Therapeutic potential of genistein in ovariectomy-induced pancreatic injury in diabetic rats: The regulation of MAPK pathway and apoptosis

Document Type: Original Article

Authors

1 Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

2 Neurosciences Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

3 Department of Histology & Embryology, Tabriz University of Medical Sciences, Tabriz, Iran

Abstract

Objective(s): Genistein, as a phytoestrogen found in legumes, has several biological activities in general and anti-diabetic activity particularly. In this study, we investigated the effect of genistein on proteins involved in β-cell proliferation, survival and apoptosis to further reveal its anti-diabetic potential in the ovariectomized diabetic rat.
Materials and Methods: We used three-month-old female Wistar rats that either underwent ovariectomy (OVX) or received a sham surgery (Sham). In a subsequent series of experiments, OVX rats received high-fat diet and low dose STZ to induce diabetes (OVX.D) and genistein treatment (OVX.D.G). Western blot analysis was used for the assessment of phosphorylation of ERK1/2 and AKT and expression of Bcl-2 and caspase-3 in pancreas tissue. Hematoxylin-Eosin (H&E) staining was used for histopathological assessment.
Results: Genistein induced AKT and ERK1/2 phosphorylation protein expression of Bcl-2 in the pancreas. In addition, genistein suppressed protein level of caspase-3. Administration of genistein significantly improved hyperglycemia in ovariectomized diabetic rat, concomitant with improved islet β-cell morphology and mass.
Conclusion: These findings suggest that the beneficial antidiabetic effect of genistein partially mediated by directly modulating pancreatic β-cell function via activation of the AKT, ERK1/2, and Bcl-2, as cell survival and anti-apoptotic factors, and decreasing of proapoptotic caspase-3.

Keywords


1. Carr MC. The emergence of the metabolic syndrome with menopause. JCEM 2003; 88:2404-2411.

2. Brand JS, van der Schouw YT, Onland-Moret NC, Sharp SJ, Ong KK, Khaw K-T, et al. Age at menopause, reproductive life Span, and Type 2 diabetes risk results from the EPIC-InterAct study. Diabetes Care 2013; 36:1012-61019.

3. Kim C, Edelstein SL, Crandall JP, Dabelea D, Kitabchi AE, Hamman RF, et al. Menopause and risk of diabetes in the Diabetes Prevention Program. Menopause 2011; 18:857.

4. Bonds D, Lasser N, Qi L, Brzyski R, Caan B, Heiss G, et al. The effect of conjugated equine oestrogen on diabetes incidence: the Women’s Health Initiative randomised trial. Diabetologia 2006; 49:459-468.

5. Appiah D, Winters SJ, Hornung CA. Bilateral oophorectomy and the risk of incident diabetes in postmenopausal women. Diabetes Care 2014; 37:725-733.

6. Pirimoglu Z, Arslan C, Buyukbayrak E, Kars B, Karsidag Y, Unal O, et al. Glucose tolerance of premenopausal women after menopause due to surgical removal of ovaries. Climacteric 2011; 14:453-457.

7. Gilbert ER, Liu D. Anti-diabetic functions of soy isoflavone genistein: mechanisms underlying its effects on pancreatic β-cell function. Food  Funct 2013; 4:200-212.

8. Jayagopal V, Albertazzi P, Kilpatrick ES, Howarth EM, Jennings PE, Hepburn DA, et al. Beneficial effects of soy phytoestrogen intake in postmenopausal women with type 2 diabetes. Diabetes Care 2002; 25:1709-1714.

9. Lavigne C, Marette A, Jacques H. Cod and soy proteins compared with casein improve glucose tolerance and insulin sensitivity in rats. Am J Physiol Endocrinol Metab 2000; 278:E491-E500.

10. Park SA, Choi MS, Cho SY, Seo JS, Jung UJ, Kim MJ, et al. Genistein and daidzein modulate hepatic glucose and lipid regulating enzyme activities in C57BL/KsJ-db/db mice. Life Sci 2006; 79:1207-1213.

11. Cheng SY, Shaw NS, Tsai KS, Chen CY. The hypoglycemic effects of soy isoflavones on postmenopausal women.J Womens Health (Larchmt) 2004; 13:1080-1086.

12. Habibi P, Alihemmati A, NourAzar A, Yousefi H, Mortazavi S, Ahmadiasl N. Expression of the Mir-133 and Bcl-2 could be affected by swimming training in the heart of ovariectomized rats. Iran J Bacic Med Sci 2016; 19:381-387.

13. Srinivasan K, Ramarao P. Animal models in type 2 diabetes research: an overview. Indian J Med Res 2007; 125:451.

14.Yousefi H, Alihemmati A, Karimi P, Alipour MR, Habibi P, Ahmadiasl N. Effect of genistein on expression of pancreatic SIRT1, inflammatory cytokines and histological changes in ovariectomized diabetic rat. Iran J Bacic Med Sci 2017; 20:423-429.

15. Nikasa M, Karimi P, Rajavand H, Afshari F, Jafarlou M, Soltanali M. High cholesterol diet increases expression of cholesterol 24-hydroxylase and BACE1 in rat hippocampi: implications for the effect of diet cholesterol on memory. IRCMJ 2016.

16. Lee JS. Effects of soy protein and genistein on blood glucose, antioxidant enzyme activities, and lipid profile in streptozotocin-induced diabetic rats. Life Sci 2006; 79:1578-1584.

17. Skim F, Lazrek H, Kaaya A, El Amri H, Jana M. Pharmacological studies of two antidiabetic plants: Globularia alypum and Zygophyllum gaetulum. Therapie 1998; 54:711-715.

18. Dickson LM, Rhodes CJ. Pancreatic β-cell growth and survival in the onset of type 2 diabetes: a role for protein kinase B in the Akt? Am J Physiol Endocrinol Metab 2004; 287:E192-E198.

19. Acconcia F, Kumar R. Signaling regulation of genomic and nongenomic functions of estrogen receptors. Cancer Lett 2006; 238:1-14.

20. Acconcia F, Ascenzi P, Bocedi A, Spisni E, Tomasi V, Trentalance A, et al. Palmitoylation-dependent estrogen receptor α membrane localization: regulation by 17β-estradiol. Mol Biol Cell 2005; 16:231-237.

21. Wang A, Liu D, Luo J, Suh KS, Moore W, Alkhalidy H. Phytochemical genistein promotes pancreatic beta-cell survival and exerts anti-diabetic effect via GPR30-mediated mechanism (1045.44).  FASEB J 2014; 28:1045.1044.

22. Wijesekara N, Krishnamurthy M, Bhattacharjee A, Suhail A, Sweeney G, Wheeler MB. Adiponectin-induced ERK and Akt phosphorylation protects against pancreatic beta cell apoptosis and increases insulin gene expression and secretion. J Biol Chem 2010; 285:33623-33631.

23. Lingohr MK, Dickson LM, McCuaig JF, Hugl SR, Twardzik DR, Rhodes CJ. Activation of IRS-2—Mediated signal transduction by IGF-1, but not TGF-α or EGF, augments pancreatic β-Cell proliferation. Diabetes 2002; 51:966-976.

24. Fu Z, Zhang W, Zhen W, Lum H, Nadler J, Bassaganya-Riera J, et al. Genistein induces pancreatic β-cell proliferation through activation of multiple signaling pathways and prevents insulin-deficient diabetes in mice. Endocrinology 2010; 151:3026-3037.

25. Liadis N, Murakami K, Eweida M, Elford AR, Sheu L, Gaisano HY, et al. Caspase-3-dependent β-cell apoptosis in the initiation of autoimmune diabetes mellitus. Mol Cell Biol 2005; 25:3620-3629.

26. Duxbury MS, Ito H, Benoit E, Zinner MJ, Ashley SW, Whang EE. RNA interference targeting focal adhesion kinase enhances pancreatic adenocarcinoma gemcitabine chemosensitivity. Biochem Biophys Res Commun 2003; 311:786-792.

27. El-Rayes BF, Ali S, Ali IF, Philip PA, Abbruzzese J, Sarkar FH. Potentiation of the effect of erlotinib by genistein in pancreatic cancer: the role of Akt and nuclear factor-κB. Cancer Res 2006; 66:10553-10559.

28. Aras AB, Guven M, Akman T, Alacam H, Kalkan Y, Silan C, et al. Genistein exerts neuroprotective effect on focal cerebral ischemia injury in rats. Inflam 2015; 38:1311-1321.