Protective Effects Of St.Thomas Solution Supplemented With Salvia Miltiorrhiza Extract And Fructose 1,6 Diphosphate On The Isolated Rat Heart By Continuous Hypothermic Microperfusion Method

Objective:The protective effects of St.Thomas solution supplemented with Salvia miltiorrhiza extract(SM) and fructose-1,6-diphosphate(FDP) on the isolated rat heart were investigated by continuous hypothermic microperfusion preserving method.The possible protein biomarkers that could predict the quality of preserved heart were determined through comparison of protein SDS-PAGE results with functional data of preserved hearts.Methods:Twenty-four SD rats were divided into 4 groups randomly(n=6):control group;SM group;FDP group;SM+FDP group.The isolated rat hearts were arrested with ice-cold St.Thomas solution and perfused with K-H solution for 30 min on the Langendorff model.The cardiac function(coronary flow,heart rate,left ventricular developed pressure,±dp/dt max) were determined.The hearts were arrested and preserved at 4℃for 6 h using continuous microperfusion.After preservation,cardiac functional measurements were repeated.The activities of lactate dehydrogenase(LDH) and creatine kinase(CK) in preservation fluid infusion and the water content of the heart tissue were quantified.The protein in the remanent coronary outflow fluid was concentrated by DOC-TCA precipitation method for SDS-PAGE and followed by silver staining.We attempted to seek the proteins that can predict the quality of preserved heart,and determinate molecular weight of those.The image of protein gel was treated with ImageJ software,and the relativities between the gray value of protein bands and functional data of preserved hearts were analysed.Results:During 20 min reperfusion period,in SM group,FDP group and SM+FDP group,the value of LVDP,-dp/dt max and recovery rate of +dp/dt max were significantly higher than that of control group(P＜0.05);+dp/dt max of SM group and FDP group were obviously higher than that of control group(P＜0.05).Compared with the control group,the activities of LDH in SM group,FDP group,SM+FDP group and the activities of CK in FDP group,SM+FDP group were significantly lower(P＜0.05).The water content of heart tissue in SM group,FDP group,SM+FDP group were lower than that of control group,but had no obvious difference compared with four groups(P＞0.05).Analysed the result of SDS-PAGE silver staining,three protein bands,band A(15.1kDa),band B(16.6kDa),band C(30.2kDa) were positively correlated with the activity of CK.Conclusion:Supplementation of SM or FDP to St.Thomas solution could improve the protective effects on the isolated rat hearts preserved by continuous hypothermic microperfusion.Supplementation of SM and FDP in combination did not Show any more beneficial effects over supplementation of SM or FDP alone.Through the preliminary protein analysis of the coronary outflow fluid within the preservation period,the relation between band A,B,C and myocardial damage is worth for further study.