Effect of Wogonin on H/R Induced Cardiomyocyte Apoptosis by Regulating the AMPK/NLRP3 Signaling Pathway
Objective:To investigate the effect of wogonin(WOG)on hypoxia/reoxygenation(H/R)-induced cardiomyocyte apoptosis by regulating adenosine monophosphate-activated protein kinase(AMPK)/NOD-like receptor protein 3(NLRP3)signaling pathway.Methods:H9C2 cells were divided into Control group(normal culture),H/R group(H/R induction),L(low dose)-WOG group,M(medium dose)-WOG group,H(high dose)-WOG group(40,80,120 µmol/L WOG were added on the basis of H/R induction),WOG+Compound C group(10 µmol/L AMPK inhibitor Compound C was added on the basis of H-WOG group).The effect of WOG on the proliferation of H9C2 cells was detected by methyl thiazolyl tetrazolium(MTT)assay and 5-ethynyl-2'-deoxyuridine(EdU)staining.The effect of WOG on apoptosis of H9C2 cells was detected by flow cytometry.The levels of serum oxidative stress indexes[malondialdehyde(MDA),reactive oxygen species(ROS),superoxide dismutase(SOD)]and inflammatory factors[interleukin(IL)-1β,IL-18]in H9C2 cells were detected by enzyme-linked immunosorbent assay(ELISA).The expression of AMPK,NLRP3,B cell lymphoma-2(Bcl-2)and Bcl-2 associated X protein(Bax)in H9C2 cells was detected by Western blot(WB).Results:The optical density(OD490),EdU positive cell rate,SOD,AMPK and Bcl-2 of H9C2 cells in H/R group were lower than those in Control group,and the apoptosis rate,IL-1β,IL-18,ROS,MDA,NLRP3 and Bax were higher than those in Control group(P＜0.05).Compared with H/R group,OD490,EdU positive cell rate,SOD,AMPK and Bcl-2 expression in L-WOG group,M-WOG group and H-WOG group increased,while apoptosis rate,IL-1β,IL-18,ROS,MDA,NLRP3 and Bax decreased(P＜0.05).The OD490,EdU positive cell rate,SOD,AMPK and Bcl-2 in WOG+Compound C group were lower than those in H-WOG group,and the apoptosis rate,IL-1β,IL-18,ROS,MDA,NLRP3 and Bax expression were higher than those in H-WOG group(P＜0.05).Conclusion:WOG can inhibit H/R-induced cardiomyocyte apoptosis,which may be relate to the AMPK/NLRP3 signaling pathway.
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