Lidocaine inhibits hypoxia/reoxygenation-induced cardiomyocyte damage by down-regulating microRNA-181a
Objective To investigate the effect of lidocaine on cardiomyocyte H9C2 injury induced by hypoxia/reoxygenation(H/R)by regulating microRNA-181a(miR-181a).Methods H9C2 cells were cultured and an H/R model was established as H/R group;normal cultured cells were used as the control(Con)group.H/R-induced H9C2 cells were treated with 1.0,2.5,5.0,10.0 and 20.0 µmol/L lidocaine and were set as 1.0 μmol/L group,2.5 μmol/L group,5.0 µmol/L group,10.0 µmol/L group and 20.0 μmol/L group,respectively.Anti-miR-NC and anti-miR-181a were transfected into H/R-induced H9C2 cells,included in H/R+anti-miR-NC group and H/R+anti-miR-181a group,respectively.The miR-NC and miR-181a were transfected into H/R-induced H9C2 cells,and then treated with 20 μmol/L lidocaine,which were recorded as H/R+miR-NC+20 µmol/L group and H/R+miR-181a+20 μmol/L group,respectively.The cell activity was detected by MTT assay;flow cytometry was used to detect apoptosis;the expression of Caspase-3 protein was detected by Western blot;real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of miR-181a;the content of malondialdehyde(MDA)and the activities of lactate dehydrogenase(LDH)and and superoxide dismutase(SOD)were detected;the levels of in-flammatory factors[tumor necrosis factor-α(TNF-α),Interleukin-1β(IL-1β),Interleukin-6(IL-6)]were detected by enzyme-linked immunosorbent assay(ELISA).Results Compared with the Con group,the cell activity in the H/R group was significantly decreased(P＜0.05).Compared with the H/R group,the cell activity of 5.0 μmol/L group,10.0 µmol/L group and 20.0 µmol/L group was significantly increased(P＜0.05).Therefore,the experiment was conducted with 20.0 μmol/L lidocaine.Compared with the Con group,apoptosis rate and Caspase-3 protein expression in the H/R group were significantly increased(P＜0.05).Compared with the H/R group,the apoptosis rate and Caspase-3 protein expression in the 20.0 μmol/L group were significantly decreased(P＜0.05).Compared with the Con group,MDA content,LDH activity and inflammatory factor levels in the H/R group were significantly increased,while SOD activity was significantly decreased in the H/R group;compared with the H/R group,MDA content,LDH activity and inflammatory factor level in the 20.0 µmol/L group were significantly decreased,and SOD activity was significantly increased(P＜0.05).Compared with the H/R+anti-miR-NC group,the expression of miR-181a,apoptosis rate and Caspase-3 protein in the H/R+anti-miR-181 a group were significantly decreased(P＜0.05).Compared with the H/R+anti-miR-NC group,the MDA content,LDH activity and inflammatory factor levels in the H/R+anti-miR-181 a group were significantly decreased,while SOD activity was significantly increased(P＜0.05).Compared with the H/R+miR-NC+20.0 µmol/L group,the apoptosis rate,the contents of Caspase-3 protein as well as MDA content,the activity of LDH and the level of inflammatory factors in the H/R+miR-181a+20.0 μmol/L group were significantly increased,and the SOD activity was significantly decreased(P＜0.05).Conclusion Lidocaine inhibits H/R-induced cardiomyocyte H9C2 injury by interfering with the ex-pression of miR-181a.
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维普
