| Objective: To investigate the reconstruction capacity of rabbit bone marrow-derived mesenchymal stem cells (MSCs) Point-cell transplantation for ischemic myocyte, at the same time to make a comparative research on cardiomyocyte apoptosis in infarct area between rabbit bone marrow mesenchymal stem cells (MSCs) transplantation Point of zero-day and seven days. To determine the number of cell apoptosis by TUNEL, and to observe the expression of Fas, FasL, Bcl-2 protein and mRNA in myocardial cells by immunohistochemistry SABC methods, RT-PCR and Western blot.Methods: (1) The Japan rabbits that have been established model of myocardial infarction were randomly divided into 3 groups: the model group, the group with point transplantation of stem cell (0 day) , the group with point transplantation of stem cell (7 days). By the method of thoracotomy with anterior descending coronary artery ligation to establish acute myocardial infarction model, the infarction rabbits were injected with the same volume of separated, cultured, amplified, marked MSCs suspension apartly on 0 day and 7th day. In the five weeks after infarction, immunohistochemical staining Brdu is carried out in myocardial infarction area in rabbits of each group respectively to observe whether the migration of MSCs transplantats to the infarct area. The positive cells in Brdu staining were stained again by cardiac-specific troponin T in order to observe whether the migration of cells differentiate and the nature of differentiation. (2) With the method of TUNEL to determine the number of apoptosis cells, and with the method of SABC, RT-PCR and Western blot to observe the myocardial cells expressing in Fas, FasL, Bcl-2 protein and mRNA. Results: (1) There were Brdu positive cells and cTnT positive protein in Point injection MSCs group in infarct area. (2) Immunohistochemistry showed: The expression of apoptotic cells in infracted area in model group were more than normal group, the point of stem cell transplantation group(0 day)and the point of stem cell transplantation group (7 days) (P<0. 05), the expression of apoptotic cells in the point of stem cell transplantation group(7 days) was less than the point of stem cell transplantation group(0 day) (but no significant difference P>0. 05); the expression of Fas protein:the model group was more than the normal group, the point of stem cell transplantation group(0 day) and the point of stem cell transplantation group(7 days), the point of stem cell transplantation group(7 days) was less than the point of stem cell transplantation group(0 day) ; the expression of FasL protein: the point of stem cell transplantation group(0 day)more than the normal group, the model group and the point of stem cell transplantation group(7 days) (P<0. 05) .The expression of Bcl-2 protein: the model group was less than the normal group, the point of stem cell transplantation group(0 day) and the point of stem cell transplantation group(7 days)(P<0. 05) ,the point of stem cell transplantation group(7 days)more than the point of stem cell transplantation group(0 day) (but no significant difference P>0. 05). (3) RT-PCR and Western Blotting showed: Compared with the normal group, Fas mRNA levels: the model group and the two groups with point injection of stem cells was significantly increased (P <0. 05); FasL mRNA levels: the model group and the two groups with point injection of stem cells was significantly increased (P <0.05); Bcl-2 mRNA levels: Model group decreased significantly (P <0.05), the two groups with point injection of stem cells has increased, but increased in varying degrees, the group with point transplantation of stem cell (0 day) was significantly higher (P <0. 05), the group with point transplantation of stem cell(7days) also increased , but statistical significance. Compared with model group, Fas mRNA levels: the two groups with point injection of stem cells decreased significantly (P <0. 05); FasL mRNA levels: the two groups with point injection of stem cells decreased significantly (P <0. 01); Bcl-2 mRNA: the two groups with point injection of stem cells increased significantly (P <0. 05) Compared with the group with point transplantation of stem cell(0 day), Fas mRNA levels: stem cells, the group with point transplantation of stem cell (7 days) decreased significantly (P <0. 05); FASL mRNA levels: the group with point transplantation of stem cell (7 days) decreased significantly (P <0. 05); Bcl-2 mRNA levels: the group with point transplantation of stem cell (7 days) was also significantly decreased (P <0. 05). Compared with the normal group, Fas and FasL protein levels: the two groups with point injection of stem cells was significantly increased (P <0. 05); Bcl-2 protein levels: the model group decreased significantly (P <0. 05), The two groups of acupoint Injection Stem Cells have increased but increased in varying degrees, the group with point transplantation of stem cell (0 day) was significantly higher (P <0. 05), the group with point transplantation of stem cell (7 days) was significantly higher(P <0. 05). Compared with model group, Fas protein: the two groups with point injection of stem cells decreased significantly (P <0. 05); FasL protein: the two groups with point injection of stem cells decreased significantly (P <0. 05); Bcl-2 protein: the two groups with point injection of stem cells increased significantly (P <0. 05).Compared with the group with point transplantation of stem cell (0 day), Fas protein: stem cells, the group with point transplantation of stem cell (7days) decreased significantly (P <0. 05) ; FASL protein: the group with point transplantation of stem cell (7 days) decreased significantly (P <0.05) ; Bcl-2 protein: the group with point transplantation of stem cell (7 days) was also significantly decreased (P <0.05).Conclusion: (1) Point transplantation of bone marrow mesenchymal stem cells, can moved to the infarct zone, and can differentiate into myocardial cells; (2) Point transplantation of bone marrow mesenchymal stem cells can reduce the number of cardiomyocyte apoptosis in the infracted and surrounding zone, which may be caused by the reduction of Fas, FasL and the increasing of Bcl-2 to adjustments. (3) Bone marrow-derived mesenchymal stem cells Point-cell transplantation for 7 days can reduce more apoptosis cells than bone marrow-derived mesenchymal stem cells Point-cell transplantation for 0 day.
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