脑源性神经营养因子对H_2O_2诱导人动脉平滑肌细胞凋亡的作用  

Effects of Brain-derived Neurotrophic Factor on H_2O_2-induced Apoptosis of Human Arterial Smooth Muscle Cell

作　　者：陈建业[1] 刘戟[1] 王若菡[1] 刘智敏[1] 李忌[1] 陈俊杰[1] 

机构地区：[1]四川大学华西基础医学与法医学院生物化学与分子生物学研究所,成都610041

出　　处：《四川大学学报（医学版）》2003年第2期251-253,259,共4页Journal of Sichuan University(Medical Sciences)

摘　　要：目的　研究脑源性神经营养因子 (BDNF)对过氧化氢 (H2 O2 )诱导人动脉平滑肌细胞 (SMC)凋亡的作用。方法　 BDNF基因经脂质体介导转染 SMC, 型胶原基因调节元件 (COL IA1)驱动该基因在 SMC中异位表达 ,用 MTT法检测细胞增殖 ,吖啶橙 (AO)荧光染色观察细胞形态学变化 ,流式细胞仪检测细胞凋亡率 ,琼脂糖电泳检测 DNA图谱。结果　 10 0～ 80 0 μm ol/ L 浓度的 H2 O2 明显抑制 SMC增殖 ,2 0 0 μm ol/ L H2 O2 作用 2 4 h后 ,可见典型的细胞凋亡形态学及生物化学改变即胞浆固缩、核染色质断裂、DNA电泳呈梯形条带 ;经 BDNF基因修饰的 SMC则未显示上述改变。结论　 H2 O2 能有效诱导 SMC凋亡 ,转 BDNF基因的 SMC对 H2 O2Objective To investigate the protective effects of brain derived neurotrophic factor (BDNF) on hydrogen peroxide (H 2O 2) induced apoptosis of human arterial smooth muscle cells (SMC). Methods The hBDNF cDNA was transfected into the SMC mediated by Lipofect AMINE, and ectopic expression in SMC was driven by the regulatory elements of I type collagen gene(COLIA1). The cell proliferation was measured with a colorimetric assay based on the cleavage of the tetrazolium salt MTT. Microscopic analysis of cell apoptosis was performed with fluorescent stain of acridine orange (AO). Apoptosis rate of SMC was tested by flow cytometry(FCM) . DNA pattern was examined by agarose gel electrophoresis. Results Exposure of growing SMC to 100 800 μmol/L H 2O 2 for 24 h remarkably suppressed the cell proliferation. After treatment of the cells with 200μmol/L H 2O 2 for 24h, morphological and biochemical changes of classic apoptosis such as condensation of cytoplasm?fragmentation of the nulear chromatin, DNA 'ladder' pattern after electrophoresis were observed. Yet, the above mentioned apoptosis changes were not observed and the apoptosis rate was reduced obviously in the BDNF transfected SMC treated with the same concentration of H 2O 2. Conclusion H 2O 2 can induce apoptosis of SMC, but BDNF inhibits H 2O 2 induced SMC apoptosis markedly and has strong anti apoptotic activity.

关 键 词：脑源性神经营养因子 过氧化氢 动脉平滑肌细胞 细胞凋亡 

分 类 号：R363[医药卫生—病理学]