| PurposeThe aim of this study was to investigate the expression status of apoptosisrelated proteins Bcl-2,Bax and Caspase-3 in a model of myocardial ischemia-re perfusion injury and the effect of sakubatril/valsartan intervention.The model was constructed using the left anterior descending branch ligation method to further investigate the relationship between myocardial ischemia-reperfusion injury and apoptosis,as well as the mechanism of the effect of sakubatril/valsartan on myocardial ischemia-reperfusion injury.MethodNinety male rats were randomly divided into three groups: Sham-operatio group(Sham group)(n=30),myocardial ischemia-reperfusion group(I/R group)(n=30),myocardial ischemia-reperfusion + shakobactril/valsartan group(I/R+Entresto group)(n=30).The three groups were sacrificed immediately,4 weeks and 8 wee ks later,respectively.The changes of myocardial structure and morphology were observed by HE staining.The expression levels of Bcl-2,Bax and Caspase-3 in each group at different time were detected by real-time quantitative PCR and Western blotting.Through the use of statistical methods to study whether the changes in these indicators have statistical significance.Result1.Histomorphological changes in the myocardium of rats in each group In the myocardial cells taken from the Shame group,there was no abnormal disorder in the arrangement of structure,the fibrous structure was intact,the transverse lines were clear,the myocardial cytoplasm was evenly stained,no cell swelling was found,and the inflammatory cell infiltration was not obvious.Compared to the Shame group,the myocardial tissue from the I/R group was significantly more damaged at weeks 4 and 8 than at the immediate time of sampling,with blurred myocardial cell structure,poorly defined borders,marked intracellular oedema,uneven nuclear staining and disorganised myocardial fibre alignment,which was significantly better at week 8 than at week 4.In the I/R+Entresto group of rats,swelling of the cardiomyocytes was observed at week 4,with a still intact cell outline,a small set of nuclei at the edge of the cells and a lightly darkened cytoplasm with indistinct margins.The changes in myocardial tissue da mage were more severe than in the immediate sampling group,while myocardial tissue damage had largely recovered by week 8.2.Bcl-2,Bax and Caspase-3 protein and mRNA expression levels(1)Intra-group comparison:There were no significant differences in the m RNA and protein expressions of Bcl-2,Bax and Caspase-3 in Sham group at immediate,4th and 8th week(P>0.05).At 4 weeks,the expression levels of Bax and Caspase-3 in I/R group were the highest,and the difference was statistically significant compared with the immediate group and the rats at 8 weeks(P<0.05).That is,at the 4th week,the expression levels of Bax and Caspase-3 m RNA and protein in I/R group were the highest,while at the 8th week,the expressions of Bax and Caspase-3 m RNA and protein in I/R group were lower than those immediately collected.And the difference was significant(P<0.05).At 4 weeks,the expression level of Bcl-2 in I/R group was the lowest,which was lower than that in immediate group and 8 weeks group,and the difference was statistically significant(P<0.05).That is,the expression level of Bcl-2 m RNA and protein was the lowest at the 4th week,and the expression level of Bcl-2 m RNA and protein at the 8th week was higher than that of immediately collected Bcl-2 m RNA and protein,with significant difference between the two(P<0.05).At the 4th week of sampling,the expression levels of Bcl-2 m RNA and protein in I/R+ Entresto group were significantly higher than those in the immedia te sampling group,which were significantly lower than those in the 8th week,with statistical significance(P < 0.05).In addition,the expression levels of Bax and Caspase-3 m RNA and protein in the I/R+ Entresto group at week 4 were lower than those in the immediately sampled specimens,while the expression levels of Bax and Caspase-3 m RNA and protein were significantly higher than those in the samples at week 8.There was statistical significance(P < 0.05).(2)Comparison between groups:Compared with Sham group,Bcl-2 m RNA and protein in I/R group were significantly decreased at all stages,and the difference was statistically significant(P<0.05).Bax and Caspase-3 m RNA and protein in I/R group were significantly increased at each stage,and the differences were statistically significant(P<0.05).At week 8,there were no significant differences in the m RNA and protein e xpression levels of Bcl-2,Bax and Caspase-3 between Sham group and I/R+ En tresto group(P>0.05).The expression levels of Bcl-2 m RNA and protein in the I/R+ Entresto group were significantly lower than those in the Sham group at immediate specimens and at week 4,and the difference was statistically significant(P < 0.05).The expression levels of Bax and Caspase-3 m RNA and protein in I/R+ Entresto group were significantly higher than those in sham group at immediate specimens and week 4,with statistical significance(P < 0.05).The m RNA and protein expression levels of Bcl-2,Bax and Caspase-3 in immediate samples of I/R+ Entresto group were not significantly different from those in immediate samples of I/R group(P > 0.05).The expression levels of Bcl-2 m RNA and protein in I/R group at week 4 and week 8 were lower than those in I/R+Entresto group at week 4 and week 8,respectively,and there were significant differences according to statistical analysis(P < 0.05).The expression levels of Bax and Caspase-3 m RNA and protein in I/R group at week 4 and week 8 wer e significantly higher than those in I/R+ Entresto group at week 4 and week 8,and there were significant differences by statistical analysis(P < 0.05).Conclusion The application of Shacobactril/valsartan effectively inhibited cardiomyocyte apoptosis,suggesting that Shacobactril/valsartan had a significant ameliorating effect on myocardial ischemia-reperfusion injury in rats.The mechanism of this effect may be achieved by regulating the Bcl-2/Bax/Caspase-3 pathway,and Shacobactril/valsartan can inhibit the expression of Bax and Caspase-3 and promote the expression of Bcl-2,which can effectively protect cardiomyocytes from injury. |
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