Document Type : Research articles
Authors
1 Department of Biology, Ayatolla Amoli Branch, Islamic Azad University, Amol, Iran
2 Department of Biology, Kharazmi University, Tehran, Iran
3 Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran
Abstract
Background: Differentiation of stem cells into pancreatic beta cells is a global challenge in reconstructive medicine for the treatment of diabetes. Studies have shown that compounds derived from walnut green skin can differentiate stem cells into beta cells. Flavonoid compounds appear to be the main cause of this differentiation.
Objectives: This study examined the effect of flavonoid compounds in walnut green skin on the differentiation of pancreatic beta cells.
Methods: The present study conducted the differentiation of adipose tissue-derived mesenchymal stem cells (AD-MSCs) into insulin-producing cells under flavonoid extract at doses of 50 and 100 mg/ml for three weeks. For diabetic rats, streptozotocin was injected intraperitoneally at a dose of 60 mg/kg. To evaluate cell differentiation, this study utilized morphology, dithizone (DTZ) staining, insulin-proinsulin production, insulin beta-receptor by the immunofluorescence method, and insulin measurement by the ELISA method. Serum glucose, cholesterol, and lipids were measured by enzymatic colorimetric, enzyme kit, and autoanalyzer, respectively. The expression of pancreatic cell-specific genes, including Pancreatic and Duodenal Homeobox 1 (Pdx1) and Neurogenin-3 (Ngn3), was also assessed by real-time PCR.
Results: Differentiated cells transformed from spindle-shaped to circular or clustered cells under the influence of flavonoid compounds with their specific DTZ staining being positive. The expression of insulin-proinsulin markers, beta receptors, and insulin secretion was also confirmed. Decreased blood lipids and glucose, as well as increased expression of Pdx1 and Ngn3 insulin-producing genes, were significant in the treatment groups (P<0.05).
Conclusion: The findings showed that flavonoid compounds could effectively induce the differentiation of AD-MSCs into insulin-producing cells.
Keywords
- Zaree AB, Fallahhossini F, Sharifabady R, Nourozzadeh A, Imani H, Ghoshouni H. the effect of citrallus colocynthis extract on preventing/reducing sterptozotocion-induced diabetes in rat. Kowsar Med J. 2007;12(1):13-20.
- Mohamadi J, Mirzaei A, Azizi A, Rouzbehi A, Delaviz H. The effects of hydro alcoholic extract of juglans regia leaf on histological changes of langerhans islet in diabetic rats model. Iranian South Med J. 2012;15(4):293-301.
- Rahavi H, Hashemi M, Soleimani M, Mohammadi J, Tajik N. Assessing in vitro inhibitory effect of adipose-derived mesenchymal stem cell on C57BL16 diabetic mouse splenocytes prolifereation. RJMS. 2015;132(22):95-106.
- Solali S, Kaviani S, Soleimani M, Zonubi Z. Isolation and characterization of mesenchymal stem cells derived from adipose tissue. Koomesh. 2015;16(4):505-11.
- Esmaeili F, Kharazian N, Houshmand F, Mansourzadeh S. Evaluation of differentiation induction of mesenchymal stem cells in to pancreatic beta cells by methanolic extract of medicago sative L. Daneshvar Medicine Basic and Clinical Research Journal. 2015;22(6):51-65.
- Azizi Shafa M, Ghadimi S, Aeshmati A. Influence of ethanol and methanol extracts of walnut leaf and green hull on saccharomyces cerevisiae. bacillus licheniformis and aspergillus niger in date syrup. Iran. J Nutr Sci Food Technol. 2017;11(4):81-8.
- Asgary S, Rahimi P, Mohzoni L, Kabiri N. Hypoglycemic effects of extract Juglans regia L.leaves on alloxan-induced diabetic rats. J Appl Res Med Aromat Plants. 2010;26(1):30-9. doi: 10.22092/ijmapr.2010.6972
- Rahimipanah M, Hamedi M, Mirzapour M. Analysis of some factors affecting the phenolic compounds extracted from green husk of walnut (Juglsns regia L.). J Appl Res Med Aromat Plants. 2011;27(3):419-30.
- Dastoo R, Bakhshi D, Aliakbar A. The assessment of total phenol,flavonoid,resveratvol content and antioxidant capcity of Vitis Vinife L., Pistacia Vera L., Sambucus nigra L. and Ilex Spiningera Loes. J Medicinal Plants. 2017;5(2):37-48.
- Hosseini E, Karimzadeh K. Anti-diabetic effects of hydroal cohlic juglans regia male flower extract on blood diabetogenized adult male rat. J Birjand Univ Med Sci. 2012;19(2):165-72.
- Wilson B, Liotta LA, Petricoiniii E. Dynamic Protein Pathway activation mapping of adipose-derived stem cell
- differentiation implicates norel regulator of adipocyte differentiation. Mol Cell Proteomics. 2013;12(9):2522-35. doi: 10.1074/mcp.M112.025346. [PubMed: 23750025].
- Garcia MM, Fandel TM, Lin G, Shindel AW, Banie L, Lin CS, et al. Treatment of erectile dysfunction in the obese typy2 diabetic ZDF rat with adipose tissue-derived stem cell.
- J Sexual Med. 2010;1(1):89-98. doi: 10.1111/j.1743-6109.2009.01541.x.
- Moradi A, Mohammadi S, Hamidi Alamdari D. Effect of adipose tissue-derived stem cells on the control of the blood glucose level in diabetic rats. Journal Shahid Sadoughi Univestity of Medical Science. 2015;23(8):717-26.
- Safia M, Samar O, Arwa MA Comparative study of the effect of crude and nanoparticles costus sepciosus on testicular damage assoclated to experimentally induced type 2 diabetes. Pharmacophore. 2019;10(6):99-106.
- Kawakami M, Hirayama A, Tsuchiya K. Ohgaware HM, Nakamura M, Umezawa K. Promotion of beta cell differentiation by the alkaloid conophylline in porcine pancreatic endocrine cells. Biomed Pharamacother. 2010;
- (3):226-31. doi: 10.1016/j.biopha.2009.09.025. [PubMed: 20079600].
- Ali Ahmed AB, Rao AS, Rao MV. In vitro callus and in vivo leaf extract of Gymnema sylvestre stimulate β-cells regeneration and anti-diabetic activity in Wistar rats. Phytomedicine. 2010;17(13):1033-9. doi: 10.1016/j.phymed.2010.03.019. [PubMed: 20537514].
- Bnouham M, Ziyyat A, Mekhfi H, Tahri A, Abdelkhaleq L. Medicinal plants with potential antidiabetic activity-A review of ten years of berbal medicine research (1990-2000). Diabetes Metab J. 2006;14(1):1-8. doi: 10.1159/000497588.
- Han MK. Epigallo catechin gallate, a constituent of green tea, suppresses eytokine-induced pancreatic beta-cell damage. Exp Mol Med. 2003;35(2):136-9. doi: 10.1038/emm.2003.19. [PubMed: 12754418].
- Divband KH, Komili GH, Saeidi-Neek F. Effects of wolnut leaves aqueous extract on blood sugar and serum lipids in diabetic rats. J Birjand Univ Med Sci. 2010;17(1):11-18.
- Bazi Shad A, Miri HR, Esmaeilzadeh Bahabadi S, Hajinezhad MR, Dahmardeh Ghale No F, Sabori H, et al .The effect of hydro-alcoholic extract of prosopis farcta on weight,blood glucose and gene expression of pyruvate kinase in diabetic rat (Type I). Journal of Health Chimes. 2017;4(4):1-9.
- Hedayati M, Pouraboli I, Pouraboli B, Dabiri S, Javadi A. Effects of Otostegia persica extract on serum levels of glucose and morphology of pancreas in diabetic rats. Koomesh. 2012;13(2):201-8.
- Miller CJ, Dunn EV, Hashim IB. Glycemic index of 3varieties of dates. Saudi Med J. 2002;23(5):356-538. [PubMed: 12070575].
- Sharma S, Kulkarni SK, Chopra K. Curumin.the active principle of turmeric (curcuma longa).Ameliorates diabetic hephropathy in rats. Clin Exp Pharmaco Physiol. 2006;33(10):940-5. doi: 10.1111/j.1440-1681.2006.04468.x. [PubMed: 17002671].
- Arddeshiri M, Zia Jahromi N. The effect of chartine on expression of Pdx1gene in streptozotocin-induced diabetic rats. RJMS. 2019;26(6):44-53.
- Hussain F, Tahira S. Antidiabetic evalution of momordica charantia L fruit extracts. West Indian Med J. 2014;63(4):299-99. doi: 10.7727/wimj.2013.180.
- Velayuthom R, Sankaradoss N, Ahamed KF. Protective effect of tannins from ficus racemose in hyper cholesterolemia and diabetes induced vascular tissue damage in rats. Asian
- Pac J Trop Med. 2012;5(5):367-73. doi: 10.1016/S1995-7645(12)60061-3. [PubMed: 22546653].
- Xu X, D´Hoker J. Stange G, Stange G, Bonne S, De Leu N, et al. Beta cells can be generated from endogenous progenitors in injured adult mouse pancreas. Cell. 2008;132(2):197-207. doi: 10.1016/j.cell.2007.12.015. [PubMed: 18243096].
- Vetere A, Marsich E, Di Piazza M, Koncan R, Micali F, Paoletti S. Neurogenin3 triggers beta-cell differentiation of retinoin acid-derived endoderm cells. Biochem J. 2003;371(3):831-41. doi: 10.1042/BJ20021524. [PubMed: 12529176].
- Gomez DL, O´Driscoll M, Sheets TP, Hruban RH, Oberholzer J, McGarrigle JJ, et al . Neurogenin3 expressing cells in the human exocrine pancreas have the capacity for endocrine cell fate. Plos One. 2015;10(8):1-26. doi: 10.1371/journal.pone.ol33862. [PubMed: 26288179].
- Rukstalis JM, Habener JF. Neurogenin3: amaster regulator of pancreatic Islet differentiation and regeneration. Islets. 2003;1(3):177-84. doi: 10.4161/isl.1.3.9877. [PubMed: 21099270].
- Limbert C, Path G, Ebert R, Rothhammer V, Kassem M, Jakob F, et al. PDX1 and NGN3 mediated in vitro reprogramming of human bone marrow-derived mesenchemal stromal cells into pancreatic endocrine lineages. Cytotherapy. 2011;13(7):802-13. doi: 10.3109/14653249.2011.571248. [PubMed: 21506889].
- Miyazaki S, Yamato E, Miyazaki J. Regulated expression of pdx-1 promotes in vitro differentiation of insulin-producing cells from embryonic stem cells. Diabetes. 2004;53(4):1030-7. doi: 10.2337/diabetes.53.4.1030. [PubMed: 15047618].
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