BDNF对H_2O_2诱导人成纤维细胞凋亡的实验研究  

The experimental study of Brain derived Neurotrophic Factor on H_2O_2-induced Apoptosis in Human fibroblast

作　　者：陈建业[1] 王晓明[2] 唐中[2] 陈瑾歆 刘戟[3] 王若菡[3] 刘智敏[3] 陈俊杰[3] 

机构地区：[1]川北医学院生化教研室,四川南充637000 [2]川北医学院附属医院,四川南充637000 [3]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,四川成都610041

出　　处：《川北医学院学报》2004年第2期2-5,共4页Journal of North Sichuan Medical College

摘　　要：目的研究脑源性神经营养因子 (BDNF)对H2 0 2 诱导人成纤维细胞 (hFB)凋亡的作用。方法　将含人Ⅰ型胶原a1链(COLIAl)基因启动子和增强子元件并在其 3′端接BDNFcDNA的微基因pSVCEPBFCAT ,经脂质体介导转染hFB并驱动BDNF异位表达 ,采用四唑盐 (MTT)显色法检测细胞增殖 ,吖啶橙荧光染色显微镜观察细胞形态的改变 ,琼脂糖凝胶电泳检测染色质DNA断裂。结果　经 2 0 0 μmol/LH2 O2 作用 2 4h后 ,pSVCEPBFCAT转染组和对照组包括未转染hFB、空载体pSVCEPCAT转染hFB的细胞生长抑制率分别为 35 %和≥ 84 % ,p <0 .0 0 1;对照组hFB普遍可见凋亡细胞特有的形态学及生物化学改变 ,如胞浆和核固缩、染色质断裂、DNA电泳呈阶梯状条带 ,而pSVCEPBFCAT转染hFB未发现上述凋亡特征。结论　BDNF促进hFB增殖和拮抗H2 O2 诱导凋亡 ,明显抑制H2 O2 对hFB的细胞毒性。Objective The aim of this experiment is to investigate the effect of BDNF on hydrogen peroxide(H 2O 2)-induced apoptosis in the human fibroblasts (hFB) Medhods The minigene pSVCEPBFCAT containing the promoter and enhancer elements of the human al(I) collagen gene(COLIAl) at its 3′ terminus followed by hBDNF gene cDNA was transfected into hFB mediated by lipofect-AMINE and drived BDNF ectopic expression in the hFB. The cell proliferation was measured by the MTT assay. The morphological changes of apoptotic cells were observed by microscopy with fluorescent stain of acridine orange. The DNA fragmemtation was examined by agarose gel electrophoresis. Results After exposure of growing cells to 200 mol/L H 2O 2 for 24 hours, the inhibition rate of cell growth was respectively 35% and ≥84% in the pSVCEPBFCAT-transfected hFB and controls of non-transfected hFB and pSVCEPCAT-transfected hFB, p<0.001.The morphological and biochemical changes of apoptotic cells such as shrinkage of cytoplasm and nucleus, fragmentation of the chromatin and ladder pattern of DNA were commonly observed in the cell population of controls, but these apoptotic features were not discovered in the pSVCEPBFCAT-transfected hFB. Conclusion: BDNF markedly inhibits H 2O 2 cytotoxicity on hFB by promoting hFB proliferation and antagonizing H2O2-induced apoptosis.

关 键 词：BDNF H2O2 诱导 成纤维细胞 细胞凋亡 实验研究 

分 类 号：R741.02[医药卫生—神经病学与精神病学]