摘要
目的:观察热休克蛋白B1(heatshockproteinB1,HSPB1)对H2O2诱导的大鼠心肌细胞损伤过程中线粒体膜电位(DY)的影响,从线粒体水平阐明HSPB1保护心肌细胞抗氧化损伤机制。方法:以本室建立的稳定转染人HSPB1的大鼠心肌细胞系H9c2(HSPB1H9c2)和空载体转染的H9c2(CON)为实验对象,300μmol/LH2O2刺激2、4和8h后,倒置光学显微镜观察细胞形态学变化,荧光探针JC鄄1测定线粒体膜电位。结果:HSPB1高表达显著抑制了H2O2诱导的典型凋亡形态学改变;HSPB1高表达使线粒体膜电位升高42%以上(P<0.001);HSPB1高表达显著抑制线粒体膜电位下降,H2O2刺激2、4、8h后,HSPB1H9c2和CON的线粒体膜电位分别降至刺激前的69.5%和66.5%(P<0.05),74.4%和67.0%(P<0.05),70.7%和58.1%(P<0.05)。结论:HSPB1有效抑制了氧化应激诱导的以凋亡为主的细胞损伤和线粒体膜电位丧失,提示线粒体膜电位的稳定可能参与了HSPB1抗心肌细胞的氧化损伤机制。
Objective: To determine the effect of HSPB1 on the loss of mitochondrial membrane potential(D?追) in H2O2 induced rat cardiomyocytes oxidative injury. Methods: Established HSPB1 stable transfection rat cardiomyocytes cell line H9c2(HSPB1 H9c2) and empty vector transfected H9c2(CON) were cultured in 24-well plates, and treated by 300 靘ol/L H2O2 for 2,4 and 8 h. Cell morphology was analyzed by microscope and mitochondrial membrane potential was detected by JC-1. Results:Cell morphology changed significantly in CON after stimulated by 300 靘ol/L H2O2, but not in HSPB1 H9c2. HSPB1 expression increased mitochondrial membrane potential markedly in H9c2 by more than 42%(P<0.001). HSPB1 inhibited the loss of mitochondrial membrane potential induced by H2O2. After treated with H2O2 for 2,4 and 8h, mitochondrial membrane potential decreased to 69.5%and 66.5%(P < 0.05),74.4% and 67.0%(P < 0.05),70.7% and 58.1%(P < 0.05) in HSPB1 H9c2 and CON, respectively. Conclusion: HSPB1 inhibited apoptosis and prevented the loss of mitochondrial membrane potential induced by oxidative stress in rat cardiomyocytes H9c2, which suggests that stabilization of mitochondrial membrane potential may be involved in the protection of HSPB1 against oxidative stress in H9c2.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2005年第2期80-83,112,共5页
Journal of Nanjing Medical University(Natural Sciences)
基金
江苏省卫生厅"135"重点医学工程基金资助项目(135-14)
江苏省教育厅自然科学基金资助项目(03KJD320139)
关键词
热休克蛋白B1
心肌细胞
线粒体膜电位
氧化损伤
细胞凋亡
heat shock protein B1
cardiomyocytes
mitochondrial membrane potential
oxidative stress
apoptosis
