Differentiation of mesenchymal stem cells derived from adipose tissue into insulin- producing cells using flavonoid compounds of walnut green skin in diabetic model rats
Differentiation of mesenchymal stem cells derived from adipose tissue into insulin- producing cells using flavonoid compounds of walnut green skin in diabetic model rats
Sima Mashayekh,1,*Mahnaz Azarnia,2Esmail Fattahi,3Reza Moghadasali,4
1. Department of Biology, Ayatolla Amoli Branch , Islamic Azad University, Amol , Iran 2. Department of Biology, Kharazmi University, Tehran , Iran 3. Department of Biology, Ayatolla Amoli Branch , Islamic Azad University, Amol , Iran 4. Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR , Tehran , Iran
Introduction: For many years, the use of stem cells to treat diabetes has become a global challenge in regenerative medicine; So that studies with a history of nearly 50 years have proven that stem cells can perform repeated divisions and remain undifferentiated, but if there is a specific stimulus in the environment, they are able to differentiate into different types of dedicated cells; Therefore, this research aimed at the differentiation of adipose-derived mesenchymal stem cells (AD-MSCs) into pancreatic beta cells using the flavonoid compounds of walnut green skin in order to produce or increase insulin hormone at the level of clinical study.
Methods: In this experimental study, the differentiation of AD-MSCs into insulin-producing cells was started under the flavonoid extract obtained from the green skin of walnut with doses of 50 and 100 mg/ml for three weeks. To make rats diabetic, streptozotocin with a dose of 60 mg/kg was injected intraperitoneally. In order to check the differentiation of cells, morphology test, Dithizone staining (DTZ) and insulin measurement were used by ELISA method. At the end, blood was taken from the animals and the levels of glucose, creatine, urea and uric acid and the expression level of specific genes of pancreatic cells namely PDX1 and NGN3 were evaluated by RT-PCR method.
Results: The results showed that under the influence of flavonoid compounds, the differentiated cells were transformed from spindle-shaped to round or clustered cells, and the DTZ-specific staining of the cells was positive and insulin secretion was also proved. The level of serum glucose, creatine, urea and uric acid decreased and the expression level of insulin-producing genes PDX1 and NGN3 showed a significant difference in the experimental groups (P≥0.05).
Conclusion: The findings of this study showed that in addition to the flavonoid compounds in walnut green skin, they can effectively differentiate AD-MSCs into insulin-producing cells. It has even been able to increase the amount of insulin hormone production through the differentiation or production of new pancreatic beta cells.