TMeglumine Cyclic Adenosine-induced Differentiation Of Bone Marrow Mesenchymal Stem Cells Into Cardiomyogenic Cells In Experimental Research On And Analysis Of Curative Effect In Patients With Diastolic Heart Failure

First part of meglumine adenosine cyclophosphate on differentiation of bone marrow mesenchymal stem cells into cardiomyogenic cells in study on effects and mechanismsObjective: Explore the Meglumine Cyclic Adenylate(MCA) on differentiation of bone marrow mesenchymal stem cells into cardiomyogenic cells effect and mechanism.Methods: Cultured by attachment method of bone marrow mesenchymal stem cells from bone marrow.Flow cytometry identification of cell surface antigens, CD44, CD90, CD45 expression. 1)Take the vigorous growth of the P3 cell divides into 3 groups: Normal control group, the MCA group, 5-AZA Cytidine(5-Aza) group, Cell culture of BMSCs measured by fluorescence quantitative RT-PCR after 4 weeks cardiac-specific genes Gata-4m RNA, Cx43 m RNA expressions, detection of cardiac specific Western-blotting protein c Tn T and Cx43 m RNA. 2)3rd generation cells are divided into 3 groups: control group, MCA group, MCA+LY294002 group, train until 4 weeks after the detection of cell pathway protein Western-blotting PI3 K, p-AKT, p-Foxo3 a, DOK5 expression.Results: Through the whole bone marrow adherent culture of BMSCs in vitro by flow cytometry after identification, cell surface antigen consistent with basic characteristics of BMSCs logo.1)1x10-3mol/L MCA-induced 3 days after the intervention GATA-4m RNA,Cx43 m RNA the relative expression was highest(P<0.01), cardiac muscle specific proteins c Tn T and Cx43 expression was significantly higher than the normal control group, 5-Aza Group(P<0.01). 2)the MCA group PI3 K, p-AKT, p-Foxo3 a, and the relative expression of DOK5 was significantly higher than the normal control group, MCA+LY294002 Group(P<0.01).Conclusion: MCA(1x10-3mol/L)for 3 days may induce differentiation with best results.The mechanism may be related to PI3K/AKT cell signaling pathways.Second part of meglumine cyclic Adenylate in treatment of patients with diastolic heart failure analysisObjective: To investigate the therapeutic effect and possible mechanism of MCA in patients with diastolic heart failure(DHF).Methods: 1) According to the 2012 ESC heart failure guidelines through clinical symptoms, and echocardiography examination diagnosed DHF patients in 66 cases, including 31 cases of patients with conventional drug treatment, such as beta blockers, ACEI or ARB, diuretic, as control group; 35 cases of patients in addition to the application of conventional drug treatment and injection with the treatment of MCA, set as the experimental group.2) The peripheral blood of the control group and the experimental group were collected for 24 hours and 2 days after treatment for one cycle(fifteenth days after treatment). The value of plasma NT-Pro BNP was measured by 1 tube, and the changes of cardiac function before and after treatment in two groups were analyzed. 1) the other 3 blood cells were separated and extracted by human lymphocyte separation to extract a single nuclear cell, and the total RNA was extracted. The relative expression of GATA-4 and Cx43 genes in the two groups were measured before and after treatment by RT-PCR method.Results: 1) 66 patients were actively cooperate with the treatment, the treatment process is not a case of exit, no death. 2) Two groups of patients admitted to detection of NT pro BNP numerical concentration had no significant difference, in the experimental group and the control group before and after treatment with numerical concentration of NT pro BNP were improved, the experimental group after treatment, numerical concentration of NT pro BNP significantly decreased, degree of decrease of serum NT pro BNP and the experimental group and the control group treatment after, the difference is statistically significant(P<0.01), MCA can improve DHF effect.3) Two groups of patients admitted to detect cardiac specific gene GATA-4 m RNA and Cx43 m RNA expression had no significant difference (P>0.05),after treatment in the experimental group than in the control treatment group after GATA-4 m RNA and Cx43 m RNA relative expression was significantly increased(P<0.01),and compared in the experimental group and the control group before and after treatment of GATA-4 m RNA and Cx43 m RNA in the rising degree, the difference is statistically significant(P<0.01),.Conclusion: MCA can improve the therapeutic efficacy of DHF patients, the mechanism may be related to MCA can induce BMSCs in peripheral blood to differentiate into cardiomyocyte like cells.