Paracrine effects of bone marrow mesenchymal stem cells on the apoptosis of adriamycin-injured cardiomyocytes:How to verify the inhibitory effects?
10.3969/j.issn.1673-8225.2010.10.009
- VernacularTitle:骨髓间充质干细胞旁分泌途径与阿霉素损伤心肌细胞凋亡:怎样证明其抑制效应?
- Author:
Junfang GUO
;
Yingyu ZHANG
;
Rong CHEN
;
Yanfang ZHOU
;
Hao WANG
;
Guohui ZHANG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2010;14(10):1755-1759
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:Several studies have shown the improvement of heart function through the introduction of mesenchymal stem cells(MSCs)from bone marrow,which may be attributed to secretion of various cytokines that accelerate endogenous reparative process,but not regeneration of cardiomyocytes.OBJECTIVE:To observe the anti-apoptotic effects of MSCs on adriamycin(ADR)-injured cardiomyocytes apoptosis in vitro through paracrine pathway.METHODS:In vitro cultured neonatal rat cardiomyocytes(3×10~8/L)were incubated into a 6-well plate,3 mL/well.72 hours later,these cells were assigned into 3 groups.The primary cultured neonatal rat cardiomyocytes in the ADR-injured and coculture groups were exposed to 1 mg/L ADR for 4 hours to establish experimental models of toxic cardiomyocytes.The normal control group was left intact.In the coculture group,rat bone marrow MSCs(BMSCs)at passage 3 were regulated to 3×10~8/L,3 mL/well was added into Millicell device for 24 hours.Following model induction,the Millicell device was inseted into above-mentioned 6-well plate to establish coculture system.The levels of cytokines were measured in the conditioned medium from three cardiomyocytes groups.Effects of BMSCs on Caspase-9 and Caspase-3 activities,apoptosis and Bcl-2 and Bax protein expression in ADR-injured cardiomyocytes were measured.RESULTS AND CONCLUSION:Compared with the normal control group.the level of cytokine including insulin-like growth factor (IGF-1)and hepatocyte growth factor(HGF)was significantly higher in the medium from ADR-injured group.the activity of caspase-9,caspase-3 and the apoptosis rate increased significantly,the expression of Bax protein was higher and Bcf-2 Protein was lower in ADR-injured group(P ＜ 0.05).Compared with the ADR-injured group,the level of IGF-1 and HGF in co-cultured group increased significantly,the apoptosis rate,Caspase-3 and Caspase-9 activities decreased significantly,the expression of Bax protein was lower while Bcl-2 Protein was higher than ADR-injured group(P ＜ 0.05).Results indicated that BMSCs show increased cytokine secretion and significant anti-apoptotic effects on ADR-injured cardiomyocytes through paracrine pathway.
