Professional Documents
Culture Documents
www.elsevier.com/locate/lifescie
Abstract
In the current study, the effect of soy protein and genistein, one of the main isoflavones in soybeans, on blood glucose, lipid profile, and
antioxidant enzyme activities in streptozotocin (STZ)-induced diabetic rats was investigated. Male SpragueDawley rats were divided into
nondiabetic control, STZ, STZ-genistein supplemented group (STZ-G; 600 mg/kg diet), and STZ-isolated soy protein supplemented group (STZISP; 200 g/kg diet). Diabetes was induced by a single injection of STZ (50 mg/kg BW) freshly dissolved in 0.1 mol/L citrate buffer (pH 4.5) into
the intraperitonium. Diabetes was confirmed by measuring the fasting blood glucose concentration 48-h post-injection. The rats with blood
glucose level above 350 mg/dL were considered to be diabetic. Genistein and ISP were supplemented in the diet for 3 weeks.
The supplementation of genistein and ISP increased the plasma insulin level but decreased the HbAIC level of the STZ-induced diabetic rats.
The supplementation of genistein and ISP increased the glucokinase level of the STZ-induced diabetic rats. A significant reduction in glucose-6phosphatase was observed in the groups treated with genistein and ISP in comparison with the diabetic control group. Hepatic superoxide
dismutase, catalase, and glutathione peroxidase activities of the STZ-induced diabetic rats were significantly decreased in comparison with the
control rats. Administering genistein and ISP to the STZ-induced diabetic rats significantly increased those enzyme activities. The concentration of
thiobarbituric acid reactive substances in the STZ-induced diabetic rats was significantly elevated, while the genistein and ISP supplement
decreased it to the control concentration. Genistein and ISP supplements seem to be beneficial for correcting the hyperglycemia and preventing
diabetic complications.
2006 Elsevier Inc. All rights reserved.
Keywords: Soy protein; Genistein; Streptozotocin-induced diabetic rats; Antioxidant enzymes
Introduction
Diabetes mellitus is a major endocrine disorder and growing
health problem in most countries (Gavard et al., 1993; Anderson
et al., 2001). Recently, it was suggested that formation of free
radicals is involved in the pathogenesis of diabetes and the
development of diabetic complications because prolonged
exposure to hyperglycemia increases the generation of free
radicals and reduces capacities of the antioxidant defense system
(Sanders et al., 2001). In spite of the presentation of many
hypoglycemic agents, diabetes and its related complications are
still a major medical problem.
1579
STZ-G
STZ-ISP
Caseina
Isolated soy proteinb
Cornstarch
Sucrose
Corn oil
Cellulose
Mineral mixturec
Vitamin mixtured
Choline bitartrate
DL-methione
Genistein
20.0
55.0
10.0
5.0
5.0
3.5
1.0
0.2
0.3
20.0
54.94
10.0
5.0
5.0
3.5
1.0
0.2
0.3
0.06
20.0
55.0
10.0
5.0
5.0
3.5
1.0
0.2
0.3
b
c
d
Component
Tissue preparations
1580
Fig. 2. Plasma insulin level in experimental groups (n = 8). The insulin values
(mean SD) are expressed as ng/mL. The means sharing a common letter are not
significantly different ( p < 0.05).
Fig. 1. Oral glucose tolerance test in experimental groups (n = 8). The blood
glucose levels (mean SD) are expressed as mg/dL. The means sharing a
common letter are not significantly different ( p < 0.05).
Fig. 3. Glacosylated hemoglobin level in experimental groups (n = 8). The glycosylated hemoglobin values (mean SD) are expressed as mg/g of hemoglobin.
The means sharing a common letter are not significantly different ( p < 0.05).
STZ
STZ-G
Control
b
Glucokinase
0.261 0.023 0.079 0.009 0.103 0.012 0.133 0.012
Glucose-60.165 0.011d 0.457 0.014a 0.396 0.022b 0.302 0.013c
2
phosphatase
STZ
STZ-G
STZ-ISP
Serum
Total cholesterol
2.40 0.16b 3.62 0.20a 2.61 0.09b 2.55 0.08b
(mmol/L)
HDL-cholesterol
1.22 0.05a 0.42 0.05c 0.65 0.06b 0.63 0.06b
(mmol/L)
Triglyceride (mmol/L) 1.11 0.09b 2.03 0.16a 1.16 0.09b 1.15 0.13b
Liver
Cholesterol (mmol/g) 0.13 0.02b 0.23 0.04a 0.16 0.03b 0.15 0.03b
Triglyceride (mmol/g) 0.24 0.02b 0.45 0.05a 0.28 0.03b 0.26 0.03b
Values are mean SD of 8 rats from each group.
Means in the same row not sharing a common superscript are significantly
different ( p < 0.05) between groups.
a,b,c
Table 4
Effect of genistein and isolated soy protein on hepatic SOD, CAT, GSH-Px
activities and TBARS concentration in diabetic rats
STZ-ISP
c
1581
SOD
CAT2
GSH-Px3
TBARS4
STZ
a
STZ-G
c
12.79 0.75
18.60 1.13a
3.01 0.46a
15.12 0.14b
STZ-ISP
b
6.50 0.54
11.06 0.91d
1.23 0.46c
29.53 2.12a
11.00 1.12b
14.30 1.19b
2.27 0.77b
17.91 1.23b
10.52 1.06
12.41 1.13c
2.16 0.46b
19.24 1.06b
diabetic rats than in the control rats (Table 3). The supplementation of genistein and ISP suppressed the increase in the total
cholesterol and triglyceride levels in the serum and liver of the
diabetic rats. The genistein and ISP supplementation decreased in
the serum and hepatic triglyceride levels significantly to almost
the control concentration. The HDL-cholesterol concentration
was also significantly lowered by the induction of diabetes;
however it was higher in the genistein and ISP supplemented
groups compared to the other diabetic group.
Antioxidant enzyme activities and TBARS concentration
The activities of SOD, CAT, GSH-Px and hepatic TBARS
concentration are given in Table 4. SOD and GSH-Px activities
in the liver of the STZ-induced diabetic rats were significantly
decreased compared to the control rats. Administering genistein
and ISP to the STZ-induced diabetic rats significantly increased
those enzyme activities. CAT activity of the rats treated with STZ
was significantly decreased, while genistein and ISP supplement
to the STZ-induced diabetic rats appeared to increase its activity.
The effect was more pronounced in the ISP supplemented group
than in the genistein-supplemented group.
The concentration of TBARS in the STZ-induced diabetic
rats was significantly elevated, the genistein and ISP supplement
decreased it significantly to almost the control concentration.
Serum aminotransferase activity and plasma urea level
The activities of serum AST and ALT and plasma urea level
of control and experimental animals are given in Table 5.
Supplement with the genistein and ISP along with STZ showed
Table 5
Effect of genistein and isolated soy protein on serum ALT and AST activities and
plasma urea level in diabetic rats
Control
ALT (unit/mL)
AST (unit/mL)
Urea (mg/dL)
STZ
c
35.55 1.62
70.60 2.13c
32.62 1.52c
STZ-G
a
63.88 1.90
101.66 3.06a
61.60 2.52a
STZ-ISP
b
42.09 1.06
82.19 1.56b
43.14 1.36b
43.19 1.06b
80.34 1.69b
41.06 2.02b
a,b,c
1582
result indicates that genistein and ISP are capable of ameliorating the impaired diabetic kidney function in addition to its
hypoglycemic control.
Increased activities of AST and ALT are used as indices of
liver damage. When the liver is damaged, these enzymes leak out
of liver cells in large quantities, so their concentrations in the
blood are increased (Tawta et al., 2000). Possible explanation for
the differential effects of genistein and ISP on the activities of
AST and ALT in plasma is that genistein and ISP may inhibit the
liver damage induced by STZ.
Genistein and ISP would appear to contribute to alleviating
the adverse effect of diabetes mellitus by enhancing the lipid
metabolism as well as the hepatic antioxidant defense system.
Genistein and ISP supplements may be beneficial for correcting
the hyperglycemia and preventing diabetic complications. The
ISP appears to be more potent than the genistein. Hence further
biochemical and pharmacological studies are being carried out
to elucidate their mechanism of action.
Acknowledgements
The author is grateful for the financial support provided by
the foundation of Kosin University.
References
Abei, H., 1984. Catalase in vitro. Methods in Enzymology 105, 121126.
Adams, M., Golden, D., Anthon, M., Register, T., Williams, J.K., 2002. The
inhibitory effect of soy protein isolate on atherosclerosis in mice does not
require the presence of LDL receptors or alteration of plasma lipoproteins. 132,
43-49.
Alegre, M., Ciudas, C.J., Fillat, C., Cuinovart, J.J., 1988. Determination of
glucose-6-phosphatase activity using the glucose dehydrogenase-coupled
reaction. Analytical Biochemistry 173, 185189.
Allain, C.C., Poon, L.S., Chen, C.S.G., Richmond, W., Fu, P.C., 1974. Enzymatic
determination of total serum cholesterol. Clinical Chemistry 20, 470475.
Almdal, T.P., Vilstrup, H., 1988. Strict insulin treatment normalizes the organic
nitrogen contents and the capacity of urea-N synthesis in experimental
diabetes in rats. Diabetologica 31, 114118.
Andersen, L., Dinesen, B., Jorgensen, P.N., Poulsen, F., Roder, M.F., 1993.
Enzyme immunoassay for intact human insulin in serum or plasma. Clinical
Chemistry 38, 578582.
Anderson, J.W., Johnstone, B.M., Cook, N., 1995. Meta-analysis of the effects
of soy protein intake on serum ipids. New England Journal of Medicine 333,
276282.
Anderson, R.J., Freedland, K.E., Clouse, R.E., Lustman, P.J., 2001. The
prevalence of comorbid depression in adults with diabetes: a meta-analysis.
Diabetes Care 24, 10691078.
Anthony, M.S., 2000. Soy and cardiovascular disease:cholesterol lowering and
beyond. Journal of Nutrition 30, 662S663S.
Anthony, M.S., Clarkson, T.B., Williams, J.K., 1998. Effects of soy isoflavones
on atherosclerosis: potential mechanisms. American Journal of Clinical
Nutrition 68, 1390S1393S.
Berg, J.M., Tymoczko, J.L., Stryer, L., 2002. Glycolysis and gluconeogenesis,
Biochemistry, 5th ed. W.H. Freeman and Co., New York, pp. 425464.
Clarkson, T.B., 2002. Soy, soy phytoestrogens and cardiovascular disease.
Journal of Nutrition 132, 566S569S.
Crouse, J.R., Morgan, T., Terry, J.G., Ellis, J., Vitolins, M., Burke, G., 1999. A
randomized trial comparing the effect of casein with that of soy protein
containing varying amounts of isoflavones on plasma concentrations of
lipids and lipoproteins. Archives of Internal Medicine 159, 20702076.
Davidson, A.L., Arion, W.J., 1987. Factors underlying significant underestimations of glucokinase activity in crude liver extracts: physiological
1583
1584
Reddi, A.S., Bollineni, J.S., 2001. Selenium-deficient diet renal oxidative stress
and injury via TGF-beta I in normal and diabetic rats. Kidney International
59, 13421353.
Reeves, P.G., Nielsen, F.H., Fahey, G.C., 1993. AIN-93 purified diets for
laboratory rodents: final report of the American institute of nutrition Ad Hoc
writing committee on reformulation of the AIN-76 rodent diet. Journal of
Nutrition 123, 19391951.
Reitman, S., Frankel, S., 1957. A colorimetric method for the determination of
serum glutamic oxaloacetic and glutamic pyruvic transaminase. American
Journal of Clinical Pathology 28, 815.
Sanders, R.A., Rauscher, F.M., Watkins, J.B., 2001. Effects of quercetin on
antioxidant defense in streptozotocin-induced diabetic rats. Journal of
Biochemical and Molecular Toxicology 15, 143149.
Saravanan, B.R., Pugalendi, K.V., 2005. Influence of sesame oil on blood
glucose, lipid peroxidation, and antioxidant status in streptozotocin diabetic
rats. Journal of Medicinal Food 8, 377381.
Skim, F., Lazrek, H.B., Kaaya, A., El-Amri, H., Jana, M., 1999. Pharmcological
studies of two antidiabetic plnts: Globulari alypum and Zygophyllum
gaetulum. Therapia 54, 711715.
Sundaram, R.K., Bhaskar, A., Vijayalingam, S., Viswanatthan, M., Moha, R.,
Shanmugasundaram, K.R., 1996. Antioxidant status and lipid peroxidation