Angiotensin (1-7) Reduces Apoptosis Of Ins-1 Cells Induced By High Glucose Via Down-Regulating JNK/Caspase-3 Signaling Pathway

Objective:In this study,Culture the INS-1 cells in vitro and intervene these cells in different groups to explore whether angiotensin?1-7?can reduce the apoptosis of INS-1 cells through down-regulating the expression of JNK/Caspase-3 signaling pathway.Become a new target for the prevention and treatment of diabetes.Methods:Making 1640 complete medium according to the cell culture manual.,the rat INS-1insulinoma cell line was inoculated into a 25T flask containing 3ml complete medium and cultured in a constant temperature cell incubator,according to the cell growth status,Change the culture medium once every 24 hours or 48 hours,usingthe trypsin of 0.25%EDTA digeste and passage the cells.All groups of cells were co-cultured for 12 hours under the same culture conditions,and then randomly divided into A,B,C,D,E,5 groups and A group without treatment as control.Group B added Ang?1-7?with final concentration of 10^-5mol/L,Group C added high glucose solution with final concentration of 30mmol/L,Group D added Ang?1-7?with final concentration of 10^-5mol/L and with high glucose level of 30mmol/L?first Ang?1-7?was pretreated for30 minutes.Then add high glucose with 30mmol/L?,Group E added with SP600125 of10^-5mol/L,high glucose of 30 mmol/L,and Ang?1-7?of 10^-5mol/L,?pretreatment with sp600125 for 30 min,then Ang?1-7?for 30 min,then add high sugar solution for 30min?,then co-culture for 24 hours,Repeat 3 times under the same conditions.Annexin-V-PI apoptosis kit was used to detect the apoptosis rate and real-time-PCR was used to detect the JNK,Caspase-3 mRNA content of rat pancreatic islet?cells according to the instructions of the kit.Results:In this study,we observed the effect of angiotensin?1-7?on the apoptosis rate and JNK,Caspase-3 mRNA expression of INS-1 cell line cultured with high glucose.The results were as follows:1.Comparison of apoptosis rate of INS-1 cells:compared with the blank control group,the apoptosis rate of high glucose group increased,Ang?1-7?could decrease the apoptosis rate,but SP600125 could partially block the effect of Ang?1-7?.2.Comparison of the expression of JNK,Caspase-3 mRNA in different groups:compared with the blank control group,Ang?1-7?could partially decrease the expression of JNK,Caspase-3 mRNA in the cells induced by high glucose,while SP600125 could block the effect of Ang?1-7?.Conclusion:Angiotensin?1-7?can reduces the apoptosis of INS-1 cells induced by high glucose via lowering JNK/Caspase-3 signaling pathway.