Sarcopenia-derived exosomal micro-RNA 16-5p exerts the cardio-repair disturbance via pro-apoptotic mechanism in myocardial infarction of mice

Abstract

Abstract Background Sarcopenia is a pathophysiological malfunction induced by skeletal muscle atrophy, and several studies reported an association between sarcopenia-induced cardiac cachexia and poor prognosis in heart disease. Since only a few established animal models are recently available, the underlying mechanism of disturbed cardiac repair accompanied with sarcopenia remains poorly understood. Purpose We hypothesized that specific microRNAs in sarcopenia-derived exosomes play crucial roles in disturbed cardiac repair with sarcopenia, and these microRNAs directly exacerbate cardiomyocyte injury following cardiac ischemia and reperfusion. Methods We developed a novel sarcopenia-induced cardiac repair disturbance mouse model that is induced by tail suspension (TS) 7 days after a 45-min coronary occlusion of cardiac ischemia and reperfusion (I/R). The reduction of the left ventricular ejection fraction (LVEF) after I/R was compared in mice with TS [I/R-TS(+), n=14] and without [I/R-TS(−), n=12] by echocardiography. To investigate the exosomal mechanism of cardiac repair disturbance, a comprehensive analysis of extracted exosomal microRNAs from mice serum was performed in the 2 groups at day 8. Then, we investigated the impact of the identified candidate microRNA in neonatal rat cardiomyocytes (NRVMs). After 4 days in primary culture, candidate microRNA was transfected into NRVMs under hypoxic culture conditions. TUNEL analysis and quantitative PCR analysis of apoptosis-related genes were performed on the NRVMs. Results At day 8 after I/R, the LVEF of I/R-TS(+) was not significantly ameliorated compared to that of I/R-TS(−) (ΔLVEF; 1.59±6.92 vs. 8.04±7.71% p=0.034). Four candidate microRNAs obtained from I/R mice serum were identified in the microRNA array analysis. The re-analysis of these candidate micro-RNAs using all I/R mice demonstrated that the level of mir-16-5p in I/R-TS(+) was raised by approximately nine-fold than that in I/R-TS(−) (9.67±13.35 vs. 0.99±1.41, p<0.05). Next, an in vitro experimental model using a microRNA mimic revealed that apoptosis in NRVMs was greatly enhanced by the transfection of a mir-16-5p mimic in hypoxic culture conditions (mir-16-5p vs. control = 5.77±2.84 vs. 1.72±0.55%, p<0.01). Furthermore, by qRT-PCR analysis, the expression of CASP3 and TRP53 were upregulated in NRVMs treated with a mir-16-5p mimic than in control NRVMs. Conclusion Myocardial I/R injury in sarcopenia ended in cardiac repair disturbance accompanying with the enhanced expression of exosomal-mir-16-5p. A pro-apoptotic effect of mir-16-5p may exacerbate myocardial I/R injury and thus can be a novel therapeutic target for cardiac repair disturbance in sarcopenia. Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): Japan Society for the Promotion of Science