Study On The Inhibition Of Apoptosis Of H9C2 Cardiomyocytes Induced By Hypoxia/Reoxygenation Through P38MAPK Signaling Pathway

Objective: To determine the effect and mechanism of Luhong prescription on ischemia and reperfusion(I/R)-induced myocardial injury and apoptosis in rat H9C2 cells.Methods: H9C2 cells were pretreated with different concentrations of Luhong prescription,and myocardial injury was simulated by ischemia/reperfusion(IRI)in vitro by hypoxia/reoxygenation.The morphology of H9C2 cardiomyocytes was observed by inverted microscope.The content of lactate dehydrogenase(LDH)in the supernatant was detected by dinitrophenylhydrazine colorimetry,and malondialdehyde in cells was detected by thiobarbituric acid colorimetry.MDA)content,the content of superoxide dismutase(SOD)in cells was detected by xanthine oxidation method;CCK-8Experimental analysis of the effect of Luhong prescription on the survival rate of H9C2 cardiomyocytes;apoptosis of cardiomyocytes by flow cytometry;detection of p38 MAPK,phosphorylated p38MAPK(p-p38MAPK),activated cysteine-containing days by Western blot Expression of Cleaved Caspase-3 protein.Results: After modeling,the cells showed shrinkage of the cell membrane under an inverted microscope,the organelles became smaller,and the nucleus shrunk.Compared with the control group,the cell survival rate of the model group,Luhong prescription group and SB203580 group decreased(P <0.05),the LDH level in the culture supernatant,and the MDA content in the cells increased(P <0.05).The content of SOD decreased(P <0.05),and the levels of p-p38 MAPK and caspase-3 in cells were higher than those in the control group(P <0.05).Compared with the model group,both Luhong prescription and SB203580 pretreatment increased cell viability(P <0.05),apoptosis rate decreased significantly(P <0.05),LDH level in culture supernatant,MDA in cells.The content of SOD was lower in the cells than in the model group(P <0.05).The levels of p-p38 MAPK and caspase-3 in the cells were lower than those in the model group(P <0.05).Conclusion: Luhong prescription can reduce oxidative stress injury and apoptosis induced by hypoxia/reoxygenation in H9C2 cardiomyocytes,improve cell survival rate and anti-oxidative stress damage,and its mechanism may be through inhibition of p38 MAPK signal.Activation of the pathway suggests that it has potential application value in the prevention and treatment of myocardial ischemia-reperfusion injury.