The Function Of Different Passages Bone Marrow-derived Mesenchymal Stem Cells On The Treatment Of Acute Myocardial Infarction

Objective: To research the function of different passages bone marrow mesenchymalstem cells on the treatment of acute myocardial infarction；Compare the different passagesbone marrow-derived mesenchymal stem cells in to improve cardiac function aftermyocardial infarction;Compare different passages of bone marrow stem cells distributionin vivo; Detect cytokines secreted by stem cells in ischemic hypoxic conditions；Analysisof the changes of cytokines with the generation time；Study effect of paracrine mechanismson the treatment of acute myocardial infarctionMethord: Stem cell culture and supernatant extract:bone marrow were isolated from8weeks old SD rat’s femur and tibia under sterile conditions. Bone marrow mesenchymalstem cells were extracted and cultured by the differences adherent of whole bonemarrow.The3rd generation and6th generation of stem cells were marked by XenoLightDiR whose final concentration was320μg/mL, at the same time the extraction of the3thgeneration and the6th generation of stem cell culture supernatant were finished. Model:we ligated left anterior descending coronary artery to established models of acutemyocardial infarction. Experimental groups: SD rats were25, either male or female, wererandomly divided into5groups，48hours after myocardial infarction model estabilshed,groups were injected via tail vein， group A: the3rd culture supernatant1ml; Group B: the6th culture supernatant1ml; group C: the3rd MSC5*106/1ml; Group D:6th MSC5*106/1ml; group E: control normal saline1ml.Cardiac functions of each group weredetected by echocardiographic prior myocardial infarction,after myocardial infarction,aftertransplantation.ELISA was applied to the analysis cytokines in the serum.Small animalvivo imaging and enhanced CT were applied to observate stem cells distribution in vivo and the changes about myocardial infarction.Result:(1) Doppler ultrasound echocardiography in detection of cardiac functionbefore and after myocardial infarction and after transplantion of stem cell and culturesupernatant, that found the6th generation MSC improvement of cardiac function betterthan the3rd generation of the MSC;(2) Whole body imaging of small animals observed inthe6th generation MSC of the fluorescence intensity is higher than3rd generation of theMSC;(3) Protein analysis technology to detect cytokine levels indifferent passages of stemcell culture supernatant,6th generation MSC culture supernatant MCP-1and TIMP-1levels lower than the3rd generation of MSC culture supernatant, LIX content higher thanthe3rd generation of MSC culture supernatant;(4) ELISA method detected the higherlevel of MCP-1, TIMP-1and LIX in vivo after transplantion of stem cell and culturesupernatant, the increased of the amount of MCP-1and TIMP-1in the6th generationMSC group are less than the3rd generation MSC group, the increased amount of LIXhigher than the3rd generation MSC group.Conclusion:The6th generation MSC improvement of cardiac function are better than the3rdgeneration of the MSC after myocardial infarction，related to reduce proinflammatorycytokine secretion, increase in the angiogenic cytokine secretion.