Effect Of Danshen Injection Combined With Cardiac Stem Cell Transplantation On Acute Myocardial Infarction In Mice

Aim:In this study, we explore the therapeutic potential of danshen injection in cardiac stem cell (CSC) transplant after cardiac infarction by in vivo and in vitro tests. Method:(1) Annexin V-PI, Caspase-3and CCK-8were detected to assess hypoxia induced myocytes apoptosis. We observed whether danshen injection could prevent myocytes from apoptosis in hypoxia.(2) Sca-1positive CSC was cultured in this study. Anti-apoptosis and pro-proliferation effect of danshen injection on CSC were detected via protein level of caspase-3and CCK-8, as well as Edu kit respectively.(3) In vivo, we detected whether danshen injection in different dose could increase Sca-1positive CSC and VEGF, IL-6as well as BAD expression in myocardium.(4) We combined CSC and danshen injection in experimental MI mice to verify whether danshen injection could enhance CSC transplantation effect on MI. In histology analysis, angiogenesis, tissue fibrosis, myocyte apoptosis and Akt expression were detected. Result:(1) In hypoxia, high dose danshen injection could significantly reduce myocytes apoptosis via flow cytometry. Furthermore, decreased caspase-3and BAD and increased FGF and Akt expression were observed in danshen injection treated myocytes.(2) Danshen injection could significantly protect CSC against hypoxia induced apoptosis via up-regulated caspase-3and down-regulated CCK-8activity. On the other hand, CSC proliferation was increased in danshen injection treated group.(3) After treated by high dose danshen injection one week, the sca-1positive CSCs were significantly increased in myocardium. VEGF and IL-6expression were also increased in high dose Danshen injection treated mice, meanwhile BAD expression was decreased. No significantly difference of sca-1positive CSCs, VEGF. IL-6and BAD expression were observed between low dose Danshen injection group and control group.(4) Danshen injection could improve CSC transplantation effect in experimental mice MI model. Increased angiogenesis and proliferation cell were observed in CSC and danshen injection treated group, compared with CSC treated and control group, meanwhile, decreased apoptosis and tissue fibrosis were also observed. Furthermore, Akt expression and phosphrylation were both increased in CSC and danshen injection treated group. Conclusion:Combined danshen injection and CSC transplantation could improve CSC survival after transplantation, induce tissue angiogenesis and inhibit cell apoptosis and tissue fibrosis. All these therapeutic effects contributed to reserve heart function and prevent cardiac remodeling after MI.