蛇毒神经生长因子对PC12细胞凋亡的保护作用  

Protection on the apoptosis of PC12 cells of NGF from Cobra venom

作　　者：何艳[1] 余文明[1] 陈崇宏[2] 

机构地区：[1]福建医科大学生化与分子生物学系,福州350004 [2]福建医科大学药学院,福州350004

出　　处：《海峡药学》2008年第10期36-39,F0004,共5页Strait Pharmaceutical Journal

基　　金：福建省教育厅基金资助项目(JA05252);福建医科大学教授基金资助项目(JS060026)

摘　　要：目的建立β-淀粉样蛋白诱导大鼠嗜铬瘤PC12细胞凋亡的体外AD细胞模型,研究蛇毒神经生长因子(NGF)的保护作用。方法以荧光显微镜观察细胞形态变化,四唑氮蓝(MTT)法,乳酸脱氢酶(LDH)释放法,流式细胞术等检测凝聚态β-淀粉样蛋白毒性片断Aβ25-35诱导的PC12凋亡,同时检测蛇毒NGF的干预作用。结果MTT法、LDH释放率显示凝聚态Aβ25-35对PC12细胞的毒性作用呈剂量依赖,20μmol·L-1Aβ25-35作用组细胞LDH释放率随NGF作用的浓度增高而降低;荧光显微镜观察20μmol·L-1Aβ25-35作用于PC12细胞出现凋亡改变,NGF预处理细胞的形态明显改善;流式细胞仪分析,Aβ25-35作用PC12细胞凋亡率具有剂量依赖关系,100ng.mL-1NGF预处理的细胞凋亡率降低(P<0.05)。结论凝聚态Aβ25-35对PC12细胞具有毒性作用;蛇毒NGF有对抗Aβ25-35致PC12细胞凋亡的作用。OBJECTIVE To set up AD cell model with amyloid β-peptide fragment 25-35（Aβ25-35） that induces apoptosis in the rat pheochromocytoma lines PC12 cells,and to explore the protective effect of NGF from cobra venom.METHODS The PC12 cells were cultured,treated with Aβ25-35 or/and NGF and collected for apoptotic analysis.The cytotoxicity of Aβ25-35 was assessed by the MTT and LDH release assay.Morphological change was screened by fluorescence microscopy and the apoptosis ratio analyzed by flow cytometry.RESULTS MTT and LDH release assay showed that the cytotoxity of Aβ25-35 was in a dose-and time-dependent manner to PC12 cells,and at the concentration of 20μmol·L^-1Aβ25-35,with the concentration of NGF increasing,cell viability was increasing.Screened by fluorescence microscopy it was observed that treatment with 20μmol·L^-1Aβ25-35 on PC12 cells for 48h induced cell morphological changes,and analyzed by flow cytometry,apoptosis ratio in PC12 cells evidenced increased in a dose-dependent manner,and when pretreated with 100ng·mL-1 NGF,the apoptocis ratio decreased（P〈0.05）.CONCLUSIONS Aβ25-35 efficiently induced apoptosis in PC12 cells.NGF from cobra venom had a protective effect against Aβ25-35-induced apoptosis in the PC12 cells.

关 键 词：蛇毒 神经生长因子 凋亡 

分 类 号：R97[医药卫生—药品]