Cellular FLICE-like inhibitory protein alleviates myocardial ischemia/reperfusion injury via inhibiting necroptosis / 中华急诊医学杂志

ABSTRACT

Objective:To explore the regulatory effect of cellular FLICE-like inhibitory protein (cFLIP) on myocardial ischemia-reperfusion injury based on the RIPK1/RIPK3/MLKL-mediated necroptosis pathway.Methods:The cardiomyocyte hypoxia/reoxygenation (H/R) model was constructed by hypoxia for 4 h/reoxygenation for 12 h, and the rat ischemia reperfusion (I/R) model was constructed by ligating the left anterior descending artery for 30 min and reperfusion for 3 h. CCK-8 method was used to detect the viability of cardiomyocytes in each group. DAPI/PI double staining was used to observe changes in necrosis rate of myocardial cell. STRING database was used to predict the protein interaction network of cFLIP. TTC staining was used to detect the area of myocardial infarction in each group of rats, and the protein expression of cFLIPL, cFLIPS, p-RIPK1, p-RIPK3 and p-MLKL were detected by Western blot.Results:In cardiomyocyte H/R injury and myocardial tissue I/R injury, the protein expressions of cFLIPL and cFLIPS were significantly down-regulated, while the levels of p-RIPK1, p-RIPK3 and p-MLKL were increased significantly. Up-regulating the protein expression of cFLIPL and cFLIPS could significantly reduce the damage of cardiomyocytes and the rate of cell necrosis induced by H/R, and decrease the area of myocardial infarction caused by I/R. STRING database results showed that cFLIP had direct protein interactions with RIPK1 and RIPK3. Overexpression of cFLIP in cardiomyocyte and myocardial tissue significantly inhibited H/R or I/R induced the phosphorylation levels of RIPK1, RIPK3 and MLKL.Conclusions:Overexpression of cFLIP can significantly inhibit the RIPK1/RIPK3/MLKL-mediated necroptosis, thereby reducing myocardial cell damage and decreasing the area of myocardial infarction.