The Effect Of 3-nitro-N-methyl Salicylamide On The Preservation Of Isolated Rat Hearts

Objective: Cold crystalloid preservation is still the standard method for donor heart preservation before heart transplantation.The key point for the success of heart transplantation is isolated heart preservation time and myocardial ischemia reperfusion injury after transplantation.There are many kinds of heart preservation solution researched to make a better preservation result,but nearly all of them can't maintain the heart function at longer ischemia preservation state.Therefore,we made a new heart preservation solution characterized by 3-NNMS,which has a half inhibitory effect for mitochondria complex?,and also by putting it into the clinical preservation solution to make a better heart preservation effect at longer ischemia preservation.We speculate that this agent can make it through reducing mitochondria ROS via slowing down mitochondria electron transport rate,therefore,mitochondria injury can be reduced by less oxidative stress and obtain a better cell function.Hence,a new heart preservation solution research idea was wish to verified through our research,and make a theory base for other research and clinical apply.Methods: 49 male Wistar rats was randamly divided into 7 groups(7 per group): a: control group,b:Base group,the new cardiaplegia solution without NNMS,c:NNMS group,the new cardiaplegia solution with 3-NNMS,d:HTK group,e:Celsior group,f:HN group,HTK solution+3-NNMS,g:LN group,Celsior solution + 3-NNMS.Isolated hearts were perfused 15 minutes to obtain a balanced state and then were immediately sotraged at 4? for 8 hours after arrested and immerged with different cardioplegia solutions.The effects of 3-NNMS on myocardium function were assessed by analyzing the hemodynamics,myocardium/perfusate CK-MB/LDH /c Tn-T,as well as HE staining,and immunohistochemistry for MPO.For mitochondria viability and function,we tested mitochondria RCR/MMP/ ATP/ROS.Besides,we also demonstrated the safety of NNMS on H9 CM cell culture via MTT and seahorse cell respiration test.Results: The use of 3-NNMS immproved the heart rate and effectively decreased the relase of enzymes during reperfusion and therefore reduced the myocardial injury.Mitochondrial membrane potential was effectively increased in 3-NNMS group compared with the others,and mitochondrial SOD activity was significantly higher than others.Morever,the supplement of 3-NNMS in HTK and Celsior both have a better antioxidant ability and relatively lower concentration of myocardial enzymes.Cell tests also show no difference on cellular activity and respiration capacity analysis under series 3-NNMS concentration.Conclusion: The new 3-NNMS cardioplegic solution make a nearly or better protective effect on isolated hearts function compared with HTK and Celsior.And,3-NNMS decreased the inflammation effectively.Moreover,the supplementation of 3-NNMS to HTK and Celsior could significantly improve its cardioprotective effects.Besides,3-NNMS is completely safe for H9C2 cell.Hence,3-NNMS is a potential reagent in myocardioprotective research during ischemia/reperfusion injury.