The Effect Of Angiotensin (1-7) On High Glucose Induced MIN6 Cell Dedifferentiation And Fox01 Intracellular Localization

Objective:Mouse MIN6 cells were cultured in vitro and Ang(1-7)intervention was given to observe the expression of cell surface factor MafA,Ngn3 and the change of FoxO1 expression position in the cells under confocal microscope using immunofluorescence technique,so as to explore whether Ang(1-7)could improve the differentiation of MIN6+cells induced by high glucoseMethods:1.The mouse MIN6 cells were cultured in a 25 ml culture flask with a DMEN medium prepared according to a certain proportion and placed in a 37?,5%C02 concentration cell incubator.the culture medium was replaced once every 24 hours,and the cells were subcultured when they grew to 80%of the bottle wall.according to the cell growth status,the 3-4 generation cells were selected for experimental operation2.The cells were randomly divided into four groups,the A group was normal control without intervention,and the B group was added mmol/L of high sugar solution prepared in advance;C group was added 25 mmol/L high sugar solution 10-5 mol/L of Ang(1-7)solution,(add Ang(1-7 first)and add high sugar solution after 30 minutes);D group mmol/L 25 high sugar solution 10-5 mol/L of Ang(1-7)10-5mol/L solution(A-779,30 minutes before Ang(1-7)and 30 minutes after high sugar solution);Co-cultured 2.the same conditions for 24 hours3.Use immunofluorescence to observe the expression of MafA,Ngn3 in each group of cells under confocal microscopy;and the location of FoxO1 expression in the cellsResults:The cell expression MafA,non-expression in A group were observed under confocal microscope Ngn3,Fox01 located in nucleus.The expression MafA of cells in B group decreased significantly compared with that in A group,and the ratio of expression Ngn3,Fox01 cytoplasm to nucleus increased.The MafA expression increased in C group compared with the B group,but less than A group,the Ngn3 expression decreased,and the ratio of cytoplasm to nucleus decreased,the difference was statistically significant(P<0.05);D group compared with C group,MafA expression decreased,Ngn3 expression increased,the ratio of cytoplasm to nucleus increased,the difference was statistically significant(P<0.05),and there was no statistical difference in the B groupConclusion:Angiotensin(1-7)improves MIN6 cell dedifferentiation under high glucose induction and attenuates the FoxO1 movement from the nucleus to the cytoplasm under high glucose induction.