See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/339472777

Biochemical investigation of an experimentally induced metabolic syndrome

in rats

Article  in  Indian Journal of Animal Research · February 2020

CITATIONS READS

0 36

2 authors, including:

Nashaat Ghalib
University of Mosul
41 PUBLICATIONS   17 CITATIONS   

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Biochemistry View project

Basic biochemical studies of anemia in lab animal View project

All content following this page was uploaded by Nashaat Ghalib on 25 February 2020.

The user has requested enhancement of the downloaded file.

RESEARCH ARTICLE Indian Journal of Animal Research, Volume 54 Issue 2: 168-172 (February 2020)

Biochemical investigation of an experimentally induced

metabolic syndrome in rats
N.G. Mustafa, M.K. Hasan1 10.18805/ijar.B-1028

ABSTRACT
Metabolic syndrome (MS) is a complex condition characterized by insulin resistance, hyperglycemia, dyslipidemia, and obesity. This
project aims to induce MS in rat and then a demonstration of the main biochemical parameters. In male, Sprague-Dawley rats, MS has
been prompted suitably and relatively by fast (six weeks) approach through a high fructose in drinking water (40%). It has been found
that serum urea, creatinine and total bilirubin raise in MS significantly. Moreover, dyslipidemia has arbitrated via some considerable
lipid profile deviations. In addition, BMI, blood glucose, and insulin monitoring evidently ensure achievement of MS. It is concluded
that a well-established rat model of MS could be employed by a 40% fructose in drinking water.

Key words: Dyslipidemia, Fructose, Insulin resistance, Metabolic syndrome.

INTRODUCTION
Department of Physiology, Biochemistry and Pharmacology,
Metabolic syndrome (MS) or insulin resistance syndrome,
College of Veterinary Medicine, University of Mosul, Mosul, Iraq.
proposed by Gerald Reaven, is the maj or worldwide 1
Department of Biology, College of Science, University of Mosul,
metabolic disorder contributed to the challenge of human
Mosul, Iraq.
health, (Reaven, 1988; Farag et al., 2018). Actually, about
Corresponding Author: N.G. Mustafa, Department of Physiology,
20-30% of the global population may suffer from MS
Biochemistry, and Pharmacology, College of Veterinary Medicine,
(Kennedy et al., 2010), as well as many animal species like
University of Mosul, Mosul, Iraq. Email: nashaat_ghalib@yahoo.com
dogs (Tvarijonaviciute et al., 2012), cattle (Pampori et al.,
2012), laboratory animals (Zhou et al., 2014) and horses it How to cite this article: Mustafa, N.G. and Hasan, M.K. (2020).
may be donated to laminitis (Morgan et al., 2015). Obesity, Biochemical investigation of an experimentally induced metabolic
insulin resistance, hyperglycemia, dyslipidemia and syndrome in rats. Indian Journal of Animal Research, 54(2): 168-172.
hypertension are the main features of MS (Zhou et al., 2014). Submitted: 01-09-2018 Accepted: 25-12-2018 Published: 13-02-2019
The pathophysiology of MS was not absolutely
recognized, therefore a number of suppositions were
mature adipocytes secrete more adipokines (a cytokine
submitted. The crucial one suggests that MS was the
delivered from adipose tissues, contributed to insulin
consequence of the genetics, standard of life and diet
resistance), leptin, adiponectin and proinflammatory
interactions (Martinez et al., 2018), but insulin resistance
cytokines [interleukin-6 (IL-6)] and tumor necrosis factor-
remains the leader of this syndrome (Sudarsanam et al.,
alpha (TNF-alpha). IL-6 and TNF-alpha promote lipolysis
2010; McCracken et al., 2018). On the account of the high
and hepatic fatty acid synthesis. They are also expanding
mass of adipose tissue in MS individual that results from
insulin resistance through negatively direct interaction with insulin
negative energy balance, a great amount of lipids,
receptors. Recently, a proteomic study of certain liver
particularly free fatty acids (FFAs) were liberated into the
proteins, like fructose1,6-bisphosphatase, 1-pyruvate
bloodstream leading to the negative triggering response of
dehydrogenase, fatty acid synthase, Acyl-CoA synthetase 1,
tissues (muscles) to insulin through inhibiting of insulin-
has indicated variable changes in the metabolism of carbohydrates
mediated glucose uptake. Consequently, hyperglycemia
and lipid and related pathways (Hsieh et al., 2016).
evokes the beta-cells of the pancreas to more insulin
Actually, MS is not entirely understood and it is in need
secretion which results in hyperinsulinemia (Sears and Perry,
of much more exploration (McCracken et al., 2018), i.e. little
2015). On the other hand, insulin resistance and elevated
attention has been paid to the assessment of the biochemical
FFAs in blood inhibit glycogenesis develop more
changes correlated to experimentally induced MS in the rat
hyperglycemia and hypertriacylglycerolemia due to the
as a model for this syndrome. In fact, this project aims to
principal role of insulin in the depression of lipolysis. Again,
the induction of MS in the rat by using a simplified way and
insulin resistance results in great lipolysis making more FFAs,
demonstration of the related biochemical profiles.
progressing the deleterious cycle (McCracken et al., 2018).
Kennedy et al., (2010) has proposed that leptin-
deficient mouse (Lep ob/ob) is a vital genetic model for MS MATERIALS AND METHODS
aroused by spontaneous mutation. Nowadays, it can be Experimental animals
experimentally induced in mice. W hereas, Bezerra and Three months old 30 male Sprague-Dawley rats weight 180-
Oliveira (2013) have documented that in the case of obesity, 200g were housed in the standard cages (dimension of

168 Indian Journal of Animal Research

 Biochemical investigation of an experimentally induced metabolic syndrome in rats
43×29×15 cm), under suitable conditions of humidity,
temperature (22±2C) and 12-hour light-dark cycle, in the
animal house unit, college of veterinary medicine, University
of Mosul, during March-April 2018. At the end of experiments,
all animals were sacrificed using diethyl ether (99.5%)
(Scharlau, Spain) anesthesia.
After one week of acclimatization, rats were divided
randomly into three groups (10 rats per group) according to
drinking water; control group on tap water, F20% group, and
F40% group. All three groups diet and water ad libitum and
the chow were standard for rats (NRC, 1995). The
experimental procedures were approved ethically by the
college of veterinary medicine, University of Mosul.
Fructose drinking water
Freshly fructose drinking water was prepared every
alternative day depending on weight to volume formula
(W ong et al., 2016) by using D (-)-fructose 98.5-102%
(Scharlau, Spain), 20g of fructose dissolved in 100ml of tap
water to prepare 20% fructose drinking water (F20%) and
40g of fructose dissolved in 100ml of tap water to prepare
40% fructose drinking water (F40%). Bottles of drinking water
were covered using aluminum foil to avoid fermentation.
Physiological parameters
As far as body weight, an electronic weighing scale with an
accuracy of 0.01 g, has been used whereas body length,
and abdominal circumference has been measured by scale
tape at the beginning and end (six weeks later) of the
experiment. Body length measurement was from nose to
the origin of the tail (Poudyal et al., 2010). Abdominal
circumference was determined about the anterior region of
the abdomen. Anyhow, body weight gain calculated from
values of initial and final weight, while body mass index (BMI)
was found as BMI = body weight (g)/ body length 2 (cm 2)
(Novelli et al., 2007; Nutan and Kochar, 2014). All
measurements were done in anesthetized rats.
Biochemical analysis
The rats were starved overnight, then blood was collected
from the retro-ocular vein by a heparinized capillary tube.
The blood was immediately centrifuged at 3000xg for 15
minutes. The achieved serum was kept at -20C until
biochemical analysis. Fasting serum glucose (FSG), total
protein, albumin, globulin, urea, creatinine, alanine
transaminase (ALT), aspartate transaminase (AST) and total
bilirubin and lipid profiles; cholesterol, TG, VLDL, LDL and
HDL were estimated using automated enzymatic method
(Genri-GS200- China). W hile atherogenic index (AI) was
calculated as:
AI = total cholesterol – HDL / HDL
according to Yokozawa et al., (2006).
Serum insulin was measured using RayBio® rat insulin
ELISA kit (3607 Parkway Lane, GA 30092, USA), whereas
HOMA-IR (homeostatic model assessment of insulin
resistance) and HOMA-β index (homeostatic model
Volume 54 Issue 2 (February 2020)
assessment of β-cell function) were calculated according
to Zhou et al., (2014) just as follows:
HOMA-IR = FSG × FSI (fasting serum insulin) / 22.5
HOMA-β index = 20 × FSI / FSG – 3.5
Statistical analysis
By using Statistical Package for Social Sciences (SPSS,
version 22), all experimental data were examined and
characterized as mean values with their standard deviation
(Mean ± SD) and subjected to one-way analysis of variance
(ANOVA), a probability (p) of 0.05 and 0.001 were
considered as the significant (p0.05) and highly significant
(p0.001), respectively (Sweet and Grace-Martin, 2011).
RESULTS AND DISCUSSION
Laboratory animals, mainly mice and rats, were employed
in MS projects, either transgenic or diet- or drinking water-
induced and because the syndrome in the human and most
animals usually develop from dietary source, therefore MS
was induced in mice and rats by high fructose diet (Wong
et al., 2016) or high fructose drinking water (Al-Agele and
Khudiar, 2016). Biochemical parameters fluctuations
associated with MS induced in rats by fructose in drinking
water have not been thoroughly investigated. Therefore, it
is believed that more investigations are required in this area.
In this project, MS was established in adult male normal
Sprague-Dawley rats (without genetic modification) through
40% (but not 20%) fructose in drinking water. MS was
confirmed by observed obesity, hyperglycemia, insulin
resistance and dyslipidemia (Wong et al., 2016).
Physiological parameters
As can be seen in the Table 1, body weight, body weight
gain and BMI were increased significantly (p0.05) and high
significantly (p0.001) in treated groups when compared with
the initial and with the control group. Remarkable high body
weight gain and BMI must have been referred to obesity
which is undoubtedly one of the cardinal dramatic
phenomena of the MS (Hsieh et al., 2016). Fructose in
drinking water directly contributes to hyperglycemia resulting
in positive energy balance which, by the time, encourage
adipocyte hyperplasia and hypertrophy, as an attempt to
storage “excess fuel”, giving rise to overweight.
Biochemical parameters
Total protein, albumin, globulin and ALT have non-significant
changes in treated groups as illustrated by Table 2. W hereas
urea and creatinine were increased (p0.001) in F40% group
at the end of experiments. There is depression (p0.05)
in AST and a total bilirubin level of both treated group as
compared with their initials and with the control group.
Raised blood urea could result from high production or
impaired excretion, that may be due to various conditions
(Rodwell et al., 2015). W hereas Meyer and Hostetter,
(2007) supposed the strong relationship between high levels
of serum urea and creatinine with insulin resistance,
oxidative stress and free radicals production and systemic
169
 Biochemical investigation of an experimentally induced metabolic syndrome in rats

Table 1: Effects of fructose drinking water (F20% and F40%) on physiological parameters.
Control F20% F40%
Initial 6 weeks Initial 6 weeks Initial 6 weeks
Body weight (g) 189 ± 5.3 254 ± 6.7 193 ± 9.2 271 ± 7.8 A 186 ± 8.8 327 ± 15.4 b,d
Body weight gain (g) - 65 - 68 - 144
(%) (34.39) ± 4.3 (35.23) ± 7.4 (78.68) ± 12.2 d
BMI 0.39 ± 0.02 0.40 ± 0.04 0.36 ± 0.05 0.40 ± 0.05 A 0.35 ± 0.07 0.48 ± 0.09 B,D
Length (cm) 22 ± 1.2 25 ± 1.4 23 ± 1.1 26 ± 2.2 A 23 ± 1.8 26 ± 2.4 B
Abdomen circumference (cm) 17.9 ± 1.5 20.2 ± 1.8 17.3 ± 2.0 20.1 ± 2.1 A 17.5 ± 1.9 22.4 ± 2.1 B
Values are mean ± SD, n = 10 rats for each group.
A-D represent a significant difference (p0.05).
a-d represent a highly significant difference (p0.001).
A, a indicate a significant difference of 6 weeks group as compared to initial group within F20%.
B, b indicate a significant difference of 6 weeks group as compared to initial group within F40%.
D, d indicate a significant difference of 6 weeks of F40% as compared to 6 weeks of control.
Absence of letters means non-significant differences.

Table 2: Effects of fructose drinking water (F20% and F40%) on biochemical parameters.
Control F20% F40%
Initial 6 weeks Initial 6 weeks Initial 6 weeks
Total protein (g/dl) 6.4 ± 0.5 6.2 ± 0.4 6.3 ± 0.6 5.9 ± 0.5 6.4 ± 0.6 5.8 ± 0.5
Albumin (g/dl) 3.8 ± 0.3 3.5 ± 0.4 3.4 ± 0.3 3.3 ± 0.3 3.6 ± 0.5 3.4 ± 0.4
Globulin (g/dl) 2.7 ± 0.1 2.8 ± 0.3 2.9 ± 0.2 2.6 ± 0.1 2.8 ± 0.3 2.7 ± 0.2
Urea (mg/dl) 17 ± 2.2 16.8 ± 2.9 16.6± 3 19 ± 3.4 16.0 ± 3.7 33.2 ± 3.5 b,d
Creatinine (mg/dl) 0.59 ± 0.2 0.55 ± 0.1 0.67 ± 0.1 0.60 ± 0.2 0.71 ± 0.2 2.2 ± 0.3 B,d
ALT (IU/L) 39 ± 5 37 ± 6.7 41 ± 3.8 39 ± 4.5 37 ± 5 42 ± 4.4
AST (IU/L) 132 ± 13 115 ± 9 137 ± 11 97 ± 10 A,C 135 ± 11 78 ± 13 B,D
Total bilirubin (mg/dl) 0.06 ± 0.02 0.07 ± 0.01 0.07 ± 0.03 0.04 ± 0.01 A,C 0.07 ± 0.02 0.02 ± 0.01 b,d
Values are mean ± SD, n = 10 rats for each group.
A-D represent a significant difference (p0.05).
a-d represent a highly significant difference (p0.001).
A, a indicate a significant difference of 6 weeks group as compared to initial group within F20%.
B, b indicate a significant difference of 6 weeks group as compared to initial group within F40%.
C, c indicate a significant difference of 6 weeks of F20% as compared to 6 weeks of control.
D, d indicate a significant difference of 6 weeks of F40% as compared to 6 weeks of control.
Absence of letters means non-significant differences.

Table 3: Effects of fructose drinking water (F20% and F40%) on glucose and insulin.
Control F20% F40%
Initial 6 weeks Initial 6 weeks Initial 6 weeks
Glucose (mmol/l) 6.44 ± 0.40 6.65 ± 0.51 6.31 ± 0.39 6.70 ± 0.60 6.52 ± 0.4 13.30 ± 0.74 b,d
Insulin (µIU/l) 17.8 ± 1.1 18.3 ± 0.8 17.5 ± 1.3 18.1 ± 1.1 18.1 ± 0.9 41.91 ± 2.0 b,d
HOMA-IR 5.09 5.40 4.90 5.38 5.24 24.77 b,d
HOMA-β index 121.08 116.19 124.55 113.12 119.86 85.53 B,D
Values are mean ± SD, n = 10 rats for each group.
A-D represent a significant difference (p0.05).
a-d represent a highly significant difference (p0.001).
B, b indicate a significant difference of 6 weeks group as compared to initial group within F40%.
D, d indicate a significant difference of 6 weeks of F40% as compared to 6 weeks of control.
Absence of letters means non-significant differences.

170 Indian Journal of Animal Research

 Biochemical investigation of an experimentally induced metabolic syndrome in rats

Table 4: Effects of fructose drinking water (F20% and F40%) on lipid profiles.
Control F20% F40%
Initial 6 weeks Initial 6 weeks Initial 6 weeks
Cholesterol (mg/dl) 98 ± 2.4 119 ± 3.6 102 ± 3.0 126 ± 4.3 A 105 ± 2.9 211 ± 6.1 b,d
TG (mg/dl) 64 ± 2.2 61 ± 2.0 67 ± 1.9 71 ± 3.3 66 ± 3.1 149 ± 5.8 b,d
VLDL (mg/dl) 14.1 ± 0.9 17.3 ± 1.1 13.4 ± 0.8 17.9 ± 1.9 A 13.9 ± 0.8 29.7 ± 2.3 b,D
LDL (mg/dl) 29.3 ± 2.0 32.9 ± 2.4 30.1 ± 2.6 35 ± 3.1 A 28.8 ± 2.4 61.5 ± 3.2 b,d
HDL (mg/dl) 44.2 ± 3.0 41.6 ± 3.1 43.8 ± 2.9 39.2 ± 2.9 A 44.4 ± 3.0 34.2 ± 2.0 B,D
Atherogenic index 1.21 ± 0.3 1.84 ± 0.2 1.32 ± 0.3 2.21 ± 0.5 A,C 1.36 ± 0.2 5.15 ± 0.8 b,d
Values are mean ± SD, n = 10 rats for each group.
A-D represent a significant difference (p0.05).
a-d represent a highly significant difference (p0.001).
A, a indicate a significant difference of 6 weeks group as compared to initial group within F20%.
B, b indicate a significant difference of 6 weeks group as compared to initial group within F40%.
C, c indicate a significant difference of 6 weeks of F20% as compared to 6 weeks of control.
D, d indicate a significant difference of 6 weeks of F40% as compared to 6 weeks of control.
Absence of letters means non-significant differences

inflammation characterized by elevated level of TNF-alpha,
IL-6. It could be suggested that hyperglycemia may have
deleteriously effected on the renal tissue impairing the
glomerular function in the clearance of urea. On the other
hand, Al-Agele and Kuduair (2016) believed that renal
damage by fructose may be due to enhancing proteinuria
and renal incompetence to excrete fructose.
The serum creatinine level estimation was utilized to
monitoring of renal failure progressing. High creatinine may
be the byproduct of muscular energy metabolism that shifts
to depend on non-glucose sources to attain energy due to
insulin resistance to which a little amount of glucose can be
utilized. Likewise, the raised level of both serum total bilirubin
may indicate liver injury, while serum AST due to various
tissues damage (Rodwell et al., 2015).
Serum glucose, insulin, and HOMA-IR were elevated
(p0.001) in the group of F40%, Table 3. In the same group,
HOMA-β index was depressed (p0.05). Hyperglycemia that
concomitant MS agrees with almost all results of other
papers dealt with MS. Actually, it has been believed that
insulin resistance evident by higher HOMA-IR is the leading
contributor to the pathogenesis of MS (Mehta et al., 2010).
In insulin resistance, the normal concentration of insulin
cannot give rise to an adequate response or predicted effect,
particularly in adipose tissue, muscles, and liver. As a
compensatory mechanism pancreatic, beta cells produce
more insulin that ensured by depressed HOMA-β index in
our findings, to overcome hyperglycemia (Nolan et al., 2015).
W hen insulins combined with their receptors, tyrosine
phosphorylation of substrates lead to the activation of two
parallel cell signaling pathways; phosphoinositol 3-kinase
(PI3K) and mitogen activated protein kinase (MAPK). In the
case of insulin resistance, MAPK works ordinarily, in contrast
to PI3K which lacks their usual path triggering trouble in
related signaling pathways, ultimately reduced endothelial
NO production that contributing to atherosclerosis (Lawan
et al., 2018).
Lipid profiles were presented in Table 4, although, there
were age-related changes, it is clear that the total
cholesterol, TG, VLDL, LDL were elevated significantly and
HDL decreased as compared with their initial groups and
controlled groups. Anyway, disorders related to adipose
tissue resulting in a defect in the metabolism of FFAs that
worse insulin resistance. However, Rodwell et al., (2015)
suggest that large amount of FFAs encourages more
production of lipoproteins and promote the process of
gluconeogenesis which subsequent in dyslipidemia. Again,
insulin resistance may be the creator of dyslipidemia by
several means. It is familiar that in normal conditions, insulin
suppresses lipolysis in adipocytes. Therefore, when insulin
action is unsuitable, lipolysis will grow to produce more FFAs
in the liver and subsequently the excess amount of TG will
form (Yokozawa et al., 2006). Further, FFAs support
formation of apoB is the chief lipoprotein of VLDL. Eventually,
serum VLDL will be elevated. Insulin contributes to apoB
degradation via the PI3K pathway, but when insulin
resistance exists, more VLDL will be produced. Finally, it is
believed that insulin regulates the activity of lipoprotein lipase
that adjusts VLDL metabolism and clearance so insulin
resistance causes dyslipidemia through excessive VLDL
production. However, VLDL uptake to remnant lipoprotein
and LDL (both accused to the development of
atherosclerosis), while TG in VLDL has a preference to
uptake to HDL, TG enriched HDL consider favored substrate
to the hepatic lipase, therefore HDL clear immediately
leading to low serum HDL level (Chiang et al., 2011). Future
work should focus on the detailed molecular pathophysiology
of induced MS.
CONCLUSION
A well-established rat model of MS could be employed simply
by a 40% fructose in drinking water, that is ready for the
further future work, and it seems that insulin resistance is
essential to MS and related biochemical changes. To the

Volume 54 Issue 2 (February 2020) 171

 Biochemical investigation of an experimentally induced metabolic syndrome in rats
knowledge of the authors, certain parameters as urea,
creatinine and AST were examined in MS for the first time.
REFERENCES
Al-Agele, F.L. and Khudiar, K.K. (2016). Effect of acrylamide and
fructose on some parameters related to metabolic syndrome
in adult male rats. Iraqi J. Vet. Med. 40:125-135.
Bezerra, A.M. and de Oliveira, D.M. (2013). Metabolic syndrome:
molecular basis and reasons for interaction with obesity.
Demetra: Food Nutr. Health. 8: 63-76.
Chiang, J.K., Lai, N.S., Chang, J.K., Koo, M. (2011). Predicting insulin
resistance using the triglyceride-to-high-density lipoprotein
cholesterol ratio in Taiwanese adults. Cardiovasc. Diabetol.
10:93.
Farag, H.A.M., Hosseinzadeh Attar, M.J., Muhammad, B.A.,
Esmaillzadeh, A., El Bilbeisi, A.H. (2018). Comparative
effects of vitamin D and vitamin C supplementations
with and without endurance  physical  activity on metabolic
syndrome patients: a randomized controlled trial. Diabetol.
Metabol. Synd. 10: 80.
Hsieh, C.C., Liao, C.C., Liao, Y.C., Hwang, L.S., Wu, L.Y., Hsieh, S.C.
(2016). Proteomic changes associated with metabolic
syndrome in a fructose-fed rat model. J Food Drug Analysis.
24: 754-761.
Kennedy, A.J., Ellacott, A.K., King, V.L., Hasty, A.H. (2010). Mouse
model of the metabolic syndrome. Dis. Model Mech. 3:
156-166.
Lawan, A., Min, K., Zhang, L., Canfran-Duque, A., Jurczak, M.J.,
Nie, Y., Gavin, T.P.,et al. (2018). Skeletal Muscle–Specific
Deletion of MKP-1 Reveals a p38 MAPK/JNK/Akt Signaling
Node that Regulates Obesity-Induced Insulin Resistance.
Diabetes. 67(4): 624-635.
Martinez, A.G., Lopez-Espinoza, A., Lopez-Uriarte, P.J. Beltran-
Miranda, C.P., et al. (2018). A laboratory environment
previously associated with a palatable diet can result in
overfeeding in rats. Indian J Anim. Res. 52(9): 1267-1270.
McCracken, E., Monaghan, M., Sreenivasan, S. (2018). Pathophysiology
of the metabolic syndrome. Clin. Dermatol. 36: 14-20.
Mehta, N.N., McGillicuddy, F.C., Anderson, P.D., Hinkle, C.C., Shah,
R. (2010). Experimental endotoxemia induces adipose
inflammation and insulin resistance in humans. Diabetes.
59: 172–181.
Meyer, T.W. and Hostetter, T.H. (2007). Uremia. N. Engl. J. Med.
357: 1316-1325.
Morgan, R., Keen, J., McGowan, C. (2015). Equine metabolic
syndrome. Vet. Record. 177(7): 173-179.
Nolan, C.J., Ruderman, N.B., Kahn, S.E., Pederson, O., Prentki,
M. (2015). Insulin resistance as a physiological defense
172
View publication stats
against metabolic stress: Implications for the management
of subsets of type 2 diabetes. Diabetes. 64(3): 673-686.
Novelli, E. L., Diniz, Y. S., Galhardi, C. M., Ebaid, G.M., Rodrigues,
H.G., Mani, F., et al. (2007). Anthropometrical parameters
and markers of obesity in rats. Lab. Animals. 41: 111–119.
NRC: National Research Council. (1995). Nutrient Requirements
of Laboratory Animals. 3rd rev. ed. National Academy
Press, Washington DC.
Nutan and Kochar, G.K. (2014). Impact of diet regime at naturopathy
centers on the nutritional status of non-insulin dependent
diabetes mellitus (NIDDM) patients. Asian J Dairy Food
Res. 33 (1): 62 – 66.
Pampori, Z.A., Raies Haq, M., Ganie, A.A., Singh, A. K. (2012).
Physiology, metabolism and nutritional requirements in
early lactation to augment milk production in cattle. A
review. Agri. Reviews. 33 (3): 181-191.
Poudyal, H., Campbell, F., Brown, L. (2010). Olive leaf extract attenuates
cardiac, hepatic and metabolic changes in high carbohydrate-
high fat-fed rats. J Nutr. 140: 946–953.
Reaven, G.M. (1988). Role of insulin resistance in human disease.
Diabetes. 37: 1595-1607.
Rodwell, V.W., Bender, D.A., Botham, K.M., Kennelly, P.J., Weil,
P.A. (2015). Harper’s Illustrated Review of Biochemistry.
(30th Edn). McGraw-Hill education. USA. pp. 832.
Sears, B. and Perry, M. (2015). The role of fatty acids in insulin
resistance. Lipids Health Dis. 14: 121.
Sweet, S.A. and Grace-Martin, K.A. (2011). Data analysis with
SPSS. A first course in applied statistics. (4 th Edn.).
Pearson Pub Ltd. USA. pp. 288.
Sudarsanam, P., Shanmugam, K.R., Mallikarjuna, K., Sathyavelu
Reddy, K. (2010). Impact of exercise training on carbohydrate
metabolic profiles in the kidney tissue of male albino rats.
Indian J Anim. Res. 44 (1): 1-8.
Tvarijonaviciute, A., Ceron, J.J., Holden, S.L., Cuthbertson, D.J.,
Biourge, V., Morris, P.J., German, A.J. (2012). Obesity-
related metabolic dysfunction in dogs: a comparison with
human metabolic syndrome. BMC Vet. Res. 8:147-155.
Yokozawa, T., Cho, E.J., Sasaki, S., Satoh, A., Okamoto, T., Sei,
Y. (2006). The protective role of Chinese prescription
Kangen-karyu extract on diet-induced hypercholesterolemia
in rats. Biol. Pharmaceut. Bulletin. 29: 760-765.
Wong, S.K., Chin, K.Y., Suhaimi F.H., Fairus, A., Ima-Nirwana, S.
(2016). Animal model of metabolic syndrome. Nutr. Metabol.
30: 65.
Zhou, X., Han, D., Xu, R., Li, S., Wu, H., Qu, C., Wang, F., Wang,
X., Zhao, Y. (2014). A model of metabolic syndrome and
related diseases with intestinal endotoxemia in rats fed a
high fat and high sucrose diet. PLOS One. 9: e115148.
Indian Journal of Animal Research