晚期内皮祖细胞条件培养液对阿霉素诱导的心肌细胞凋亡的影响

Effect of late endothelial progenitor cells conditioned medium on cardiomyocytes apoptosis induced by adriamycin

目的观察晚期内皮祖细胞条件培养液对阿霉素诱导心肌细胞凋亡的影响。方法分离培养并鉴定内皮祖细胞和心肌细胞,制作晚期内皮祖细胞条件培养液以及阿霉素诱导的心肌细胞凋亡模型,实验分3组:对照组、模型组及实验组。Hoechest33342/PI及Annexin-FIFC/PI双染法检测各组心肌细胞凋亡率。MTT法检测心肌细胞存活率。qRT-PCR以及Western Blot法检测心肌细胞中脑钠肽(BNP)和转化生长因子-β1(TGF-β1)的mRNA及蛋白的表达。结果与对照组相比,模型组的心肌细胞凋亡率升高,与模型组相比,实验组的心肌细胞凋亡率下降,差异均有统计学意义(P <0.05)。与对照组相比,模型组的BNP的mRNA及蛋白的表达均下调,TGF-β1的mRNA及蛋白的表达上调,与模型组相比,实验组的BNP的mRNA及蛋白的表达均上调,TGF-β1的mRNA及蛋白的表达均下调,差异均有统计学意义(P <0.05)。结论晚期内皮祖细胞条件培养液抑制了阿霉素诱导的心肌细胞凋亡,其可能是通过抑制TGF-β1及上调BNP的表达所实现的。

Objective To observe the influence of late endothelial progenitor cells conditioned medium(EPCs-CM)on cardiomyocytes apoptosis induced by Adriamycin(ADR).Methods EPCs and cardiomyocytes were isolated,cultured separately and identified.Preparation of late EPCs-CM and ADR induced cardiomoycytes apoptosis model.The cardiomoycytes were divided into three groups:control group,model group and experimental group.The cardiomyocytes apoptosis rates were detected by Hoechest33342/PI and Annexin-FIFC/PI double staining.The rate of the cardiomyocytes survival rate was detected by MTT detection kit.The expression of B-type natriuretic peptide(BNP)and transforming growth factor-β1(TGF-β1)mRNA and protein of the cardiomyocytes were detected by qRT-PCR and Western blot respectively.Results Compared with the control group,the cardiomyocytes apoptosis rate in the model group was higher;Compared with the model group,the cardiomyocytes apoptosis rate in the experimental group was reduced(all P<0.05).Compared with the control group,the expression of BNP mRNA and protein in the model group were down-regulated,and the expression of TGF-β1 mRNA and protein were up-regulated(all P<0.05).Compared with the model group,the expression of BNP mRNA and protein in the experimental group were up-regulated,and the expression of TGF-β1 mRNA and protein were down-regulated(all P<0.05).Conclusion It indicated that EPCs-CM inhibited ADR-induced cardiomyocytes apoptosis possibly via the suppression of the expression of TGF-β1 and the promotion of the expression of BNP.