using quantitative electron spin resonance
(ESR) [16]. Alternatively, rather unspecific spin probes can
be used to detect O2·− in intact tissues, cells, or homogenates.
These probes are oxidized to form stable radicals, which are
then detectable by ESR, a method often regarded as the gold
standard, although methodological limitations, high costs, and
extensive training impede its broad use [19].
Page 3 of 13 41
Alternative approaches to detect ROS in cultured cells, tissues, or isolated mitochondria rely on colorimetic, fluorimetric,
or luminescence-based assays as well as enzyme activity assays.
These assays follow the principle that the radical reacts with a
tracer, which creates a detector compound that releases a photon.
The widely used quantitative lucigenin-enhanced chemiluminescence assay uses lucigenin, a compound reasonably specific for
O2·−[19]. Although this assay is easy to use, it is prone to artifacts
and the validity has been questioned based on O2·− overestimation due to redox cycling of the compound [20]. Lucigenin at
lower concentrations (5 μM) and other compounds such as
coelenterazine, luminol, or methylated-modified cypridina luciferin analog that do not undergo redox cycling are promising
probes for O2·− detection [19].
The widely used Amplex Red assay measures extracellular
H2O2 via the horseradish peroxidase-catalyzed reaction of Nacetyl-3,7-dihydroxyphenoxazine (Amplex Red) with H2O2
in a 1:1 stoichiometry to produce the red-fluorescent oxidation
product, resorufin [21]. Although this assay is highly specific
and sensitive, Amplex Red is light sensitive and thereby prone
to artefactual formation of resorufin at high concentrations of
50 μM [22]. Lowering the concentrations to 10 μM and minimizing light exposure makes the Amplex Red assay an accurate, sensitive, and versatile way for detecting H2O2 emission
from cells, tissues, and cell-free systems.
Indirect Methods to Assess Oxidative Stress
The reaction of thiobarbituric acid (TBA) with the end product of
lipid peroxidation, malondialdehyde (MDA), is among the earliest and most widely used methods for quantitative detection of
lipid peroxides [23]. Although the assay is fast and technically
easy to perform, TBA can also react with other saturated and
unsaturated aldehydes to form unspecific TBA-reactive substances (TBARS) and possibly overestimate MDA levels.
Separation of the aldehyde adducts by high-performance liquid
chromatography (HPLC) has therefore been applied to improve
the sensitivity and accurately quantify MDA levels in tissues and
plasma [24]. Isoprostanes are other important markers of lipid
peroxidation, which can be detected in all body fluids including
urine [25]. F2-isoprostane, the product of peroxidation of arachidonic acid, is considered to be very accurate to quantify in vivo
oxidative stress in plasma or urine [26].
8-Hydroxy-2-deoxyguanosine (8-OHdG), the main product
of DNA oxidation, can easily be assessed in human DNA samples and in urine by HPLC, gas chromatography mass spectrometry (GC/MS), or enzyme-linked immunosorbent assay (ELISA)
[27]. Difficulties arise from the formation of artifacts during isolation and analysis of DNA and from confounding factors such
as smoking [28]. Nevertheless, 8-OHdG is an important marker
for measuring the effect of endogenous oxidative damage to
DNA as a biomarker of oxidative stress.
41 Page 4 of 13
ROS-mediated protein oxidation leads to formation of carbonyl groups (aldehydes and ketones) on protein side chains,
mainly of proline, arginine, lysine, and threonine [29]. These
stable carbonylated proteins can be detected after derivatization
of the carbonyl group with 2,4-dinitrophenylhydrazine (DNPH)
and formation of dinitrophenyl (DNP) hydrazone [29].
Hydrazones are measured spectrophotometrically by ELISA
and Western blotting, the latter yielding semiquantitative results.
Appropriate sample handling and performing the measurement
as quickly as possible help to minimize artifact formation during
sample collection and analysis. Although carbonylated proteins
are induced by various chemical processes and different ROS,
their relatively early formation and stability (hours to days) as
compared to lipid peroxidation products (minutes) make them a
valuable biomarker for oxidative stress [30].
The wide array of methods to detect ROS in different biological matrices offers not only a variety of advantages but
also potential drawbacks. Of note, none of these methods is
generally suitable for every condition. It is therefore recommended to use at least two independent methods to improve
the consistency of experimental observations regarding oxidative stress. The choice should be made considering the sensitivity of the assay in the tested biological specimen. Interest in
more general effects of oxidative stress will lead to the use of
less specific fluorescent probes such as Amplex Red or indirect methods to detect oxidized macromolecules. Interest in
the effects of certain radical species will call for ERS or
lucigenin assays. New methods such as genetically encoded
ROS reporters, nanoparticle delivery systems, and nanotube
ROS probes will likely advance the field by enhanced specificity and sensitivity as well as localization of radical generation [31].
Links Between Oxidative Stress and Metabolic
Diseases
John Baynes was one of the first to present the hypothesis that
oxidative stress could be an important mechanism contributing to
the pathogenesis of diabetes, arguing that diabetes-related complications are associated with oxidative damage of proteins and
lipids [32]. Subsequent studies supported this hypothesis in that
increased systemic and skeletal muscle ROS production may
relate to the development of several metabolic abnormalities including obesity and T2D [33, 34].
Systemic Oxidative Stress in Obesity and Insulin
Resistance
Several studies reported mainly elevated oxidative stress in metabolic disorders in humans (Table 1). In obese nondiabetic
humans, different parameters of fat accumulation correlate with
systemic oxidative stress [35, 36]. Similarly, oxidative DNA
Curr Diab Rep (2017) 17: 41
damage is elevated in individuals with prediabetes, defined by
impaired fasting blood glucose >6 mmol/l but <7 mmol/l [37].
While antioxidant capacity (SOD, CAT) is also upregulated in
young obese individuals, elderly obese persons and individuals
with metabolic syndrome show decreased antioxidant capacity in
the face of increased systemic lipid peroxidation and protein
carbonyls in parallel with altered lipoprotein metabolism and
decreased antioxidant capacity, and both oxidative stress and
antioxidant capacity (SOD, CAT) are upregulated in young obese
individuals [38, 39]. Thus, systemic oxidative stress is present in
obese and insulin-resistant individuals, which further rises with
aging and progression of metabolic abnormalities due to inadequate upregulation of antioxidant defense.
Tissue-Specific Oxidative Stress in Obesity and Insulin
Resistance
In a healthy cohort covering a wide range of body masses (body
mass index (BMI) 18 to 37 kg/m2), vascular oxidative stress, and
expression of Nox-p47phox, a Nox accessory protein involved in
O2·− production rises with the degree of adiposity without alteration of xanthine oxidase activity [40] (Table 1). The overweight/
obese group features elevated protein carbonyls despite higher
vascular CAT and SOD expression, suggesting compensation for
increased oxidative stress. Notably, systemic plasma markers of
oxidative stress and antioxidants are not different between normal weight and overweight/obese individuals in this study [40].
Obese insulin-resistant individuals exhibit twofold higher muscle
mitochondrial H2O2 emission than healthy controls, possibly related to lower protein abundance of complex I subunits as well as
enzymes responsible for the oxidation of fatty acids and
branched-chain amino acids [41]. A similar increase in muscle
mitochondrial H2O2 emission is paralleled by a 50% reduction in
the GSH/GSSG ratio in muscles of obese insulin-resistant individuals [42]. Of note, others report unchanged or even decreased
rates of H2O2 emission in obese insulin-resistant individuals, but
these results are difficult to interpret in the absence of data on
maximal ROS production rates [43]. Nevertheless, the above
observations suggest that increased fat mass and insulin resistance may favor ROS production from vascular endothelium
and skeletal muscle with insufficient tissue-specific compensation by antioxidant systems, leading to increased systemic oxidative stress independent of age and hyperglycemia.
Associations of Fat Mass and Insulin Resistance With
Oxidative Stress
Obese children with greater insulin resistance as assessed by
the homeostasis model assessment of insulin resistance
(HOMA-IR) also present with a higher degree of oxidative
stress [44]. In addition to insulin resistance, circulating triglycerides and high-sensitivity C-reactive protein (hsCRP) may be
involved in the relationship between adiposity and oxidative
Curr Diab Rep (2017) 17: 41
Page 5 of 13 41
Table 1 Effects of ROS on the systemic and cellular environment in human obesity, insulin resistance, and/or type 2 diabetes (studies are listed in
chronological order)
Author
Population
Biological matrix
Pro-oxidants
Antioxidants
Shin et al. [49]
T2D (n = 41)
HC (n = 33)
T2D (n = 25)
HC (n = 20)
T2D (n = 7)
oHC (n = 5)
yHC (n = 9)
HC (n = 140)
Serum
↑8-OHdG
n.a.
Urine
↑8-OHdG
n.a.
Vastus lateralis muscle
n.a.
Plasma and urine
Erythrocytes and plasma
↑TBARS, F2-isoprostane
associated with body fatness
↑TBARS
↓HSP72; ↓HO-1 T2D
↔HSP72 and HO-1
vs. yHC
n.a.
↓GPx, CAT, GSH
Endothelial cells
↑Nox
↑CAT, SOD
Erythrocytes and plasma
↑MDA in T2D
↑DNA damage in T2D and IGR
Vastus lateralis muscle
↑H2O2 emission
↓SOD in IGR vs. HC
↓SOD and TAC in T2D
vs. HC
↓GSH/GSSG
Vastus lateralis muscle
↓H2O2 emission in OB-IR
↔in T2D and HC
↑H2O2/ATP ratio in T2D
n.a.
Plasma
OxLDL and F2-isoprostanes
associated with IR
↑H2O2 emission
↔mito respiration
↓complex I subunits
↑8-OHdG in T2D and PRE vs. HC
n.a.
Erythrocytes and plasma
↑hydro peroxides in OB
↑protein carbonyls in OB
T2D (n = 10)
OB (n = 10)
HC (n = 10)
yOB (n = 40)
Vastus lateralis muscle
↑protein carbonyls in T2D
↑ROS in FTO overexpressing cells
↓Total antioxidant
capacity in OB
↑SOD, CAT in yOB
↓SOD, CAT in oOB
↓GPx in OB
↓SOD2 in T2D
Plasma
MS (n = 14)
HC (n = 13)
African American
(n = 82)
White American (n = 76)
T2D (n = 68)
Plasma and serum
Lipid peroxidation and protein carbonyls
correlate with IR
↑TBARS
Kanauchi et al. [48]
Bruce et al. [54]
Furukawa et al. [35]
Dave and Kalia [52]
Silver et al. [40]
Song et al. [51]
Anderson et al. [42]
Abdul-Ghani et al. [43]
Park et al. [47]
T2D (n = 50)
HC (n = 50)
OB/OW (n = 42)
HC (n = 39)
T2D (n = 113)
IGR (n = 78)
HC (n = 92)
Male OB-IR (n = 3)
HC (n = 5)
T2D (n = 10)
OB-IR without T2D
(n = 10)
HC (n = 10)
HC (n = 5115)
Lefort et al. [41]
OB-IR (n = 14)
HC (n = 20)
Vastus lateralis muscle
Al-Aubaidy and Jelinek [37]
T2D (n = 35)
PRE (n = 8)
HC (n = 119)
yOB (n = 45)
oOB (n = 40)
yHC (n = 65)
oHC (n = 55)
Serum
Karaouzene et al. [39]
Bravard et al. [53]
Codoner-Franch et al. [44]
Yokota et al. [38]
Warolin et al. [36]
Ohara et al. [55]
Kant et al. [50]
T2D or prediabetes
(n = 43)
HC (n = 37)
Urine
Plasma
Urine
F2-isoprostane positively correlated with
body fatness
↔between groups
↑d-ROMs associated with daily glucose
variability
↑8-OHdG, S-cdA, and 8-iso-PGF2α
n.a.
n.a.
n.a.
↓total thiols
↓SOD
n.a.
n.a.
n.a.
8-OHdG 8-hydroxy-2′-deoxyguanosine, 8-iso-PGF2α 8-iso-prostaglandin F2α, ATP adenosine triphosphate, CAT catalase, CS citrate synthase, DNA
deoxyribonucleic acid, d-ROMs diacron-reactive oxygen metabolites, FTO fat mass and obesity associated, GPx glutathione peroxidase, GSH glutathione, GSSG glutathione disulfide, HC healthy controls, HSP heat shock protein, IGR impaired glucose regulation, IR insulin resistant, MDA
malondialdehyde, MS metabolic syndrome, n.a. not assessed, Nox nitric oxide synthase, o old, OB obese individuals, OW overweight individuals,
ROS reactive oxygen species, SOD superoxide dismutase, S-cdA (5′S)-8,5′-cyclo-2′-deoxyadenosines, T2D individuals with type 2 diabetes, TBARS
thiobarbituric acid reactive substances, y young
stress as adjustment for these parameters reduces the association. Indeed, elevated hsCRP strongly associates with oxidative stress independent of BMI and insulin resistance [45, 46].
Furthermore, plasma adiponectin levels are negatively
associated not only with BMI and waist circumference but
also with markers of systemic oxidative stress. In a
population-based observational study of 5115 individuals,
the positive relationship of levels of oxidative stress markers
41 Page 6 of 13
with HOMA-IR disappears for F2-isoprostane after adjustment for adiposity but remains for oxidized low-density lipoprotein (ox-LDL) [47].
Taken together, these results suggest that fat accumulation,
insulin resistance, and deranged lipoprotein metabolism can
contribute to increased oxidative stress independent of hyperglycemia. However, these studies do not allow to draw conclusions on causal relationships.
Systemic Oxidative Stress in T2D
T2D patients show elevated markers of oxidative DNA damage in plasma and urine [48, 49] (Table 1). Of note, oxidative
DNA damage and lipid peroxidation are already present in
newly-diagnosed T2D and even in prediabetic individuals,
defined as fasting blood glucose between 5.5 and 7 mmol/l
and hemoglobin HA1c (HbA1c) levels of 5.7 to 6.4% [50].
While individuals with impaired glucose regulation (impaired
fasting glucose and impaired glucose tolerance) show similar
levels of oxidative stress, but slightly reduced erythrocyte
SOD activity compared to glucose-tolerant people, levels of
plasma lipid peroxides, and DNA damage are elevated and
total antioxidant capacity, GPx, GSH, and SOD activity are
decreased in T2D [51, 52].
Tissue-Specific Oxidative Stress in T2D
Oxidative tissue damage measured as increased protein carbonyls is present in the skeletal muscle of T2D individuals in
parallel with decreased SOD activity [53] (Table 1). Also,
expression of heat shock protein (HSP)72 and heme oxygenase (HO)-1, genes responsible for antioxidant defense mechanisms, is markedly lower in the skeletal muscle of T2D [54]
(Table 1). In T2D, muscle expression of antioxidant genes
further correlates with muscle oxidative capacity and insulinstimulated glucose disposal [54, 55].
Collectively, these results suggest that even moderate increases in blood glucose impair antioxidant defense, which
leads to oxidative damage with possible deterioration of skeletal muscle function in overt diabetes.
Associations of Hyperglycemia and Diabetes With
Oxidative Stress
The reduction in antioxidant defense associates negatively
with whole-body insulin sensitivity. Markers of oxidative
DNA damage correlate with BMI, hyperglycemia, and βcell dysfunction and progressively increase from prediabetic
(5.5 and 7 mmol/l) to diabetic conditions [37, 51]. On the
other hand, the positive correlation of ox-LDL and lipid peroxidation and the negative correlation of total antioxidant
levels and SOD activity with insulin resistance were found
to be independent of obesity in one study [47]. In T2D, muscle
Curr Diab Rep (2017) 17: 41
expression of antioxidant genes further correlates with muscle
oxidative capacity and insulin-stimulated glucose disposal
[54, 55]. Finally, oxidative stress is further associated with
daily and day-to-day glucose variability [54, 55]. Taken together, glycemic control is an important driving force for further accelerating oxidative stress and impairment of antioxidant defense.
Results From Diet Intervention Studies
Ingestion of a high-fat diet increases mitochondrial H2O2 emission and induces insulin resistance in healthy males [42], but
not in obese insulin-resistant women [56•]. High-fat diet-induced oxidative stress in skeletal muscle also associates with
reduced expression of muscle mitochondrial oxidative phosphorylation genes [42, 57]. However, comparison of a highcarbohydrate and a high-fat meal reveals that only the highcarbohydrate meal decreases total antioxidant capacity and
muscle SOD, supporting the view of the deleterious role of
carbohydrates and glycemia for oxidative stress [58].
On the other hand, diet-induced weight loss can decrease
oxidative stress by improving antioxidant status independent
of physical activity [59]. In obese women, body weight reduction not only improves insulin resistance, oxidative stress, and
inflammation but also activities of antioxidant enzymes including GSH and CAT [60, 61]. Similarly, a 2-month calorie
restriction by 20% resulting in 8% weight loss leads to reduction in dyslipidemia as well as markers of oxidative stress and
inflammation along with improved antioxidant defense [62].
Taken together, intervention studies suggest that caloric intake
and body weight changes dynamically affect oxidative stress
but do not allow to identify whether oxidative stress contributes to the weight-dependent alterations in metabolism and
insulin resistance.
Cellular Mechanisms Contributing to ROS Production
in Human Metabolic Diseases
At least 0.2–2% of the oxygen consumed during mitochondrial respiration contributes to the generation of free radicals [63,
64] (Fig. 1). T2D patients exhibit slightly lower flux through
muscle ATP synthesis as well as in muscle expression of genes
involved in mitochondrial function and oxidative metabolism
[65–67]. Incomplete mitochondrial catabolism of long-chain
fatty acyl-CoA has been further associated with elevated ROS
production and impaired glutathione antioxidant system [68].
Intracellular accumulation of lipid metabolism intermediates
(diacylglycerols, ceramides) further impairs insulin signaling
[69••]. However, the data on muscle mitochondrial function in
insulin-resistant (IR) and T2D humans are not fully consistent
in that some features of mitochondrial function are comparable between T2D and age-matched glucose-tolerant
Curr Diab Rep (2017) 17: 41
individuals when respiratory rates are normalized to mitochondrial DNA or citrate synthase activity.
Adipose tissue may serve as an important source of ROS:
nondiabetic obese KKay mice exhibit increased lipid peroxidation and H2O2 production in white adipose tissue. Elevated
circulating fatty acids could contribute to oxidative stress via
NADPH oxidase activation in white adipose tissue [35].
Obesity also relates closely to ER stress [70], which, in turn,
associates with oxidative stress [71]. ER-mediated ROS production is increased in both obese insulin-resistant nondiabetic
persons and T2D patients [72, 73]. In the context of ER stress,
unfolded protein response (UPR) leads to calcium ion leakage
from ER, which interferes with electron transfer in the ETS
[74] and the subsequent cytochrome c release from mitochondria can induce mitochondrial ROS production [75] (Fig. 1).
Finally, advanced glycation end products (AGEs) increase
under conditions of hyperglycemia [76]. Binding of AGEs to
their respective receptors (RAGE) stimulates Nox, which also
generates intracellular ROS [77]. (Fig. 1).
Effects of Acute and Chronic Exercise on Oxidative
Stress in Metabolic Diseases
Since the 1970s, it is known that 1 h of moderate endurance
exercise can increase lipid peroxidation in humans [78].
Although the biological meaning was unknown, these results
created a lot of interest over the following years about the role
ROS plays during exercise. While acute exercise may induce a
temporary state of oxidative stress, chronic physical activity
promotes favorable oxidative adaptations [79]. Various modalities of regular exercising (endurance, resistance, or combined training) generally improve systemic markers of oxidative stress and antioxidant capacity in healthy individuals [80,
81], but its impact on oxidative stress in metabolic diseases is
less clear.
Acute Exercise and Oxidative Stress in Metabolic Diseases
An acute bout of exhaustive aerobic exercise results in greater
ROS production in obese than in nonobese individuals [82]
(Table 2). Both intensive aerobic and resistance exercise sessions lead to excessive lipid peroxidation in obese men and
women [83, 84]. However, low-intensity exercise such as
walking decreases lipid peroxidation in individuals with
T2D, suggesting that mild exercising is able to reduce systemic oxidative stress in T2D [85]. Of note, acute high-intensity
exercise-induced oxidative stress associates with increased insulin sensitivity in obese individuals [86]. Experimental studies in myocytes mimicking acute and chronic oxidative stress
support this concept [87]. During acute and oxidative stress,
the mitogen-activated protein kinase (MAPK) phosphatase
MKP7 relocates from the nucleus to the cytoplasm, where it
Page 7 of 13 41
dephosphorylates JNK in the cytoplasm, resulting in increased
insulin sensitivity through insulin receptor substrate (IRS)-1.
These results suggest that oxidative stress in response to exercise is exacerbated in individuals with metabolic diseases
but may also serve as an important signal improving insulin
signaling and mitochondrial biogenesis.
Chronic Exercising and Systemic Oxidative Stress
in Obesity and Insulin Resistance
In overweight-to-obese women, 12 weeks of aerobic exercise
training plus caloric restriction lowers systemic lipid peroxidation [88] (Table 2). In a similar 12-week intervention study,
combined aerobic exercise and hypocaloric diet is more effective to decrease oxidative stress and improve serum antioxidant capacity than hypocaloric diet alone [89]. On the other
hand, obese women feature reduction in markers of lipid peroxidation upon aerobic exercising plus caloric restriction or
caloric restriction alone, but only in those individuals increasing their maximal oxygen uptake upon exercising alone [90].
Both regular aerobic and resistance training alone can improve
oxidative stress and antioxidant defense in overweight individuals [81, 91]. Five weeks of combined resistance and aerobic training improve both oxidative stress and insulin resistance in insulin-resistant humans [92]. However, the combined exercise training can lead to higher oxidative lipid damage in obese individuals [92], suggesting that caution may be
required when recommending combined training in persons
with metabolic diseases.
Chronic Exercising and Tissue-Specific Oxidative Stress
in Obesity and Insulin Resistance
As little as 2 weeks of immobilization diminishes muscle ATP
production and increases muscle H2O2 emission without effects on antioxidant proteins, while a subsequent 6-week period
of aerobic training not only restores ATP production and H2O2
emission to baseline levels but also increases SOD and CAT
[93•] (Table 2). Likewise, a 12-week aerobic exercise training
period reverses muscle mitochondrial alterations, diminishes
cellular oxidative damage, and increases CAT activity in obese
insulin-resistant women despite minimal weight loss [56•].
Chronic Exercising and Systemic Oxidative Stress in T2D
Chronic aerobic exercise training can reduce oxidative damage
to proteins, lipids, and DNA as well as improve systemic antioxidant status in obese and T2D individuals [90, 94, 95]
(Table 2). A 12-month supervised exercise training period
consisting of aerobic, resistance, and flexibility training reduces
some features of oxidative stress independent of changes in
body weight [96••], but not systemic lipid peroxidation [97].
In obese individuals with impaired glucose tolerance, 12-weeks
Effects of acute and chronic exercise interventions on ROS and energy metabolism in humans with metabolic diseases (studies are listed in chronological order)
Author
Acute exercise
Vincent et al. [83]
Vincent et al. [84]
Roh et al. [82]
Haxhi et al. [85]
Parker et al. [86]
Chronic exercise
Kasimay et al. [99]
Gutierrez-Lopez et al. [89]
Brinkmann et al. [101]
de Oliveira et al. [97]
McNeilly et al. [98]
Krause et al. [102]
Vinetti et al. [96••]
Pittaluga et al. [94]
Medeiros et al. [92]
Gram et al. [93•]
Bianchi et al. [88]
Farinha et al. [91]
Konopka et al. [56•]
Intervention
(duration; mode; frequency)
Biological matrix
Pro-oxidants
Antioxidants
OB (n = 14)
HC (n = 14)
OW (n = 24)
HC (n = 8)
OB (n = 12)
HC (n = 12)
T2D (n = 9)
OB (n = 11)
AE, RE
Plasma
↑lipid peroxides in OB after AE and RE
↑antioxidant capacity in HC after RE
AE
Plasma
↑lipid peroxides in OW after AE
↔thiols
AE
Plasma
↑ROS in OB
↑SOD after AE
AE
HIIT
Urine
Vastus lateralis muscle and plasma
↓F2-isprostanes
↑JNK/MAPK
n.a.
↑insulin-stimulated SOD
OB-IGT (n = 14)
OB (n = 32)
HC (n = 16)
T2D (n = 15)
OB (n = 12)
T2D (n = 43)
12 weeks; CR + AE; 3/week
12 weeks; CR, CR + AE; 3/week
Plasma
Plasma
↓lipid peroxides
↓lipid peroxides, protein carbonyls
↑SG
n.a.
12 weeks; AE; 3/week
AX before and after
12 weeks; AE, RE, CT, NT; 3/week
Plasma, erythrocytes
Plasma
↓F2-isprostanes with AE, ↔with AX in
T2D and OB
↔lipid peroxides
12 weeks; mild AE; 5/week
16 weeks; AEL, AEM; 3/week
Serum
Plasma and vastus lateralis muscle
↓lipid peroxides
↑protein carbonyls in T2D; ↑NO, NOS in HC
↑peroxiredoxin oxidation in T2D
with AX after AE
↑CAT, SOD, NO, SG with AE;
↔with RE; ↑SG with CT
↔SOD
↑CAT in T2D with AEM
52 weeks; RE, AE, FL; 3/week
12 weeks; AE, 3/week
Plasma and PBMC
Plasma
↓lipid peroxides
↓MDA, DNA damage in T2D
n.a.
↑GSH, AA
5 weeks; CT1; 5/week
9 weeks; CT2; 3/week
2 weeks IM followed by 6 weeks; AE; 3/week
Plasma
↓GPx
12 weeks; CR + AE; 3/week
12 weeks; AE; 3/week
Plasma
Plasma and serum
12 weeks; AE; 3/week
Vastus lateralis muscle
↓protein carbonyls CT1
↑protein carbonyls CT2
↑H2O2 emission, ↓ATP generation after IM,
reversed by AE
↓hydroperoxides
↓lipid peroxides
↓protein carbonyls
↓H2O2 emission and DNA damage
12 weeks; AE; 3/week
52 weeks; CR, AE, CR + AE, NT; 3/week
2 weeks; IW, CW, NT; 5/week
10 weeks; RE; 3/week
Plasma
Plasma
Plasma, urine
Plasma
↓protein carbonyls
↓F2-isprostanes in CR and CR + AE, ↔in AE
↔F2-isprostanes
↓MDA
OB-IGT (n = 11)
T2D (n = 13)
HC (n = 12)
T2D (n = 20)
T2D (n = 12)
HC (n = 12)
OB (n = 25)
yHC (n = 17)
eHC (n = 15)
OOW (n = 50)
MSW (n = 23)
OW (n = 25)
HC (n = 14)
T2D (n = 31)
OOW (n = 439)
T2D (n = 14)
HEW (n = 25)
Vastus lateralis muscle
↑CAT, SOD with AE
n.a.
↑total thiols
↑CAT
↑sialic acid
n.a.
n.a.
↑H2O2 scavenging
AA ascorbic acid, AE aerobic exercise, AEL aerobic exercise low, AEM aerobic exercise moderate, AX acute exercise, CAT catalase, CR calorie restriction, CT concurrent training, CW continuous walking,
GPx glutathione peroxidase, GSH glutathione, HC healthy controls, HEW hypertensive elderly women, IGR impaired glucose regulation, IGT impaired glucose tolerance, IM immobilization, IW
intermittent walking, MDA malondialdehyde, MSW women with metabolic syndrome, n.a. not assessed, NO nitric oxide, NOS nitric oxide synthase, NT no treatment, OB obese, OOW overweight/obese
women, PBMC peripheral blood mononuclear cells, RE resistance exercise, SG sulfhydryl groups, SOD superoxide dismutase
Curr Diab Rep (2017) 17: 41
Dincer et al. [95]
Duggan et al. [90]
Karstoft et al. [103]
Dantas et al. [81]
Population
41 Page 8 of 13
Table 2
Curr Diab Rep (2017) 17: 41
of mild aerobic training decreases body mass, percent body fat,
and systemic lipid peroxidation and improves insulin sensitivity without affecting SOD [98]. As in insulin-resistant persons,
aerobic training in combination with caloric restriction reduces
oxidative stress and improves antioxidant capacity in obese
glucose-intolerant persons [99]. The precise interaction between these two interventions remains to be established, as
exercise training seems to provide no additional benefit when
used in combination with caloric restriction [100]. Although
individuals with T2D show elevated oxidative stress after
maximal-intensity exercise compared to healthy individuals,
12 weeks of preconditioning with regular aerobic training
markedly diminished oxidative stress in response to an acute
bout of exercise in individuals with T2D [101].
Chronic Exercising and Tissue-Specific Oxidative Stress
in T2D
Only a few studies have addressed the effect of exercise on
muscle oxidative stress in T2D (Table 2). A 16-week period of
unsupervised aerobic exercise training at moderate intensity is
more effective to attenuate muscle oxidative protein damage
and to increase CAT activity in obese T2D patients than exercise training at low intensity [102]. These effects occurred
without changes in insulin sensitivity and body composition.
On the other hand, reductions in oxidative stress related to
improvements in insulin sensitivity [96••]. In another study,
only interval walking training but not continuous walking
training improved glycemic control without any effect on oxidative stress in T2D patients [103]. These findings indicate
that the training response of oxidative stress and antioxidant
defense is dependent on intensity, at least at lower levels of
exercising, but again does not necessarily relate to insulin
sensitivity or glycemic control.
Although the effect of exercise on oxidative stress is somewhat mixed and dependent on the metabolic disease, individuals with metabolic diseases may experience an exacerbation
of oxidative stress following exercise when compared to
healthy individuals. However, this increased oxidative stress
may act as a preconditioning and induce upregulation in antioxidant defenses, which leads to diminished levels of oxidative stress when experiencing subsequent pro-oxidant environments [104].
Cellular Mechanisms Contributing to Exercise-Mediated
ROS Production in Human Metabolic Diseases
In contrast to chronic oxidative stress in metabolic diseases,
exercise-induced ROS occurs transiently, mostly limited to the
duration of the exercise session. Thus, the pattern of ROS
production follows the concept of hormesis, i.e., favorable
biological effects at low exposures and opposite effects at
higher doses. Adaption in response to exercise-induced
Page 9 of 13 41
oxidative stress renders the cell less vulnerable to successive
perturbations [105]. ROS produced during higher-intensity
exercise possibly promotes glucose uptake and improves glycemic control. In the mouse, repetitive contractions of extensor digitorum longus muscle stimulate glucose uptake by
300%. Contraction-stimulated, but not basal glucose uptake
decreases by 50% after the addition of the antioxidant N-acetyl cysteine (NAC) [106]. Treatment with ebselen, an antioxidant reducing H2O2, similarly reduces glucose uptake, indicating a role of H2O2 generation [107]. Interestingly, these
results could not be confirmed during in situ studies of NAC
infusion in rats [108] and humans [109], possibly due to different NAC concentrations. Alternatively, ROS may only influence contraction-mediated glucose uptake during higherintensity exercises with greatest ROS production [110]. Of
note, treatment with the ROS scavengers and vitamins C and
E surprisingly prevents exercise-induced increases in insulin
sensitivity in healthy individuals, suggesting a beneficial role
of ROS under exercise conditions [111].
Exercise-induced ROS also stimulates mitochondrial biogenesis and can improve mitochondrial function. Peroxisome
proliferator-activated receptor gamma coactivator 1 alpha
(PGC-1α) is a key regulatory factor mediating mitochondrial
biogenesis [112], a process which also involves activation of
AMP-activated protein kinase (AMPK) [113] (Fig. 1). Indeed,
H2O2 treatment of C2C12 cells results in enhanced AMPK
activation and PGC-1α promotor activity, both of which are
blocked by NAC [114].
In summary, ROS generated during exercise likely mediates favorable adaptations including improvements in glucose
uptake and mitochondrial biogenesis.
Conclusions
Detecting oxidative stress remains challenging, although novel developments may improve the diagnostic efficacy of ROS
measurement. Nevertheless, reliable and easy-to-use biomarkers of redox homeostasis will be required to assess oxidative stress in clinical studies.
Numerous studies support the concept of a compromised
balance between ROS generation and the antioxidant defense
network in obesity and insulin-resistant states. The resulting
chronic oxidative stress contributes to insulin resistance.
Exercise training can ameliorate these effects and result in
adaptive responses and improved endogenous antioxidant capacity in individuals with metabolic diseases. These changes
can occur independent of relevant weight loss. Although acute
exercise might induce a short-term pro-oxidative environment,
regular exercise, regardless of the modality, improves cellular
antioxidant capacity in obese, T2D, and IR individuals.
Chronic exercise training can reduce oxidative stress, but this
41 Page 10 of 13
reduction is not necessarily related to improved insulin sensitivity and/or glycemic control.
However, the precise localization and origin of ROS in
different pathological and physiological conditions and the
effect of specific ROS on specific signaling pathways remain
unclear. Specifically, comprehensive human studies exploring
triggers for ROS generation during acute and chronic exercise
and the impact of ROS on important cellular signaling pathways in the context of exercise adaptation and development of
T2D are still lacking.
Acknowledgments The work of the authors is supported in part by the
Ministry of Science and Research of the State of North Rhine-Westphalia
(MIWF NRW) and the German Federal Ministry of Health (BMG) as
well as by a grant of the Federal Ministry for Research (BMBF) to the
German Center for Diabetes Research (DZD e.V.; DZD Grant 2012) and
by grants from the Helmholtz portfolio theme: Metabolic Dysfunction
and Common Disease and the Helmholtz Alliance to Universities:
Imaging and Curing Environmental Metabolic Diseases (ICEMED), the
German Research Foundation (DFG, SFB 1116), German Diabetes
Association (DDG), the Schmutzler-Stiftung and by the Austrian
Science Fund (FWF), project no. J-3267.
Curr Diab Rep (2017) 17: 41
3.
4.
5.
6.
7.
8.
9.
10.
11.
Compliance with Ethical Standards
12.
Conflict of Interest Dominik Pesta declares that he has no conflict of
interest.
Michael Roden reports personal fees from Astra Zeneca, Eli Lilly, GI
Dynamics, Merck, and Novo Nordis; grants and personal fees from
Boehringer Ingelheim; grants from Novartis Pharma; and grants and personal fees from Sanofi.
13.
14.
Human and Animal Rights and Informed Consent This article does
not contain any studies with humans or animals performed by any of the
authors.
Open Access This article is distributed under the terms of the Creative
Commons Attribution 4.0 International License (http://
creativecommons.org/licenses/by/4.0/), which permits unrestricted use,
distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the
Creative Commons license, and indicate if changes were made.
15.
16.
17.
18.
References
Papers of particular interest, published recently, have been
highlighted as:
• Of importance
•• Of major importance
19.
20.
21.
1.
2.
Matsuda M, Shimomura I. Increased oxidative stress in obesity:
implications for metabolic syndrome, diabetes, hypertension, dyslipidemia, atherosclerosis, and cancer. Obesity Research &
Clinical Practice. 2013;7(5):e330–41.
Sies H. Oxidative stress: introduction. oxidative stress: oxidants
and antioxidants. London: Academic; 1991.
22.
Jones DP. Redefining oxidative stress. Antioxid Redox Signal.
2006;8(9–10):1865–79. doi:10.1089/ars.2006.8.1865.
Lushchak VI. Free radicals, reactive oxygen species, oxidative
stress and its classification. Chem Biol Interact. 2014;224:164–
75. doi:10.1016/j.cbi.2014.10.016.
St-Pierre J, Buckingham JA, Roebuck SJ, Brand MD. Topology of
superoxide production from different sites in the mitochondrial
electron transport chain. J Biol Chem. 2002;277(47):44784–90.
doi:10.1074/jbc.M207217200.
Orrenius S, Gogvadze V, Zhivotovsky B. Mitochondrial oxidative
stress: implications for cell death. Annu Rev Pharmacol Toxicol.
2007;47:143–83. doi:10.1146/annurev.pharmtox.47.120505.105122.
Circu ML, Aw TY. Reactive oxygen species, cellular redox systems, and apoptosis. Free Radic Biol Med. 2010;48(6):749–62.
doi:10.1016/j.freeradbiomed.2009.12.022.
Bedard K, Krause KH. The NOX family of ROS-generating
NADPH oxidases: physiology and pathophysiology. Physiol
Rev. 2007;87(1):245–313. doi:10.1152/physrev.00044.2005.
Pacher P, Beckman JS, Liaudet L. Nitric oxide and peroxynitrite in
health and disease. Physiol Rev. 2007;87(1):315–424. doi:10.
1152/physrev.00029.2006.
Kelley EE, Khoo NK, Hundley NJ, Malik UZ, Freeman BA,
Tarpey MM. Hydrogen peroxide is the major oxidant product of
xanthine oxidase. Free Radic Biol Med. 2010;48(4):493–8. doi:
10.1016/j.freeradbiomed.2009.11.012.
Tu BP, Weissman JS. Oxidative protein folding in eukaryotes:
mechanisms and consequences. J Cell Biol. 2004;164(3):341–6.
doi:10.1083/jcb.200311055.
Salvado L, Palomer X, Barroso E, Vazquez-Carrera M. Targeting
endoplasmic reticulum stress in insulin resistance. Trends in
Endocrinology and Metabolism: TEM. 2015;26(8):438–48. doi:
10.1016/j.tem.2015.05.007.
Brown DI, Griendling KK. Regulation of signal transduction by
reactive oxygen species in the cardiovascular system. Circ Res.
2015;116(3):531–49. doi:10.1161/CIRCRESAHA.116.303584.
Fukai T, Ushio-Fukai M. Superoxide dismutases: role in redox
signaling, vascular function, and diseases. Antioxid Redox
Signal. 2011;15(6):1583–606. doi:10.1089/ars.2011.3999.
Margis R, Dunand C, Teixeira FK, Margis-Pinheiro M. Glutathione
peroxidase family—an evolutionary overview. FEBS J.
2008;275(15):3959–70. doi:10.1111/j.1742-4658.2008.06542.x.
Dikalov SI, Harrison DG. Methods for detection of mitochondrial
and cellular reactive oxygen species. Antioxid Redox Signal.
2014;20(2):372–82. doi:10.1089/ars.2012.4886.
Stephens JW, Khanolkar MP, Bain SC. The biological relevance
and measurement of plasma markers of oxidative stress in diabetes
and cardiovascular disease. Atherosclerosis. 2009;202(2):321–9.
doi:10.1016/j.atherosclerosis.2008.06.006.
Sies H. Hydrogen peroxide as a central redox signaling molecule
in physiological oxidative stress: oxidative eustress. Redox Biol.
2017;11:613–9.
Dikalov S, Griendling KK, Harrison DG. Measurement of reactive
oxygen species in cardiovascular studies. Hypertension.
2007;49(4):717–27. doi:10.1161/01.HYP.0000258594.87211.6b.
Vasquez-Vivar J, Hogg N, Pritchard Jr KA, Martasek P,
Kalyanaraman B. Superoxide anion formation from lucigenin:
an electron spin resonance spin-trapping study. FEBS Lett.
1997;403(2):127–30.
Starkov AA. Measurement of mitochondrial ROS production.
Methods Mol Biol. 2010;648:245–55. doi:10.1007/978-160761-756-3_16.
Zielonka J, Zielonka M, Sikora A, Adamus J, Joseph J, Hardy M,
et al. Global profiling of reactive oxygen and nitrogen species in
biological systems: high-throughput real-time analyses. J Biol
Chem. 2012;287(5):2984–95. doi:10.1074/jbc.M111.309062.
Curr Diab Rep (2017) 17: 41
23.
24.
25.
26.
27.
28.
29.
30.
31.
32.
33.
34.
35.
36.
37.
38.
39.
40.
41.
Buege JA, Aust SD. Microsomal lipid peroxidation. Methods
Enzymol. 1978;52:302–10.
Moselhy HF, Reid RG, Yousef S, Boyle SP. A specific, accurate,
and sensitive measure of total plasma malondialdehyde by HPLC.
J Lipid Res. 2013;54(3):852–8. doi:10.1194/jlr.D032698.
Janicka M, Kot-Wasik A, Kot J, Namiesnik J. Isoprostanesbiomarkers of lipid peroxidation: their utility in evaluating oxidative stress and analysis. Int J Mol Sci. 2010;11(11):4631–59. doi:
10.3390/ijms11114631.
Roberts 2nd LJ, Morrow JD. The generation and actions of
isoprostanes. Biochim Biophys Acta. 1997;1345(2):121–35.
Mayne ST. Antioxidant nutrients and chronic disease: use of biomarkers of exposure and oxidative stress status in epidemiologic
research. J Nutr. 2003;133(Suppl 3):933S–40S.
Halliwell B. Why and how should we measure oxidative DNA
damage in nutritional studies? How far have we come? Am J Clin
Nutr. 2000;72(5):1082–7.
Dalle-Donne I, Rossi R, Giustarini D, Milzani A, Colombo R.
Protein carbonyl groups as biomarkers of oxidative stress.
Clinica Chimica Acta. 2003;329(1–2):23–38.
Grune T, Reinheckel T, Davies KJ. Degradation of oxidized proteins in K562 human hematopoietic cells by proteasome. J Biol
Chem. 1996;271(26):15504–9.
Woolley JF, Stanicka J, Cotter TG. Recent advances in reactive oxygen species measurement in biological systems. Trends Biochem Sci.
2013;38(11):556–65. doi:10.1016/j.tibs.2013.08.009.
Baynes JW. Role of oxidative stress in development of complications in diabetes. Diabetes. 1991;40(4):405–12.
Kumashiro N, Tamura Y, Uchida T, Ogihara T, Fujitani Y, Hirose T,
et al. Impact of oxidative stress and peroxisome proliferator-activated
receptor gamma coactivator-1alpha in hepatic insulin resistance.
Diabetes. 2008;57(8):2083–91. doi:10.2337/db08-0144.
Houstis N, Rosen ED, Lander ES. Reactive oxygen species have a
causal role in multiple forms of insulin resistance. Nature.
2006;440(7086):944–8. doi:10.1038/nature04634.
Furukawa S, Fujita T, Shimabukuro M, Iwaki M, Yamada Y,
Nakajima Y, et al. Increased oxidative stress in obesity and its
impact on metabolic syndrome. J Clin Invest. 2004;114(12):
1752–61. doi:10.1172/JCI21625.
Warolin J, Coenen KR, Kantor JL, Whitaker LE, Wang L, Acra
SA, et al. The relationship of oxidative stress, adiposity and metabolic risk factors in healthy black and white American youth.
Pediatric Obesity. 2014;9(1):43–52. doi:10.1111/j.2047-6310.
2012.00135.x.
Al-Aubaidy HA, Jelinek HF. Oxidative DNA damage and obesity
in type 2 diabetes mellitus. European Journal of Endocrinology/
European Federation of Endocrine Societies. 2011;164(6):899–
904. doi:10.1530/EJE-11-0053.
Yokota T, Kinugawa S, Yamato M, Hirabayashi K, Suga T,
Takada S, et al. Systemic oxidative stress is associated with lower
aerobic capacity and impaired skeletal muscle energy metabolism
in patients with metabolic syndrome. Diabetes Care. 2013;36(5):
1341–6. doi:10.2337/dc12-1161.
Karaouzene N, Merzouk H, Aribi M, Merzouk SA, Berrouiguet
AY, Tessier C, et al. Effects of the association of aging and obesity
on lipids, lipoproteins and oxidative stress biomarkers: a comparison of older with young men. Nutrition, Metabolism, and
Cardiovascular Diseases: NMCD. 2011;21(10):792–9. doi:10.
1016/j.numecd.2010.02.007.
Silver AE, Beske SD, Christou DD, Donato AJ, Moreau KL, Eskurza
I, et al. Overweight and obese humans demonstrate increased vascular
endothelial NAD(P)H oxidase-p47(phox) expression and evidence of
endothelial oxidative stress. Circulation. 2007;115(5):627–37. doi:10.
1161/CIRCULATIONAHA.106.657486.
Lefort N, Glancy B, Bowen B, Willis WT, Bailowitz Z, De
Filippis EA, et al. Increased reactive oxygen species production
Page 11 of 13 41
and lower abundance of complex I subunits and carnitine
palmitoyltransferase 1B protein despite normal mitochondrial respiration in insulin-resistant human skeletal muscle. Diabetes.
2010;59(10):2444–52. doi:10.2337/db10-0174.
42. Anderson EJ, Lustig ME, Boyle KE, Woodlief TL, Kane DA, Lin
CT, et al. Mitochondrial H2O2 emission and cellular redox state
link excess fat intake to insulin resistance in both rodents and
humans. J Clin Invest. 2009;119(3):573–81.
43. Abdul-Ghani MA, Jani R, Chavez A, Molina-Carrion M, Tripathy D,
Defronzo RA. Mitochondrial reactive oxygen species generation in
obese non-diabetic and type 2 diabetic participants. Diabetologia.
2009;52(4):574–82. doi:10.1007/s00125-009-1264-4.
44. Codoner-Franch P, Navarro-Ruiz A, Fernandez-Ferri M, ArillaCodoner A, Ballester-Asensio E, Valls-Belles V. A matter of fat:
insulin resistance and oxidative stress. Pediatr Diabetes.
2012;13(5):392–9. doi:10.1111/j.1399-5448.2011.00847.x.
45. Park S, Kim M, Paik JK, Jang YJ, Lee SH, Lee JH. Oxidative
stress is associated with C-reactive protein in nondiabetic postmenopausal women, independent of obesity and insulin resistance. Clin Endocrinol. 2013;79(1):65–70. doi:10.1111/j.13652265.2012.04512.x.
46. Abramson JL, Hooper WC, Jones DP, Ashfaq S, Rhodes SD,
Weintraub WS, et al. Association between novel oxidative stress
markers and C-reactive protein among adults without clinical coronary heart disease. Atherosclerosis. 2005;178(1):115–21. doi:10.
1016/j.atherosclerosis.2004.08.007.
47. Park K, Gross M, Lee DH, Holvoet P, Himes JH, Shikany JM,
et al. Oxidative stress and insulin resistance: the coronary artery
risk development in young adults study. Diabetes Care.
2009;32(7):1302–7. doi:10.2337/dc09-0259.
48. Kanauchi M, Nishioka H, Hashimoto T. Oxidative DNA damage
and tubulointerstitial injury in diabetic nephropathy. Nephron.
2002;91(2):327–9. 58412
49. Shin CS, Moon BS, Park KS, Kim SY, Park SJ, Chung MH, et al.
Serum 8-hydroxy-guanine levels are increased in diabetic patients.
Diabetes Care. 2001;24(4):733–7.
50. Kant M, Akis M, Calan M, Arkan T, Bayraktar F, Dizdaroglu M, et al.
Elevated urinary levels of 8-oxo-2′-deoxyguanosine, (5′R)- and (5′S)8,5′-cyclo-2′-deoxyadenosines, and 8-iso-prostaglandin F2alpha as
potential biomarkers of oxidative stress in patients with prediabetes.
DNA repair. 2016; doi:10.1016/j.dnarep.2016.09.004.
51. Song F, Jia W, Yao Y, Hu Y, Lei L, Lin J, et al. Oxidative stress,
antioxidant status and DNA damage in patients with impaired
glucose regulation and newly diagnosed type 2 diabetes. Clin
Sci. 2007;112(12):599–606. doi:10.1042/CS20060323.
52. Dave GS, Kalia K. Hyperglycemia induced oxidative stress in
type-1 and type-2 diabetic patients with and without nephropathy.
Cell Mol Biol. 2007;53(5):68–78.
53. Bravard A, Lefai E, Meugnier E, Pesenti S, Disse E, Vouillarmet J,
et al. FTO is increased in muscle during type 2 diabetes, and its
overexpression in myotubes alters insulin signaling, enhances lipogenesis and ROS production, and induces mitochondrial dysfunction. Diabetes. 2011;60(1):258–68. doi:10.2337/db10-0281.
54. Bruce CR, Carey AL, Hawley JA, Febbraio MA. Intramuscular
heat shock protein 72 and heme oxygenase-1 mRNA are reduced
in patients with type 2 diabetes: evidence that insulin resistance is
associated with a disturbed antioxidant defense mechanism.
Diabetes. 2003;52(9):2338–45.
55. Ohara M, Fukui T, Ouchi M, Watanabe K, Suzuki T, Yamamoto S,
et al. Relationship between daily and day-to-day glycemic variability
and increased oxidative stress in type 2 diabetes. Diabetes Res Clin
Pract. 2016;122:62–70. doi:10.1016/j.diabres.2016.09.025.
56.• Konopka AR, Asante A, Lanza IR, Robinson MM, Johnson ML,
Dalla Man C, et al. Defects in mitochondrial efficiency and H2O2
emissions in obese women are restored to a lean phenotype with
aerobic exercise training. Diabetes. 2015;64(6):2104–15. doi:10.
41 Page 12 of 13
57.
58.
59.
60.
61.
62.
63.
64.
65.
66.
67.
68.
69.••
70.
71.
2337/db14-1701. This intervention study provides evidence
that impaired mitochondrial bioenergetics in obese women is
restored towards that of lean, insulin-sensitive individuals 12
weeks of aerobic exercise.
Sparks LM, Xie H, Koza RA, Mynatt R, Hulver MW, Bray GA,
et al. A high-fat diet coordinately downregulates genes required
for mitochondrial oxidative phosphorylation in skeletal muscle.
Diabetes. 2005;54(7):1926–33.
Gregersen S, Samocha-Bonet D, Heilbronn LK, Campbell LV.
Inflammatory and oxidative stress responses to high-carbohydrate
and high-fat meals in healthy humans. Journal of Nutrition and
Metabolism. 2012;2012:238056. doi:10.1155/2012/238056.
Tumova E, Sun W, Jones PH, Vrablik M, Ballantyne CM, Hoogeveen
RC. The impact of rapid weight loss on oxidative stress markers and
the expression of the metabolic syndrome in obese individuals. J
Obes. 2013;2013:729515. doi:10.1155/2013/729515.
Ramezanipour M, Jalali M, Sadrzade-Yeganeh H, Keshavarz SA,
Eshraghian MR, Bagheri M, et al. The effect of weight reduction
on antioxidant enzymes and their association with dietary intake of
vitamins A, C and E. Arquivos brasileiros de endocrinologia e
metabologia. 2014;58(7):744–9.
Bougoulia M, Triantos A, Koliakos G. Plasma interleukin-6
levels, glutathione peroxidase and isoprostane in obese women
before and after weight loss. Association with cardiovascular risk
factors. Hormones. 2006;5(3):192–9.
Johnson JB, Summer W, Cutler RG, Martin B, Hyun DH, Dixit
VD, et al. Alternate day calorie restriction improves clinical findings and reduces markers of oxidative stress and inflammation in
overweight adults with moderate asthma. Free Radic Biol Med.
2007;42(5):665–74. doi:10.1016/j.freeradbiomed.2006.12.005.
Chance B, Sies H, Boveris A. Hydroperoxide metabolism in
mammalian organs. Physiol Rev. 1979;59(3):527–605.
Staniek K, Nohl H. H(2)O(2) detection from intact mitochondria as a
measure for one-electron reduction of dioxygen requires a noninvasive assay system. Biochim Biophys Acta. 1999;1413(2):70–80.
Patti ME, Butte AJ, Crunkhorn S, Cusi K, Berria R, Kashyap S,
et al. Coordinated reduction of genes of oxidative metabolism in
humans with insulin resistance and diabetes: potential role of
PGC1 and NRF1. Proc Natl Acad Sci U S A. 2003;100(14):
8466–71. doi:10.1073/pnas.1032913100.
Mootha VK, Lindgren CM, Eriksson KF, Subramanian A, Sihag
S, Lehar J, et al. PGC-1alpha-responsive genes involved in oxidative phosphorylation are coordinately downregulated in human
diabetes. Nat Genet. 2003;34(3):267–73. doi:10.1038/ng1180.
Szendroedi J, Schmid AI, Chmelik M, Toth C, Brehm A, Krssak
M, et al. Muscle mitochondrial ATP synthesis and glucose
transport/phosphorylation in type 2 diabetes. PLoS Med.
2007;4(5):e154. doi:10.1371/journal.pmed.0040154.
Seifert EL, Estey C, Xuan JY, Harper ME. Electron transport
chain-dependent and -independent mechanisms of mitochondrial
H2O2 emission during long-chain fatty acid oxidation. J Biol
Chem. 2010;285(8):5748–58. doi:10.1074/jbc.M109.026203.
Szendroedi J, Yoshimura T, Phielix E, Koliaki C, Marcucci M, Zhang
D, et al. Role of diacylglycerol activation of PKCtheta in lipidinduced muscle insulin resistance in humans. Proc Natl Acad Sci U
S A. 2014;111(26):9597–602. doi:10.1073/pnas.1409229111. This
paper addresses mechanisms responsible for muscle insulin
resistance in humans and supports the hypothesis that
activation of protein kinase Cθ by bioactive diacylglycerol and
subsequent impairment of insulin signaling plays a major role in
the pathogenesis of muscle insulin resistance.
Boden G. Endoplasmic reticulum stress: another link between
obesity and insulin resistance/inflammation? Diabetes.
2009;58(3):518–9. doi:10.2337/db08-1746.
Sharma NK, Das SK, Mondal AK, Hackney OG, Chu WS, Kern
PA, et al. Endoplasmic reticulum stress markers are associated
Curr Diab Rep (2017) 17: 41
with obesity in nondiabetic subjects. J Clin Endocrinol Metab.
2008;93(11):4532–41. doi:10.1210/jc.2008-1001.
72. Boden G, Duan X, Homko C, Molina EJ, Song W, Perez O, et al.
Increase in endoplasmic reticulum stress-related proteins and
genes in adipose tissue of obese, insulin-resistant individuals.
Diabetes. 2008;57(9):2438–44. doi:10.2337/db08-0604.
73. Lenin R, Sankaramoorthy A, Mohan V, Balasubramanyam M.
Altered immunometabolism at the interface of increased endoplasmic
reticulum (ER) stress in patients with type 2 diabetes. J Leukoc Biol.
2015;98(4):615–22. doi:10.1189/jlb.3A1214-609R.
74. Papa FR. Endoplasmic reticulum stress, pancreatic beta-cell degeneration, and diabetes. Cold Spring Harbor Perspectives in
Medicine. 2012;2(9):a007666. doi:10.1101/cshperspect.a007666.
75. Gorlach A, Klappa P, Kietzmann T. The endoplasmic reticulum:
folding, calcium homeostasis, signaling, and redox control.
Antioxid Redox Signal. 2006;8(9–10):1391–418. doi:10.1089/
ars.2006.8.1391.
76. Nowotny K, Jung T, Hohn A, Weber D, Grune T. Advanced glycation
end products and oxidative stress in type 2 diabetes mellitus. Biomol
Ther. 2015;5(1):194–222. doi:10.3390/biom5010194.
77. Zhang M, Kho AL, Anilkumar N, Chibber R, Pagano PJ, Shah
AM, et al. Glycated proteins stimulate reactive oxygen species
production in cardiac myocytes: involvement of Nox2
(gp91phox)-containing NADPH oxidase. Circulation.
2006;113(9):1235–43. doi:10.1161/CIRCULATIONAHA.105.
581397.
78. Dillard CJ, Litov RE, Savin WM, Dumelin EE, Tappel AL. Effects
of exercise, vitamin E, and ozone on pulmonary function and lipid
peroxidation. J Appl Physiol Respir Environ Exerc Physiol.
1978;45(6):927–32.
79. Veskoukis AS, Goutianos G, Paschalis V, Margaritelis NV,
Tzioura A, Dipla K, et al. The rat closely mimics oxidative stress
and inflammation in humans after exercise but not after exercise
combined with vitamin C administration. Eur J Appl Physiol.
2016;116(4):791–804. doi:10.1007/s00421-016-3336-8.
80. Kim KS, Paik IY, Woo JH, Kang BY. The effect of training type on
oxidative DNA damage and antioxidant capacity during threedimensional space exercise. Medical Principles and Practice.
2010;19(2):133–41. doi:10.1159/000273075.
81. Dantas FF, Brasileiro-Santos Mdo S, Batista RM, do Nascimento
LS, Castellano LR, Ritti-Dias RM, et al. Effect of strength training
on oxidative stress and the correlation of the same with forearm
vasodilatation and blood pressure of hypertensive elderly women:
a randomized clinical trial. PLoS One. 2016;11(8):e0161178. doi:
10.1371/journal.pone.0161178.
82. Roh H-T, Cho S-Y, So W-Y. Obesity promotes oxidative stress and
exacerbates blood-brain barrier disruption after high-intensity exercise. J Sport Health Sci. 2016:1–6.
83. Vincent HK, Morgan JW, Vincent KR. Obesity exacerbates oxidative stress levels after acute exercise. Med Sci Sports Exerc.
2004;36(5):772–9.
84. Vincent HK, Vincent KR, Bourguignon C, Braith RW. Obesity
and postexercise oxidative stress in older women. Med Sci
Sports Exerc. 2005;37(2):213–9.
85. Haxhi J, Leto G, di Palumbo AS, Sbriccoli P, Guidetti L, Fantini C,
et al. Exercise at lunchtime: effect on glycemic control and oxidative
stress in middle-aged men with type 2 diabetes. Eur J Appl Physiol.
2016;116(3):573–82. doi:10.1007/s00421-015-3317-3.
86. Parker L, Stepto NK, Shaw CS, Serpiello FR, Anderson M, Hare DL,
et al. Acute high-intensity interval exercise-induced redox signaling is
associated with enhanced insulin sensitivity in obese middle-aged
men. Front Physiol. 2016;7:411. doi:10.3389/fphys.2016.00411.
87. Berdichevsky A, Guarente L, Bose A. Acute oxidative stress can
reverse insulin resistance by inactivation of cytoplasmic JNK. J Biol
Chem. 2010;285(28):21581–9. doi:10.1074/jbc.M109.093633.
Curr Diab Rep (2017) 17: 41
Bianchi VE, Ribisl PM. Reactive oxygen species response to exercise
training and weight loss in sedentary overweight and obese female
adults. Journal of Cardiopulmonary Rehabilitation and Prevention.
2015;35(4):263–7. doi:10.1097/HCR.0000000000000114.
89. Gutierrez-Lopez L, Garcia-Sanchez JR, Rincon-Viquez Mde J, LaraPadilla E, Sierra-Vargas MP, Olivares-Corichi IM. Hypocaloric diet
and regular moderate aerobic exercise is an effective strategy to reduce
anthropometric parameters and oxidative stress in obese patients.
Obesity Facts. 2012;5(1):12–22. doi:10.1159/000336526.
90. Duggan C, Tapsoba JD, Wang CY, Campbell KL, Foster-Schubert K,
Gross MD, et al. Dietary weight loss, exercise, and oxidative stress in
postmenopausal women: a randomized controlled trial. Cancer Prev
Res. 2016;9(11):835–43. doi:10.1158/1940-6207.CAPR-16-0163.
91. Farinha JB, Steckling FM, Stefanello ST, Cardoso MS, Nunes LS,
Barcelos RP, et al. Response of oxidative stress and inflammatory
biomarkers to a 12-week aerobic exercise training in women with
metabolic syndrome. Sports Medicine - Open. 2015;1(1):3. doi:
10.1186/s40798-015-0011-2.
92. Medeiros Nda S, de Abreu FG, Colato AS, de Lemos LS, Ramis
TR, Dorneles GP, et al. Effects of concurrent training on oxidative
stress and insulin resistance in obese individuals. Oxidative Med
Cell Longev. 2015;2015:697181. doi:10.1155/2015/697181.
93.• Gram M, Vigelso A, Yokota T, Helge JW, Dela F, Hey-Mogensen
M. Skeletal muscle mitochondrial H2O2 emission increases with
immobilization and decreases after aerobic training in young and
older men. J Physiol. 2015;593(17):4011–27. doi:10.1113/
JP270211. This study provides evidence that increased
mitochondrial reactive oxygen species production precipitates
the detrimental effects of physical inactivity.
94. Pittaluga M, Sgadari A, Dimauro I, Tavazzi B, Parisi P, Caporossi
D. Physical exercise and redox balance in type 2 diabetics: effects
of moderate training on biomarkers of oxidative stress and DNA
damage evaluated through comet assay. Oxidative Med Cell
Longev. 2015;2015:981242. doi:10.1155/2015/981242.
95. Dincer S, Altan M, Terzioglu D, Uslu E, Karsidag K, Batu S, et al.
Effects of a regular exercise program on biochemical parameters
of type 2 diabetes mellitus patients. The Journal of Sports
Medicine and Physical Fitness. 2016;56(11):1384–91.
96.•• Vinetti G, Mozzini C, Desenzani P, Boni E, Bulla L, Lorenzetti I,
et al. Supervised exercise training reduces oxidative stress and
cardiometabolic risk in adults with type 2 diabetes: a randomized
controlled trial. Scientific Reports. 2015;5:9238. doi:10.1038/
srep09238. This intervention study shows that supervised
exercise training was able to improve cardiorespiratory
fitness, cardiometabolic risk, and oxidative stress status in
individuals with type 2 diabetes.
97. de Oliveira VN, Bessa A, Jorge ML, Oliveira RJ, de Mello MT,
De Agostini GG, et al. The effect of different training programs on
antioxidant status, oxidative stress, and metabolic control in type 2
diabetes. Applied Physiology, Nutrition, and Metabolism =
Physiologie appliquee, nutrition et metabolisme. 2012;37(2):
334–44. doi:10.1139/h2012-004.
98. McNeilly AM, McClean C, Murphy M, McEneny J, Trinick T,
Burke G, et al. Exercise training and impaired glucose tolerance in
obese humans. J Sports Sci. 2012;30(8):725–32. doi:10.1080/
02640414.2012.671952.
99. Kasimay O, Ergen N, Bilsel S, Kacar O, Deyneli O, Gogas D,
et al. Diet-supported aerobic exercise reduces blood endothelin-1
and nitric oxide levels in individuals with impaired glucose tolerance. Journal of Clinical Lipidology. 2010;4(5):427–34. doi:10.
1016/j.jacl.2010.08.001.
100. Wycherley TP, Brinkworth GD, Noakes M, Buckley JD, Clifton
PM. Effect of caloric restriction with and without exercise training
on oxidative stress and endothelial function in obese subjects with
Page 13 of 13 41
88.
101.
102.
103.
104.
105.
106.
107.
108.
109.
110.
111.
112.
113.
114.
type 2 diabetes. Diabetes Obes Metab. 2008;10(11):1062–73. doi:
10.1111/j.1463-1326.2008.00863.x.
Brinkmann C, Blossfeld J, Pesch M, Krone B, Wiesiollek K,
Capin D, et al. Lipid-peroxidation and peroxiredoxinoveroxidation in the erythrocytes of non-insulin-dependent type
2 diabetic men during acute exercise. Eur J Appl Physiol.
2012;112(6):2277–87. doi:10.1007/s00421-011-2203-x.
Krause M, Rodrigues-Krause J, O’Hagan C, Medlow P, Davison
G, Susta D, et al. The effects of aerobic exercise training at two
different intensities in obesity and type 2 diabetes: implications for
oxidative stress, low-grade inflammation and nitric oxide production. Eur J Appl Physiol. 2014;114(2):251–60. doi:10.1007/
s00421-013-2769-6.
Karstoft K, Clark MA, Jakobsen I, Muller IA, Pedersen BK,
Solomon TP, et al. The effects of 2 weeks of interval vs continuous
walking training on glycaemic control and whole-body oxidative
stress in individuals with type 2 diabetes: a controlled,
randomised, crossover trial. Diabetologia. 2016; doi:10.1007/
s00125-016-4170-6.
Lawler JM, Rodriguez DA, Hord JM. Mitochondria in the middle:
exercise preconditioning protection of striated muscle. J Physiol.
2016;594(18):5161–83. doi:10.1113/JP270656.
Yun J, Finkel T. Mitohormesis. Cell Metab. 2014;19(5):757–66.
doi:10.1016/j.cmet.2014.01.011.
Sandstrom ME, Zhang SJ, Bruton J, Silva JP, Reid MB, Westerblad
H, et al. Role of reactive oxygen species in contraction-mediated
glucose transport in mouse skeletal muscle. J Physiol. 2006;575(Pt
1):251–62. doi:10.1113/jphysiol.2006.110601.
Zhang SJ, Sandstrom ME, Aydin J, Westerblad H, Wieringa B,
Katz A. Activation of glucose transport and AMP-activated protein kinase during muscle contraction in adenylate kinase-1 knockout mice. Acta Physiol. 2008;192(3):413–20. doi:10.1111/j.17481716.2007.01767.x.
Merry TL, Dywer RM, Bradley EA, Rattigan S, McConell GK.
Local hindlimb antioxidant infusion does not affect muscle glucose uptake during in situ contractions in rat. J Appl Physiol.
2010;108(5):1275–83. doi:10.1152/japplphysiol.01335.2009.
Merry TL, Wadley GD, Stathis CG, Garnham AP, Rattigan S,
Hargreaves M, et al. N-Acetylcysteine infusion does not affect
glucose disposal during prolonged moderate-intensity exercise in
humans. J Physiol. 2010;588(Pt 9):1623–34. doi:10.1113/
jphysiol.2009.184333.
Zhang SJ, Sandstrom ME, Lanner JT, Thorell A, Westerblad H,
Katz A. Activation of aconitase in mouse fast-twitch skeletal muscle during contraction-mediated oxidative stress. American
Journal of Physiology Cell Physiology. 2007;293(3):C1154–9.
doi:10.1152/ajpcell.00110.2007.
Ristow M, Zarse K, Oberbach A, Kloting N, Birringer M,
Kiehntopf M, et al. Antioxidants prevent health-promoting effects
of physical exercise in humans. Proc Natl Acad Sci U S A.
2009;106(21):8665–70. doi:10.1073/pnas.0903485106.
Safdar A, Little JP, Stokl AJ, Hettinga BP, Akhtar M, Tarnopolsky
MA. Exercise increases mitochondrial PGC-1alpha content and
promotes nuclear-mitochondrial cross-talk to coordinate mitochondrial biogenesis. J Biol Chem. 2011;286(12):10605–17. doi:
10.1074/jbc.M110.211466.
Smith BK, Mukai K, Lally JS, Maher AC, Gurd BJ, Heigenhauser
GJ, et al. AMP-activated protein kinase is required for exerciseinduced peroxisome proliferator-activated receptor co-activator 1
translocation to subsarcolemmal mitochondria in skeletal muscle. J
Physiol. 2013;591(6):1551–61. doi:10.1113/jphysiol.2012.245944.
Irrcher I, Ljubicic V, Hood DA. Interactions between ROS and AMP
kinase activity in the regulation of PGC-1alpha transcription in skeletal muscle cells. American Journal of Physiology Cell Physiology.
2009;296(1):C116–23. doi:10.1152/ajpcell.00267.2007.