S100B在多柔比星诱导心肌细胞损伤中的表达及作用

Expression and role of S100B on the injury of cardiomyocytes induced by doxorubicin

目的:研究S100B在多柔比星诱导心肌细胞损伤中的表达变化及作用。方法:将体外培养的心肌细胞随机分为对照组,多柔比星损伤组(DOX组),多柔比星+阴性siRNA对照组(DNC组),DOX+S100B-siRNA组(DSB组)。用脂质体RNAi-MAX将S100B-siRNA转染到心肌细胞48 h后,多柔比星(2.0μmol/L)处理2 h。分别采用四甲基噻唑蓝(MTT法)、流式细胞术、蛋白质印迹法检测细胞存活率、凋亡率、S100B蛋白变化。结果:MTT测定显示,DNC组与DOX组存活率相比差异无统计学意义(P>0.05),DSB组与DOX组相比,其存活率增加(P<0.05)。流式细胞术分析表明,DOX组与DNC组凋亡率相比,差异无统计学意义(P>0.05);与DOX组相比,DSB组凋亡率降低(P<0.01)。蛋白质印迹检测S100B蛋白结果显示,DOX组与DNC组相比,差异无统计学意义(P>0.05);DSB组与DOX组相比,S100B蛋白表达下降(P<0.01)。结论:S100B在多柔比星诱导损伤的心肌细胞中表达明显增加并且可能促进心肌细胞凋亡。

Objective： To investigate the expression and the role of S100B on the injury of cardiomyocytes induced by doxorubicin.Methods： The cardiomyocytes from 1d SD rats were cultured in DMEMF-12 medium and then randomly divided into 4 groups： control group（CON）,doxorubicin damnified group（DOX）,doxorubicin＋negatived small interfering RNA control group（DNC）,and doxorubicin＋S100B small interfering RNA group（DSB）.S100B small interfering RNA was mediated by lipofectamineRNiMAX to transfect cardiomyocytes.The cardiomyocytes were treated with doxorubicin（2 μmol/L）for 2 h at 48 h post-transfection.The survival rate of cardiomyocytes was evaluated by MTT.The apoptosis rate was determined by flow cytometry（FCM）.The expression of S100B was detected by Western blot.Results： MTT assay showed that there was no significant difference in the survival rate of cardiomyocytes between DOX group and DNC group（P＆gt;0.05）.Compared with DOX group,the survival rate of cardiomyocytes in the DSB group was increased（P＆lt;0.05）;Flow cytometry showed that there was no significant difference in the apoptosis rate of cardiomyocytes between DOX group and DNC group（P＆gt;0.05）.Compared with DOX group,the apoptosis rate of cardiomyocytes in the DSB group was descended（P＆lt;0.01）;Western blot showed that there was no significant difference in the expression of S100B between DOX group and DNC group（P＆gt;0.05）,Compared with DOX group,the expression of S100B in the DSB group was descended（P＆lt;0.01）.Conclusion： The expression of S100B were significantly increased on the injury of cardiomyocytes induced by doxorubicin and might promote the apoptosis.