Effects Of Sofren Injection On Mitochondrial Function And Mitochondrial Dynamics Of Hypoxia/Reoxygenation Cardiomyocytes

Objective:To establish a rat model of hypoxia/reoxygenation(H/R)injury in rat primary cardiomyocytes,and to investigate the effects of Sofren injection on mitochondrial function and mitochondrial dynamics-related protein expression,clarify its role in cardiomyocyte H/R injury and its possible mechanism.Methods:Neonatal SD rat primary cardiomyocytes were isolated and cultured.The cardiomyocytes were well divided into five groups:①normal control group,②H/R group,③post-intervention group(H/R group+Sofren injection),④pre-intervention group(Sofren injection+H/R group),⑤pre and post-intervention group(Sofren injection+H/R group+Sofren injection).MTT colorimetric method was used to determine the effect of Sofren injection on the activity of cardiomyocytes,and the optimal concentration was determined.The levels of mitochondrial function,reactive oxygen species(ROS),adenosine triphosphate(ATP)and mitochondrial membrane potential(MMP)were measured by multifunctional microplate reader.Western blotting was used to detect the expression of OPA1,Drp1 and p-Drp1 in mitochondrial dynamics-related proteins.Results:1.Effects of Sofren injection on ATP and ROS levels in primary cardiomyocytes injured by H/R:Compared with the normal control group,the level of myocardial ATP was decreased in the H/R group,and ROS was increased(P<0.05).Compared with the H/R group,the level of myocardial ATP was increased and ROS was decreased in post-intervention group,pre-intervention group and pre and post-intervention group(P<0.05).The level of ATP in the cardiomyocytes increased and the level of ROS decreased in pre and post-intervention group,and the difference was statistically significant(P<0.05)compared with the pre-intervention group and the post-intervention group.There was no significant difference in ATP and ROS levels between the pre-intervention group and the post-intervention group(P>0.05).2.Effects of Sofren injection on MMP in primary cardiomyocytes injured by H/R:Compared with the normal group,the level of MMP was decreased in the H/R group(P<0.05).Compared with the H/R group,the level of MMP was increased in the post-intervention group,pre-intervention group and pre and post-intervention group(P<0.05).The level of MMP in the cardiomyocytes increased in pre and post-intervention group,and the difference was statistically significant(P<0.05)compared with the pre-intervention group and the post-intervention group.There was no significant difference in MMP levels between the pre-intervention group and the post-intervention group(P>0.05).3.Effects of Sofren injection on the expression of OPA1 in primary cardiomyocytes injured by H/R:Compared with the normal group,the level of OPA1 was decreased in the H/R group(P<0.05).Compared with the H/R group,the level of OPA1 was increased in post-intervention group,pre-intervention group and pre and post-intervention group(P<0.05).There was no significant difference in OPA1 expression between thepost-intervention group,pre-intervention group and pre and post-intervention group(P>0.05).4.Effects of Sofren injection on the expression of Drpl and p-Drpl in primary cardiomyocytes injured by H/R:The expression of Drpl was unchanged among groups(P>0.05),and the expression of p-Drpl was different.Compared with the normal group,the level of p-Drp1 was increased in the H/R group(P<0.05).Compared with the H/R group,the level of p-Drpl was decreased in post-intervention group,pre-intervention group and pre and post-intervention group(P<0.05).The level of p-DRP1 in the cardiomyocytes decreased in pre and post-intervention group,and the difference was statistically significant(P<0.05)compared with the pre-intervention group and the post-intervention group.There was no significant difference in p-Drpl levels between the pre-intervention group and the post-intervention group(P>0.05).Conclusion:Sofren injection has a significant protective effect on injured cardiomyocytes by H/R.The mechanism may be to reduce mitochondrial dysfunction by inhibiting the imbalance of mitochondrial dynamics in damaged cardiomyocytes,thereby reducing myocardial ischemia-reperfusion injury(MIRI)and exerting a certain myocardial protection.