Hindawi

Journal of Diabetes Research

Volume 2018, Article ID 7341242, 8 pages
https://doi.org/10.1155/2018/7341242

Research Article
The Effects of Momordica balsamina Methanolic Extract on
Kidney Function in STZ-Induced Diabetic Rats: Effects on Selected
Metabolic Markers

Anelisiwe Siboto, Ntethelelo Sibiya, Andile Khathi , and Phikelelani Ngubane

School of Laboratory Medicine and Medical Sciences, University of KwaZulu-Natal, Durban, South Africa

Correspondence should be addressed to Phikelelani Ngubane; ngubanep1@ukzn.ac.za

Received 30 November 2017; Revised 10 April 2018; Accepted 14 May 2018; Published 13 June 2018

Academic Editor: Patrizio Tatti

Copyright © 2018 Anelisiwe Siboto et al. This is an open access article distributed under the Creative Commons Attribution
License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is
properly cited.

Background. Studies suggest that Momordica balsamina (intshungu) possesses hypoglycaemic effects. The effects of Momordica
balsamina on diabetic complications such as DN, however, have not been established. Accordingly, this study seeks to
investigate the effects of M. balsamina on kidney function in STZ-induced diabetic rats. Methods. Methanolic extracts (ME) of
M. balsamina’s leaves were used in this study. Short-term effects of M. balsamina methanolic extract on kidney function and
MAP were studied in STZ-induced diabetic rats treated twice daily with M. balsamina methanolic extract (250 mg/kg), insulin
(175 μg/kg, s.c.), and metformin (500 mg/kg) for 5 weeks. Results. M. balsamina methanolic extract significantly increased Na+
excretion outputs in STZ-induced diabetic rats by comparison to STZ-diabetic control rats. M. balsamina methanolic extract
significantly increased GFR in STZ-diabetic rats with a concomitant decrease in creatinine concentration and also reduced
kidney-to-body ratio, albumin excretion rate (AER), and albumin creatinine ratio (ACR). M. balsamina methanolic extract
significantly reduced MAP in STZ-diabetic animals by comparison with STZ-diabetic control animals. These results suggest that
M. balsamina methanolic extract not only lowers blood glucose but also has beneficial effects on kidney function and blood
pressure. Conclusion. These findings suggest that M. balsamina may have beneficial effects on some processes that are associated
with renal derangement in STZ-induced diabetic rats.

1. Introduction Several mechanisms have been reported to contribute to

the development and progression of diabetic nephropathy.
Diabetic nephropathy (DN), as a result of sustained hyper- These mechanisms include oxidative stress; the advanced
glycaemia, affects approximately 20–40% of individuals with glycation end product (AGE) receptor of the AGE pathway
diabetes mellitus [1]. Diabetic nephropathy is a leading and the renin-angiotensin aldosterone system have been
common cause of chronic kidney disease (CKD) and kidney implicated to play a role in the development of diabetic
failure which are the major causes of morbidity and mortality nephropathy [8]. These metabolic flux or pathways are
associated with type 1 diabetes mellitus [2, 3]. The sustained mainly activated by sustained hyperglycaemia. Taken
hyperglycaemia is associated with the structural kidney together, the activation of these pathways cause the intrarenal
abnormalities such as the thickening of the glomerular base- haemodynamic’s changes through the glycosylation of
ment membrane, mesangial expansion, tubular hypertrophy, intrarenal proteins that induce hyperfiltration and glomeru-
and extracellular matrix deposition [4, 5]. These structural lar dysfunction [1, 9].
abnormalities mediate the functional kidney abnormalities Various agents such as oral antihyperglycaemic drugs,
such as declining glomerular filtration rate, electrolyte insulin, and antirenin-angiotensin system (RAS) are cur-
imbalance, increased blood pressure, and increased protein rently available for the prevention and management of DN;
excretion [6, 7]. however, the efficacy of these agents is inadequate [10].
2 Journal of Diabetes Research

Insulin has been shown to normalise glomerular filtration 2.3. Animals. Male Sprague-Dawley rats weighing 250–300 g
rate (GFR) and oxidative status which helps delay the onset were used to carry out the study. The animals were bred
and progression of CKD [11]. However, the bolus adminis- and kept in the Biomedical Research Unit of the University
tration of insulin in high doses is associated with sodium of KwaZulu-Natal under normal laboratory conditions
retention which often leads to the development of hyperinsu- (temperature and humidity) in a 12 h day: 12 h night cycle.
linaemic oedema and elevated blood pressure [12]. Other The animals were allowed access to water ad libitum and
antidiabetic agents such as metformin and insulin secreta- 40 g standard rat chow daily (Meadow Feeds, Pietermaritz-
gogues are at times unable to achieve glycaemic targets and burg, South Africa). All animal experiments were reviewed
may thus be unable to prevent the progression to DN. and accepted by the Animal Research Ethics Committee
Therefore, there is a need to further explore other avenues of the University of KwaZulu-Natal (AREC/049/016DM).
as means of alternative treatment, thus achieving a holistic Before the study commenced, the animals were allowed to
therapy for DN. acclimatise for 5 days in the metabolic cages.
Medicinal plants have been used for treatment of DM
and associated complications [13]. Studies in our laboratory 2.3.1. Diabetes Induction. Type 1 diabetes mellitus was
have demonstrated that medicinal plants such as Syzygium induced in male Sprague-Dawley rats by a single intraperito-
aromaticum and Syzygium cordutum confer renoprotection neal injection of 60 mg/kg STZ in freshly prepared 0.1 M cit-
in streptozotocin- (STZ-) induced diabetic rats [14, 15]. rate buffer (pH 6.3). The control group received the vehicle,
Mormodica balsamina (MB), our plant of interest, is citrate buffer, through the same route. Seven days after the
fairly common and widespread in southern Africa and induction of diabetes, rats with a blood glucose concentration
is closely related to M. charantia [16]. M. charantia has greater than 20 mmol·L−1 were considered diabetic.
been shown to improve renal function by normalising
oxidative status [17]. M. balsamina has been shown to 2.4. Experimental Protocol. The short-term effects of
have hypoglycaemic activity [18]. However, the renopro- M. balsamina were studied in separate groups of nondiabetic
tective effects of M. balsamina have not yet been estab- and STZ-induced diabetic male rats (n = 6) housed individu-
lished. We envisage that evaluation of parameters such as ally in Makrolon polycarbonate metabolic cages (Techniplast,
GFR and electrolyte handling will shed some light on the Labotec, South Africa) for 5 weeks. ME (250 mg/kg, p.o.) was
therapeutic effects of M. balsamina regarding kidney func- administered twice daily at 0900 h and 1500 h by means of a
tion. The aim of the study therefore is to investigate the bulbed steel tube. Animals which received DMSO/saline
effects of M. balsamina on kidney function in STZ-induced (3 mL/kg, p.o.) served as controls, while those given metfor-
diabetic rats. min (500 mg/kg, p.o.) and insulin (200 μg/kg, s.c.) served as
positive controls. Blood glucose measurements were made
once every third day over the period of 5 weeks using the tail
2. Materials and Methods prick method. Glucose was determined using the Elite® gluc-
ometer (Elite (Pty) Ltd., Health Care Division, South Africa).
2.1. Drugs and Chemicals. Drugs were sourced from
Urine volume and urinary glucose concentrations, albumin,
standard pharmaceutical suppliers. All chemicals were of
creatinine, urea, Na+, K+, Cl−, body weight, 24-hour water
analytical grade and were purchased from standard
intake, food consumption, and mean arterial pressure
commercial suppliers.
(MAP) were determined once every third day. MAP was mon-
itored using a noninvasive tail cuff method with photoelectric
2.2. Plant Extraction sensors (IITC Model 31 Computerised Blood Pressure
Monitor, Life Sciences, Woodland Hills, California, USA) as
2.2.1. Plant Material. Momordica balsamina was identified previously described [19]. The unit works with IITC hardware
by a botanist in the Department of Botany at the University system to measure blood pressure and heart rate in conscious
of KwaZulu-Natal, Westville. The plant was harvested at rats. The animals were warmed at ±30°C in an enclosed cham-
the University of KwaZulu-Natal in Durban, South Africa. ber (IITC Model 31 Computerised Blood Pressure Monitor,
The leaves were washed three times with water to remove Life Sciences, Wood land Hills, California, USA) for 30
any residual dirt. The leaves were then grinded to a pulp minutes before taking blood pressure readings. All measure-
using an electric grinder. Thereafter, the pulp was subjected ments were conducted at 0900 h. Creatinine and albumin
to either methanolic or aqueous extraction as previously concentration were measured in plasma and urine samples
described [18]. at the Global Clinical and Viral Laboratory, Amanzimtoti,
South Africa. GFR was calculated using a standard formula
2.2.2. Methanolic Extraction (ME). The pulp (1.15 kg) was from measurements of the plasma and urinary concentrations
extracted by cold percolation with methanol (95%, 6.9 L) of creatinine and urine flow rate in the fifth week.
for 24 h. The methanolic extract was recovered from the mix-
ture and methanol was further added to the plant material for 2.5. Plasma and Tissue Collection. At the end of the 5-week
further extraction. This process was repeated three times to experimental period, all animals were anaesthetised via a
maximise the yield (609 g). The three extracts were pooled gas anaesthetic chamber (100 mg/kg) for 3 minutes using
together and the combined extract was concentrated at halothane. Blood samples were collected by cardiac puncture
reduced pressure (22–26 mmHg) at 45–60°C. into single precooled heparinized containers for biochemical
Journal of Diabetes Research 3

analysis. The blood was centrifuged at 10 ×g in a Hermle 3. Results

Laborechnic GmBH centrifuge (Wehingen, Germany) for
plasma collection. The kidneys were removed, weighed, and 3.1. Energy Balance. Nondiabetic controls, diabetic controls,
then snap frozen in liquid nitrogen. Afterwards, the kidneys and diabetic rats treated with MB, metformin, and insulin
and plasma were stored in an Ultra Bio Freezer (Snijers were assessed for body weight change and 24-hour water
Scientific, Tilburg, Netherlands) at −80°C. consumption weekly for 5 weeks (Table 1). Diabetic control
rats exhibited severe weight loss despite the significantly
2.6. Malondialdehyde (MDA) Measurements. MDA measure- higher weekly intake of food and water when compared
ments were carried out as per a well-established protocol in to the normal nondiabetic control rats (DC versus ND,
our laboratory [15]. Briefly, tissues (50 mg) were homoge- p < 0 05). In contrast, nondiabetic control rats progressively
nized in 500 μL of 0.2% phosphoric acid. The homogenate gained weight from the 2nd week of treatment until the
was centrifuged at 400 ×g for 10 min. Subsequently, 400 μL end of the experimental period. MB-treated (250 mg/kg)
of the homogenate was supplemented with 400 mL 2% phos- STZ-diabetic rats, however, stabilised body weights by
phoric acid and then separated into two glass tubes, each comparison to untreated STZ-diabetic controls from the
receiving equal volumes of the solution. Afterwards, 200 μL 3rd week to the 5th week of the experimental period while
of 7% phosphoric acid was added into both glass tubes significantly decreasing food intake (p < 0 05). The effects of
followed by 400 μL of thiobarbituric acid (TBA)/butylated metformin (500 mg/kg) were similar to those of MB. Admin-
hydroxytoluene (BHT) into one sample test glass tube and istration of insulin (175 μg/kg) improved body weight and
400 μL of 3 mM hydrochloric acid (HCl) into the second decreased food and water intake from the 3rd to the 5th
glass tube which stood as a blank. Thereafter, 200 μL of 1 M week of the experimental period by comparison to untreated
HCl was then added to sample and blank test tubes to estab- STZ-diabetic rats.
lish an acidic pH of 1.5. Both solutions were heated at 100°C
for 15 min and allowed to cool to room temperature. 1.5 ML 3.2. Mean Arterial Pressure (MAP). Figure 1 shows the mean
of butanol was then added to the cooled solution; the sample arterial pressure (MAP) of nondiabetic controls; diabetic
was vortexed for 1 min for rigorous mixing and allowed to controls; and diabetic rats treated with MB, metformin, and
settle until two phases could be differentiated. The top buta- insulin measured weekly over the 5-week experimental
nol layer was transferred to Eppendorf tubes and centrifuged period. Diabetic control rats showed increased MAP over
at 13,200 ×g for 6 min. The samples were aliquoted into a the period of 5 weeks by comparison to nondiabetic rats.
96-well microtiter plate in triplicates, and the absorbance However, the administration of MB stabilised MAP to
was read at 532 nm using a BioTek mQuant spectropho- normalcy from the 3rd week to 5th week by comparison
tometer (Biotek, Johannesburg, South Africa). The absor- to STZ-diabetic controls. The administration of insulin
bance from these wavelengths was used to calculate the (175 μg/kg) also increased MAP by comparison to nondia-
concentration of MDA using Beer’s law. betic rats. The effects of metformin were similar to MB.

3.3. Urine Output. Figure 2 shows the 24 h urine output of

absorbance Final nondiabetic controls, diabetic controls, and diabetic rats
Concentration = treated with MB, metformin, and insulin measured over the
absorption coef f icient 156 mmol−1
5-week period. Diabetic controls showed increased urine out-
1 put throughout the study by comparison to nondiabetic rats.
The administration of MB (250 mg/kg) decreased urine out-
2.7. Glutathione Peroxidase (GPx) and Superoxide Dismutase put from the 4th week to the 5th week of the experimental
(SOD) Activity. Kidney GPx and SOD activity was measured period by comparison to STZ-diabetic rats. The effects of
using assay kits following the manufacturer’s instructions insulin and metformin were similar to those of MB.
(Elabscience and Biotechnology, WuHan). 3.4. Electrolyte Handling. Table 2 shows urinary and plasma
2.8. Aldosterone. Plasma aldosterone was measured using biochemical parameters of nondiabetic controls, diabetic
an ELISA kit following the manufacturer’s instructions controls, and diabetic rats treated with M. balsamina, metfor-
(Elabscience and Biotechnology, WuHan). min, and insulin. On the 5th week of the experimental
period, plasma creatinine concentrations were significantly
2.9. Statistical Analysis. Data is expressed as means ± (p < 0 05) elevated in STZ-diabetic rats in comparison with
standard error of means (SEM). Statistical analysis was con- nondiabetic rats. On the other hand, urinary creatinine con-
ducted using GraphPad Prism and InStat software (version centrations were significantly reduced at the same time point.
5.00, GraphPad Software, San Diego, California, USA). Administration of M. balsamina, however, significantly
Energy balance parameters, MAP, and urine output were reduced plasma creatinine concentrations while significantly
analysed using analysis of variance (ANOVA) for repeated increasing urinary creatinine concentrations by comparison
measures, which was followed by the Bonferroni post hoc with STZ-diabetic rats (p < 0 05). The effects of metformin
test. Terminal parameters were analysed using ANOVA and insulin on plasma creatinine and urinary urea were
followed by the Bonferroni post hoc test, which was used to similar to M. balsamina. Plasma urea concentrations were
analyse differences between the controls and the experimen- significantly (p < 0 05) increased in STZ-diabetic rats, while
tal groups. Values of p < 0 05 indicate statistical significance. plasma albumin was notably reduced by comparison with
4 Journal of Diabetes Research

Table 1: Comparison of food, water intake, and body weight (b.wt) change of nondiabetic controls, diabetic controls, and diabetic rats treated
with MB, metformin, and insulin during a 5-week study (n = 6 in each group). Values are expressed as mean ± SEM.

Time (weeks)
Parameter Treatment
1 2 3 4 5
NC 11.9 ± 3.4 12.2 ± 1.6 12.3 ± 1.7 13.0 ± 0.4 12.9 ± 0.9
DC 27.6 ± 1.2 25.4 ± 1.4 16.7 ± 0.4 28.0 ± 1.1 24.8 ± 0.3
Food intake (g/100 g) MB 10.0 ± 2.1∗ 10.2 ± 2.6∗ 16.4 ± 1.1 26.47 ± 2.4∗ 18.9 ± 0.6
INS 19.2 ± 2.2∗ 18.8 ± 1.9∗ 11.2 ± 0.8 15.5 ± 3.0∗ 14.2 ± 0.9∗
MTF 28.2 ± 0.6∗ 22.7 ± 0.5 21.8 ± 0.5 19.8 ± 0.7 26.8 ± 1.7
NC 30 ± 8.2 35 ± 1.4 32.5 ± 8.5 45 ± 9.6 51.2 ± 7.2
DC 60 ± 9.6 61.3 ± 12.9 95.0 ± 3.7 91.3 ± 3.4 96 ± 1.0
Water intake (mL/100 g) MB 72.5 ± 2.5 67.0 ± 4.1 73.8 ± 8.3 78.7 ± 9.6 76 ± 1.0
INS 73.0 ± 4.4 76.3 ± 9.4 76.3 ± 0.1 76.4 ± 4.8 74.2 ± 2.3
MTF 68.8 ± 3.1 41.3 ± 7.5 45.6 ± 2.5 45.2 ± 2.3 43.8 ± 1.1
NC 44.7 ± 9.4 69.7 ± 5.9 76 ± 8.6 90.7 ± 11.1 92.5 ± 9.9
DC −72 ± 8.6 −87.2 ± 20 −77.5 ± 9.8 −77.5 ± 12 −80 ± 7.4
% b.wt change/week MB −13.8 ± 8∗ −11.1 ± 5∗ −19.5 ± 6.2∗ −15.3 ± 6.1∗ −40.5 ± 11
∗ ∗ ∗ ∗
INS 18.5 ± 13.9 3.2 ± 15.7 30.0 ± 16.2 38.3 ± 18.1 28.3 ± 3.2∗
MTF −23 ± 7.3 −6 ± 6.8 −32 ± 2.9 −11 ± 17.3 −37 ± 3.2
NC 5.1 ± 0.3 5.1 ± 0.4 4.9 ± 0.3 5.8 ± 0.5 5.3 ± 0.1
DC 33.1 ± 1.5# 33.1 ± 0# 33.4 ± 0.2# 33.0 ± 0.8# 33.2 ± 0.2#
Glucose (mmol/L) MB 27.2 ± 0.3 29.1 ± 0.3 24.2 ± 1.9∗ 25.6 ± 0.6∗ 26.4 ± 1.0∗
INS 30.7 ± 0.8 23.2 ± 2.6 21.1 ± 2.3∗ 21.9 ± 1.6∗ 17.1 ± 0.9∗
MTF 28.0 ± 2.1 26.0 ± 1.4 25.7 ± 0.5∗ 24.6 ± 0.9∗ 24.8 ± 0.6∗
#
p < 0 05 by comparison with nondiabetic control and ∗ p < 0 05 by comparison with diabetic control.

150 100

140
훼
훼
80
훼

훼
훼
훼
훼
MAP (mmHG)

130
Urine output (mL)

훼

훼 60
120

110 40

100
20

90
0 1 2 3 4 5 0
(Weeks) 0 1 2 3 4 5
NC INS (Weeks)
DC MTF NC INS
MB DC MTF
MB
Figure 1: Mean arterial blood pressure of nondiabetic controls,
diabetic controls, and diabetic rats treated with M. balsamina, Figure 2: Urine output of nondiabetic controls, diabetic controls,
metformin, and insulin over a period of 5 weeks. Values are and diabetic rats treated with M. balsamina, metformin, and
expressed as mean ± SEM. αp < 0 05 by comparison to nondiabetic insulin over a period of 5 weeks. Values are expressed as mean ± S
control; ★p < 0 05 by comparison with diabetic control. EM. αp < 0 05 by comparison to nondiabetic control; ★p < 0 05 by
comparison with diabetic control.

nondiabetic control rats. However, treatment with M. balsa- albumin in comparison with STZ-diabetic control rats. The
mina significantly (p < 0 05) decreased plasma urea concen- effects of metformin and insulin on plasma urea and plasma
tration while significantly (p < 0 05) increasing plasma albumin were similar to M. balsamina. STZ-diabetic control
Journal of Diabetes Research 5

Table 2: Comparison of plasma biochemical parameters and urinary electrolytes of nondiabetic controls, diabetic controls, and diabetic rats
treated with MB, metformin, and insulin during a 5-week study (n = 6 in each group). Values are expressed as mean ± SEM.

Groups
Parameters
NC DC MB INS MTF
+
Urinary Na (mmol/L) 66.8 ± 1.2 42.3 ± 3.5 #
53.7 ± 1.3∗ 38.2 ± 1.2 66.0 ± 2.9∗
Plasma Na+ (mmol/L) 120.3 ± 3.3 134.0 ± 2.0# 121.5 ± 4.2∗ 133.2 ± 1.3 121.7 ± 1.4∗
Urinary K+ (mmol/L) 45.5 ± 1.7 25.5 ± 2.1# 53.7 ± 1.3∗ 38.2 ± 3.3∗ 55.5 ± 4.1∗
∗
+
Plasma K (mmol/L) 9.9 ± 1.3 7.6 ± 0.6 #
7.9 ± 0.3 9.5 ± 0.7 8.4 ± 0.4∗
Urinary urea (mmol/L) 654.3 ± 6.8 117.8 ± 4.8# 315.3 ± 5.8∗ 356 ± 9.9∗ 351 ± 8.0∗
Plasma urea (mmol/L) 4.9 ± 0.3 17.5 ± 1.3# 7.0 ± 0.6∗ 7.6 ± 0.8∗ 6.4 ± 0.9∗
∗
Urinary albumin (g/L) 1.7 ± 0.2 3.5 ± 0.4 #
3.2 ± 0.7 4.2 ± 0.7 4.7 ± 1.1∗
Plasma albumin (g/L) 16.5 ± 1.0 12.3 ± 0.6# 13.2 ± 0,6 14.3 ± 0.7∗ 14.3 ± 1.3∗
∗ ∗
Urinary creatinine (μmol/L) 9.9 ± 0.7 14.9 ± 0.3 #
10.4 ± 0.2 8.1 ± 0.4 10.8 ± 0.2∗
Creatinine (μmol/L) 32.5 ± 1.3 45.8 ± 1.1# 29.8 ± 1.0∗ 34.8 ± 1.9∗ 28.2 ± 0.8∗
Aldosterone (pg/mL) 0.2 ± 0.02 0.4 ± 0.1# 0.3 ± 0.4∗ 0.4 ± 0.2 0.2 ± 0.1∗
#
p < 0 05 by comparison with nondiabetic control; ∗ p < 0 05 by comparison with diabetic control.

rats treated with M. balsamina, metformin, and insulin

Table 3: Effects of M. balsamina on the kidney-to-body weight ratio
measured after 5 weeks of treatment. The kidney-to-body
after a 5-week study (n = 6 in each group). Values are expressed as
mean ± SEM.
weight ratio was significantly increased in STZ-diabetic
rats by comparison to nondiabetic controls (p < 0 05).
Experimental Final body Kidney Kidney : body The administration of MB, insulin, or metformin did not
groups weight (g) weight (g) weight ratio (%) affect the kidney-tobody weight ratio by comparison to
NC 342.5 ± 9.92 1.11 ± 0.081 0.32 ± 0.02 STZ-diabetic controls.
DC 170 ± 7.39# 1.37 ± 0.09# 0.80 ± 0.02#
3.6. Kidney Function. Table 4 shows factors of kidney func-
MB 209 ± 11.33∗ 1.01 ± 0.031∗ 0.53 ± 0.03∗
tion of nondiabetic controls, diabetic controls, and diabetic
INS 278.5 ± 7.5∗ 1.14 ± 0.14∗ 0.41 ± 0.06∗ rats treated with M. balsamina, metformin, and insulin
∗ ∗
MTF 212.5 ± 0.04 0.97 ± 0.40 0.46 ± 0.023∗ assessed after the 5-week experimental period. STZ-diabetic
control animals showed significantly increased AER, ACR,
#
p < 0 05 by comparison with nondiabetic control; ∗ p < 0 05 by comparison
with diabetic control. and GFR in comparison with nondiabetic animals (p < 0 05).
The administration of M. balsamina, however, significantly
decreased AER, ACR, and GFR on the 5th week of experimen-
showed significantly increased plasma Na+ concentration, tal period (p < 0 05). The effects of insulin were similar to
aldosterone and decreased urinary Na+ concentrations at those of MB.
week 5 by comparison with nondiabetic rats (p < 0 05).
Administration of M. balsamina, however, significantly 3.7. Oxidative Stress Markers. Table 5 shows kidney malon-
(p < 0 05) reduced plasma Na+ and aldosterone concen- diadehyde (MDA) concentration and activities of SOD and
trations, while urinary Na+ concentrations significantly GPx in nondiabetic controls; diabetic controls; diabetic rats
(p < 0 05) increased at week 5 by comparison to STZ- treated with MB, metformin, and insulin measured after 5
induced diabetic rats. The effects of metformin were similar weeks of experimental period. Diabetic control rats had
to M. balsamina. Insulin administration increased plasma significantly increased kidney MDA concentration and had
Na+ and aldosterone concentrations while urinary Na+ a decrease activity of SOD and GPx in comparison with non-
was decreased. Urinary K+ concentrations were significantly diabetic control (p < 0 05). STZ-diabetic rats treated with
(p < 0 05) decreased, while plasma K+ concentrations were MB, insulin (175 μg/kg) or metformin (500 mg/kg) showed
significantly (p < 0 05) elevated in STZ-diabetic rats by significantly decreased MDA levels and increased activity of
comparison to nondiabetic rats. Administration of M. balsa- SOD and GPx in kidneys by comparison with STZ-induced
mina significantly (p < 0 05) increased urinary K+ output diabetic control rats (p < 0 05).
and decreased plasma K+ concentrations on the 5th week of
experimental period. The effects of M. balsamina on urinary
K+ output and plasma K+ were comparable with those of 4. Discussion
metformin and insulin. The current study investigated the effects of M. balsamina on
renal function in STZ-induced diabetic rats. The results
3.5. Kidney Weights. Table 3 shows kidney-to-body weight herein indicate that M. balsamina is effective in ameliorating
ratio of nondiabetic controls, diabetic controls, and diabetic renal fluid and electrolyte handling in STZ-induced diabetic
6 Journal of Diabetes Research

Table 4: Comparison albumin excretion rate (AER), albumin creatinine ratio (ACR), and glomerular filtration rate (GFR) of nondiabetic
controls, diabetic controls, and diabetic rats treated with MB, metformin, and insulin rats after a 5-week study (n = 6 in each group).
Values are expressed as mean ± SEM.

Groups
Parameters
NC DC MB INS MTF
∗ ∗
AER (mg/day) 0.024 ± 0.01 0.29 ± 0.03 #
0.17 ± 0.01 0.15 ± 0.01 0.19 ± 0.03∗
ACR 8.10 ± 013 27.03 ± 1.13# 15.72 ± 1.50∗ 13.48 ± 0.84∗ 19.23 ± 0.28∗
∗ ∗
GFR (mL/min/100 g) 33.5 ± 0.5 24.8 ± 1.3 #
33.7 ± 1.5 37.9 ± 1.3 30.9 ± 2.1∗
#
p < 0 05 by comparison with nondiabetic control; ∗ p < 0 05 by comparison with diabetic control.

intrarenal renin aldosterone system as M. balsamina admin-

Table 5: Effects of MB on MDA concentrations and activities of
istration resulted in decreased concentrations of plasma
SOD and GPx in kidney tissues of non-diabetic controls, diabetic
controls, and diabetic rats treated with MB, metformin, and
aldosterone in comparison to untreated STZ-diabetic rats
insulin. Values are expressed as mean ± SEM. [22, 23]. The results are of importance because diabetes is
well known to lead to high blood pressure that eventually
Parameter measured Treatment Kidney results in cardiovascular complications [24].
NC 0.994 ± 0.020 STZ-induced diabetes is associated with increased kidney
DC 4.21 ± 0.12#
weights, albumin excretion rate (AER), and albumin creati-
nine ratio (ACR) due to the accumulation of extracellular
MDA (nmol·g−1 protein) INS 1.1 ± 0.01∗
proteins and kidney damage [25]. Treatment with M. balsa-
MTF 1.19 ± 0.01∗ mina, however, attenuated kidney hypertrophy as evidenced
MB 1.14 ± 0.01∗ by the reduced kidney to body ratio, AER, and ACR. The
extracellular protein-induced kidney hypertrophy has been
NC 13.43 ± 0.76
shown to cause structural changes such as the thickening of
DC 6.01 ± 0.09#
the glomerular basement membrane, thereby impairing kid-
SOD activity (pg/mL) INS 9.89 ± 0.10∗ ney function which includes fluid and electrolyte imbalance
MTF 10.11 ± 0.02∗ in acute renal failure [25]. Furthermore, the impairment of
MB 10.08 ± 0.05∗ kidney function as a result of the thickening of the glomeru-
lar basement membrane is also associated with elevated cre-
NC 2137.31 ± 0.03 atinine concentration with a reduction in GFR as seen in
DC 1703.491 ± 0.03# the STZ-diabetic rats in this study at the end of the experi-
GPx activity (ng/mL) INS 2062.21 ± 0.02∗ mental period. These changes, however, were attenuated in
MTF 2125.75 ± 0.01∗ the M. balsamina-treated animals as M. balsamina increased
urinary Na+ outputs of STZ-diabetic rats and elevated GFR as
MB 21370.37 ± 0.03∗
assessed by creatinine clearance, suggesting an upregulation
#
p < 0 05 by comparison with nondiabetic control; ∗ p < 0 05 by comparison of renal function. GFR is an important marker of kidney
with diabetic control. function, and treatment-related increases in creatinine clear-
ance indicate improvement of kidney function in experimen-
rats. In addition, M. balsamina reduces oxidative stress and tal animals [11]. In addition, increases in AER and ACR are
MAP in STZ-induced diabetic rats. markers of the thickening basement membrane [25].
Diabetes is a metabolic disorder characterised by hyper- The significant increase in the excretion of Na+ by M. bal-
glycaemia. Previous studies, however, have reported that samina may perhaps occur via the inhibition of increased
medicinal plants have hypoglycaemic effects [14, 20]. In proximal reabsorption of Na+ which is often seen in DM
agreement with previous reports, the use of M. balsamina [26]. Kidneys maintain the optimum chemical composition
crude extract in STZ-induced diabetic rats significantly atten- of body fluids by removing Na+, K+, urea, albumin, and cre-
uated blood glucose concentration in this study. Diabetes atinine [27, 28]. The concentration of these electrolytes, how-
mellitus is also associated with polyphagia, polydipsia, and ever, was increased in the plasma of STZ-diabetic animals
reduction in body weight [21]. Treatment with M. balsamina, possibly due to sustained hyperglycaemia [28]. Treatment
however, lowered food intake while improving body weight. with M. balsamina, however, was shown to increase excre-
Sustained hyperglycaemia has been shown to activate the tion of urea and albumin in STZ-diabetic rats possibly via
intrarenal renin aldosterone system which is thought to the amelioration of hyperglycaemia. Studies have shown that
increase Na+ retention at various sites of the kidney such as improved glycaemic control is beneficial in improving kidney
distal tubule and collecting duct of the nephron [22]. Na+ function [28].
retention, as assisted by increased aldosterone levels, has Hyperglycaemia-induced oxidative stress in diabetes is a
been shown to increase MAP in STZ-diabetic rats as also seen major cause for the development and progression of diabetic
in this study [23]. M. balsamina, however, decreased MAP of microvascular complications such as diabetic nephropathy
STZ-induced diabetic rats possibly via the inhibition of [29]. In diabetes mellitus, hyperglycaemia can simply
Journal of Diabetes Research 7

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