Screening And Cloning The Genes Relared To Rats After Transplantation Bone Marrow Mesenchymai Stem Cells In Cardiomyopathy

Background:Cardiovascular diseases are the most frequent cause of death in the western hemisphere. Cell transplantation has been demonstrated to be of therapeutic value for the repair of damaged myocardium in animal models,which will be an ideal seed cell. Use of bone marrow mesenchymal stem cell (BMMSCs)has clinical advantages,because these cells have characters of originating conveniently, expanding easily,and no immunity. BMMSCs can differentiate into myocytes and improve heart function in vivo and vitro.Our earlier studies have confirmed BMMSCs can transdifferentiate into myocytes and improve heart function in 2 weeks.But the mechanism is not clear now. So we hypothesize cell transplantation will cause genetic change,such as some genes opening and closing and the opened genes changing in quantity. Objective :To study BMMSCs improved heart function and preliminary study the machanism of improvement heart function with suppression subtractive hybridization(SSH). Methods :BMMSCs were isolated from thighone and tibia of Wistar rats ,purified by adheresion-screening method, and expanded in vitro .Intraperitioneal injection of doxorubicin established cardiomyopathy models.(1~5)×106 BMMSCs were injected into left ventricular myocardium of rats with cardiomyopathy rats.TTE technique detected heart function after operation. Thighbone bone marrow was aspiratedand stromal cells were cultured and transplanted into cardiomyocytes.1 weeks later,screening the differential genes between tester (cardiomyopathy rats injected BMMSCs) and driver(cardiomyopathy rats injected equivalent volume culture medium)with suppression subtractive hybridization. Results : After 8 weeks of intrapertoneal injection of doxorubicin,LVEF LVFS decreased 20-30% compared with normal group,which are cardiomyopathy model rats. LVEF LVFS are no significant difference between cardiomyopathy model rats and cardiomyopathy model rats before transplantation.However,there is significant difference of LVEF LVFS after transplantation BMMCSs in 2 weeks,which can improve the heart function,and among the 17obtained positive clone,13 were highly expressed and 4 lowly. Conclusion: The heart function has significant improvement after transplantation BMMSCs. The mechanisms of transdifferentiating into cardiacmyocytes and improvement heart function may be the differential expressed genes which are mitochondrial synthesized genes and protein of contracting synthesized genes .