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MicroRNA-24 protects against ischemic injury in rat cardiomyocytes

【摘要】:正MicroRNAs are a class of novel regulators of gene expression.The present study aimed to investigate the function of microRNA -24(miR -24) in ischemic cardiomyocytes,based on a microarray screening in which miR - 24 expression was significantly enhanced in the ischemic myocardium of intact rats.In primary cultured rat cardiomyocytes,ischemia was generated by removing serum,oxygen and glucose.MiR - 24 mimic or inhibitor was transfected to induce or suppress miR - 24 expression,and their effects on ischemic - induced injury were evaluated by measuring lactate dehydrogenase(LDH) release,cell viability,and the rates of apoptosis and necrosis.LDH release as an indicator of cell damage was measured colorimetrically in the media. Cell viability was determined by the 3 -(4,5 - dimethylthiazol -2-yl) -2,5- diphenyl tetrazonium bromide (MTT) assay.Apoptosis and necrosis were determined by Hoechst 33258 staining and Annexin V/PI dual staining with flow cytometry.The results showed that the expression level of miR -24 was increased by 1.76±0.22 and 1.93±0.03 fold in in vivo rat myocardium subjected to 4 - h ischemia and in cultured myocytes exposed to 24 - h ischemia,respectively.In ischemic cultures of cardiomyocytes,the overexpression of miR - 24 significantly decreased LDH release[(33.7±1.4) vs(42.8±4.2) U/L,P0.01],and enhanced cell viability[(68.3±5.4)%vs(53.7±9.1)%of normal control values,P0.05].Lower rates of apoptosis [(11.5±5.7)%vs(18.7±5.5)%,P0.05]and necrosis[(3.5±2.0)%vs(6.0±1.2)%,P0.05]were detected by Annexin V/PI staining.Contrary to the observations with miR -24 overexpression,the deficiency of miR -24 in ischemic myocytes resulted in the highest necrosis rate[(9.9±2.0)%,P0.01 vs ischemia]and the lowest cell viability[(39.5±1.7)%of normal control values,P0.01 vs ischemia]. MiR - 24 deficiency in normal cultured myocytes also induced significant apoptosis and decrease in cell viability. Additionally,the expression of the pro - apoptotic gene BCL2L11 was found to be down - regulated by miR -24.In conclusion,this study showed that miR -24 was upregulated and protective against myocardial ischemia by inhibiting BCL2L11.It may represent a potential novel treatment for ischemic heart disease.

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