The Effect Of Angiotensin (1-7) On High Glucose-induced ? Cells Dedifferentiation By Regulating Min6 Cells Phenotypic Factors

Objective:Culture mouse MIN-6 cells in vitro for group intervention to determine the effect of angiotensin(1-7)on the dedifferentiation of pancreatic islet cells under high glucose stress,providing a new idea for the treatment of diabetes.Methods:On a clean bench for 30 minutes under UV disinfection,inhale MIN-6 mouse islet cells to configured in advance complete medium 3 ml(10% fetal bovine serum,1%penicillin solution,89% RPMI 1640 basal medium mixture),put into a 25 T culture flask after pipetting,and finally incubate at 37? and 5% CO2 cell incubator.Change the liquid for 24 hours,observe the cell growth status under the microscope,adherent cells and uniform growth to 70%-80% is to extend the experiment operation to the fifth generation.Distribute uniformly suspended MIN-6 cells in 6-well plates,mark of distinction A,B,C,and D groups,respectively,as the experimental control group,the high-glucose group,the high-glucose+Ang-(1-7)group,and the high-glucose+Ang-(1-7)+Mas receptor blocker A779 group,and co-cultivate for 24 hours,repeat the above operation 3 times.The expression of pdx-1 protein and Oct4 and Nanog proteins of dedifferentiated progenitor cells were detected by cell immunofluorescence assay.Results:1.Cell morphology: observed under a microscope,the MIN-6 cells in the normalcontrol group were tile-shaped and adhered uniformly to growth;in the hyperglycemia group,Suspended cells were seen,and adherent cells were unevenly distributed,and the morphology is different,becoming shrink and rounder;Compared with the high-glucose group,the Ang(1-7)intervention group showed no obvious floating cells,The morphology is between the control group and the high-sugar group;After the intervention of Ang(1-7)receptor blocker A779,a large number of suspended cells were seen in the bottle,and the adherent cells were greatly reduced,and the distribution was uneven,smaller and rounder,and the shapes were different.2.MIN-6 cells after high-glucose intervention,immunofluorescence detection results showed that: the expression of PDX-1 protein on the surface marker genes of mouse mature ? cells was reduced,while the expression of the progenitor cell surface marker genes Oct4 and Nango increased,This indicates that mature MIN-6 cells are transformed into endocrine progenitor cells under high glucose stress.Compared with the high-glucose group,after Ang(1-7)intervention,pdx-1 protein expression increased,while Oct4,Nango protein expression decreased,It showed that Ang(1-7)intervention made the dedifferentiated progenitor cells differentiate into mature ? cells again.After the intervention of Ang(1-7)receptor blocker A779,the surface markers of?cells and progenitor cells were roughly consistent with the high glucose group.Conclusion:The experiment showed that the mice MIN-6 cells under high glucose stress were given angiotensin(1-7)intervention,which weakened the ability of mature MIN-6 cells to differentiate into endocrine progenitor cells,and improved the progress of diabetes.