The Study On MiR-21-5p Via The PI3K/Akt Pathway Regulating AEC ? Apoptosis Of Rats Induced By H₂O₂

Objective: To verify that miR-21-5p can regulate the PI3K/Akt pathway to antagonize AEC? apoptosis,and play a protective role through studying that miR-21-5p regulate the apoptosis of alveolar type II epithelial cells?AEC??induced by hydrogen peroxide?H₂O₂?.Methods: The subculture AEC? were divided into 6 groups?n=7?: group A?control group:Using PBS liquid treatment?,group B?injury group: 0.5mmol/L H₂O₂ processing?,group C?miR-21-5p overexpression group: miR-21-5p lentivirus overexpression vector was transfected into AEC? 24 h,and induced by 0.5mmol/L H₂O₂ established apoptosis model?,group D?miR-21-5p empty group: without miR-21-5p lentivirus vector was transfected into AEC? 24 h,and induced by 0.5mmol/L H₂O₂ established apoptosis model?,group E?miR-21-5p inhibition vector group: miR-21-5p lentivirus inhibited that vector was transfected into AEC? 24 h,and induced by 0.5mmol/L H₂O₂ established apoptosis model?,group F?PI3K/Akt blocker group: miR-21-5p lentivirus overexpression vector was transfected into AEC? 24 h,adding 25 ?mol /L PI3K/Akt blocking agent LY294002,and induced by 0.5mmol/L H₂O₂ established apoptosis model?.CCK8 detected cell proliferation activity of 0h,12 h,24h,48 h and observed cell proliferation activity of group A,B,C,D,E,F;Flow cytometry detected the apoptosis rate of 0h,12 h,24h,48 h in each group;Real-time fluorescence quantitative PCR detected the miR-21-5p expression of AEC? in each group;Western Blot-Western blot tested the expression of p Akt protein in each group.Results: 1.The cell proliferation activity of CCK8: with the extension of time,the cell proliferation activity of group A gradually increased.The cell proliferation activity in group B,C,D,E,F decreased?P < 0.05?,and the cell proliferation activity of group C in the five groups decreased obviously slowly than the other four groups?P < 0.05?.2.Flow cytometry detecting the early apoptosis rate?FCM?: the apoptosis rate increases with the extension of injury time in each group?P < 0.05?;In 12 h,24 h,48 h test,the cell apoptosis rate group B is higher than that in group A?P < 0.05?,the apoptosis rate in group C is obviously lower than that in group D,E and F?P < 0.05?;At the identical time point,the apoptosis rate in group E and F is higher than that in group D?P < 0.05?.3.Real-time fluorescence quantitative PCR detecting the miR-21-5p m RNA expression of AEC? in each group: miR-21-5 P expression level in group B is lower than that in group A?P < 0.05?;in group C,D,E and F,miR-21-5 P expression level in group C,F is highest?P < 0.05?,and that in group E is the lowest?P < 0.05?.4.Western blot testing the expression of p Akt protein in each group: B group,p Akt protein expression level in group B is higher than that in group A?P <0.05?;in group C,D,E and F,p Akt protein expression level group C is the highest?P <0.05?,that in group F is the lowest?P <0.05?,and that in group D is higher than that in group E?P <0.05?.Conclusion:?1?miR-21-5p can play an anti apoptotic effect on AEC? induced by H₂O₂.?2?miR-21-5p can inhibit AEC? apoptosis through regulating the PI3K/Akt pathway,and play a protective role.