Objective To investigate the effects of Irisin on oxidative stress and apoptosis of cardiomyocytes under simulated microgravity.Methods H9C2 cells were cultured under simulated microgravity by 2D clinostat and were randomly divided into 4 groups:control group(CON group),Irisin-treated group(Irisin group),simulated microgravity group(SMG group),simulated microgravity+Irisin-treated group(SMG+Irisin group).qRT-PCR was used to detect mRNA expression levels of fibronectin type Ⅲ domain-containing protein 5(FNDC5).ELISA was used to determine the content of Irisin in cell supernatant.Western blotting was used to detect the expression levels of FNDC5,Irisin,and proteins related to oxidative stress and apoptosis.Flow cytometry was used to detect cell apoptosis rate and reactive oxygen species(ROS)levels.The levels of malondialdehyde(MDA)and glutathione(GSH)and the activity of superoxide dismutase(SOD)were determined by detection kits.Results In vitro experiments showed that compared with the CON group,the expressions of FNDC5 and Irisin decreased in H9C2 cells of SMG group,and the content of Irisin reduced in cell supernatant(all P＜0.01),suggesting a decreased Irisin production in H9C2 cells after simulated microgravity.After 72 h of simulated microgravity,ROS production and MDA content of H9C2 cells were increased,GSH content decreased(all P＜0.01),SOD activity decreased(P＜0.01),and SOD1 and SOD2 protein expressions decreased(both P＜0.01),indicating an increased oxidative stress level of H9C2 cells after simulated microgravity.Furthermore,compared with the CON group,the cell apoptosis rate and the protein expression of cleaved caspase-3 increased,the expression of anti-apoptotic protein Bcl-2 decreased(all P＜0.01),and the expression of pro-apoptotic protein BAX increased(P＜0.05)in the SMG group,revealing an increased apoptosis of H9C2 cells after simulated microgravity.Compared with the SMG group,ROS levels and MDA content were decreased(both P＜0.01),GSH content,SOD activity,and SOD1 and SOD2 protein expressions were significantly increased(all P＜0.01),cell apoptosis rate was decreased(P＜0.01),the expressions of BAX and cleaved caspase-3 were significantly decreased(both P＜0.01),and the expression of Bcl-2 was significantly increased(P＜0.01)in the SMG+Irisin group.Conclusion Irisin can alleviate the damage of cardiomyocytes caused by simulated microgravity by reducing oxidative stress levels and inhibiting the apoptosis of cardiomyocytes,which has the potential to be a candidate drug for anti-myocardial injury.
关键词
鸢尾素/模拟失重/氧化应激/心肌细胞凋亡
Key words
Irisin/simulated microgravity/oxidative stress/apoptosis of cardiomyocytes
维普
万方数据