Angiotensin (1-7) Improves Islet β Cell Dedifferentiation Through The Wnt/β-Catenin Signaling Pathway

Objective:The relative expression of specific markers PDX1,MafA,Oct4 and Nanog on mouse MIN6 cell surface under different intervention conditions was semi-quantitatively analyzed by immunofluorescence technique.Furthermore,the effects of Ang(1-7)and Wnt/β-catenin signaling pathways on the dedifferentiation of islet β cells and the relationship between them were discussed,providing a new entry point for the prevention and treatment of diabetes.Methods:1.Add mouse MIN6 cells to the 3ml complete medium prepared in advance,blow evenly and place them in the culture flask,incubate in a 37 °C,5% CO2 cell culture incubator,change the liquid for 24 hours,if the cell growth state is good,start the subculture when it is uniformly adhered to about 80%,and select the 3rd-4th generation of cultured excellent cells according to the detailed experimental steps.2.The cells were randomly divided into normal control group,high sugar group,high sugar + Ang(1-7)group,high sugar + CHIR99021 group,high sugar + Ang(1-7)+CHIR99021 group,high sugar + Ang(1-7)+ A779 group,and intervened for 48 hours under unified culture conditions.3.The expression of cell-specific surface markers PDX1,MafA,Oct4 and Nanog in each group was observed by immunofluorescence technique.Results:Immunofluorescence data showed that compared with the high glucose group,the expression of β cell surface specific maturation markers PDX1 and MafA was significantly increased after treatment with Ang(1-7)alone,and the expression of endocrine progenitor cell markers Oct4 and Nanog was significantly decreased;However,the effect of Ang(1-7)receptor antagonist A779 was significantly reduced;After CHIR99021 alone,the expression of PDX1 and MafA decreased significantly,while the expression of Oct4 and Nanog increased significantly;However,the effect of Wnt/β-catenin was partially eliminated by CHIR99021 and Ang(1-7),and the expression of these factors was reversed to improve the dedifferentiation of islet β cells.Conclusion:1.High glucose can activate the Wnt/β-catenin signaling pathway to promote the dedifferentiation of β cells induced by high glucose;2.Ang(1-7)inhibits the activity of Wnt/β-catenin signaling pathway and effectively weakens the dedifferentiation of islet β cells.