Effect of CrkL silence on hypoxia/reoxygenation-induced apoptosis and survival inhibition in cardiomyocytes and the underlying mechanisms / 第二军医大学学报

ABSTRACT

Objective: To investigate the effect of CrkL silence on hypoxia/reoxygenation (H/R)-induced apoptosis and survival inhibition in cardiomyocytes and the underlying mechanisms. Methods: H9C2 cardiomyocytes were infected with blank, negative lentivirus and CrkL RNAi lentivirus (CrkL mRNA silence), and then treated with H/R separately. The cardiomyocytes were divided into blank group, negative lentivirus group, CrkL silence group, blank + H/R group, negative lentivirus + H/R group and CrkL silence + H/R group. The expression of CrkL mRNA was detected by RT-PCR, and the expression of CrkL and p-ERK1/2 protein was detected by Western blotting analysis. The apoptosis rate of cardiomyocytes was analyzed by flow cytometry, and the cell proliferation rate was analyzed by MTT. Results: Compared with blank and negative lentivirus groups, CrkL silence group had significantly decreased CrkL mRNA and protein, p-ERK1 protein, p-ERK1/2 protein and proliferation rate (P<0.05 or P<0.01), and significantly increased apoptosis rate (P<0.01). The same was true for CrkL silence + H/R group when compared with the blank + H/R and negative lentivirus + H/R groups. Conclusion: CrkL silence can aggravate H/R-induced apoptosis and survival inhibition in cardiomyocytes, which might be mediated by the decrease of p-ERK1/2 protein expression.