The Effect Of BMP9 In Cpcs Ameliorate The Heart Function In The Rat Model Of Myocardial Infarction

PART ONE : AMPLIFICATION OF AdBMP9 AND AdGFP, THE INFECTION OF CARDIAC PROGENTIOR CELLSObjective: Amplifying the AdGFP and AdBMP9 with 293 cells to get high-titer adenovirus; Infect the CPCs with AdBMP9 and improve the expression of BMP9 of the CPCs.Methord: Cultivate the 293 cells and amplify the adenovirus with 293 cells. Cultivate the cardiac progenitor cells. CPCs were infected by AdBMP9 which express green fluorescent protein. After twenty-four hours, the expression of BMP9 in CPCs were tested by semi-quantitative RT-PCR.Result:1. We got high titer adenovirus by amplifying the AdGFP and AdBMP9 with 293 cells.2. After CPCs infected with adenovirus 24 hours, the efficiency of AdGFP and AdBMP9 is 45% and 46%. 3. In three groups, the expression of BMP9 is improved dramatically in CPCs infected with AdBMP9.Conclusion : The titer of AdBMP9 and AdGFP is improved though 293 amplification. The expression of BMP9 is improved dramatically in CPCs infected with AdBMP9. PART TWO: THE STUDY OF BMP9 IN CPCs AMELIORATE HEART FUNCTION IN THE RAT MODEL OF MYOCARDIAL INFARCTIONObjective: To test whether cardiac progenitor cells (CPCs) which induced by bone morphogenetic proteins9 (BMP9) could improve the heart function or not.Methord: Twenty six male rats were divided into three groups. MI group (n=10) ,MI+GFP group (n=8), MI+BMP9 group (n=8.). Four weeks after transplantion, the change of CPCs morphology was tested though the expression of cardiac-specific structural proteins alpha myosin heavy chain(a-MHC) and a-actin of the implanted CPCs with HE staining and immunofluorescent analysis. The myocardium infarct area was detected though Masson staining. Heart function was evaluated by echocardiography, in preoperative and postoperative four weeks.Result:1.MI models were established successfully by blocking left anterior descending coronary arteries.2. Four weeks after transplantation , the heart function of the rats were compared during the three groups. In LVESD and FS level , MI+BMP9 group better than other group(p<0.05). The level of LVESD and FS in MI+GFP group was improved compared with the MI group. 3. Four weeks after transplantation, the implanted CPCs reduced the size of infracted obviously(MI+GFP VS MI ,P﹤0.05)and could be promoted by BMP9(MI+BMP9 VS MI P﹤0.01)4. The result of HE staining and immunofluorescent showed that the grafted CPCs could fuse with the normal cardiomyocytes and expressed the cardiac-specific proteins a-MHC and a-actin, at four weeks after transplantation. But the expression of cardiac-specific structural proteins is lower than the abnormal cardiac tissue. The expression of a-MHC and a-actin in MI+BMP9 group is more than MI+GFP group.( P﹤0.05) Conclusion:BMP9 could improve the expression of cardiac-specific structural proteins of CPCs and promote cardiac progenitor cells ameliorate the heart function in the rat model of myocardial infarction.