中国药理学与毒理学杂志2013，Vol.27Issue(3)：379-384,6.DOI:10.3867/j.issn.1000-3002.2013.03.013
远志皂苷对β淀粉样蛋白片段1-40诱导PC12细胞凋亡的抑制作用
Protection of tenuigenin against apoptosis of PC12 cells induced by amyloid beta-protein fragment 1-40
摘要
Abstract
OBJECTIVE To investigate the protective mechanism of tenuigenin (TEN) on the apoptosis of PC12 cells induced by amyloid beta-protein 1-40(Aβ1-40) in vitro.METHODS Aggregated Aβ1-40 25 μmol·L-1 which induced the apoptosis of PC12 cells was used to establish Alzheimer's disease neuronal cell model.These model neurons were divided into Aβ1-40 model group and TEN 50,100 and 200 μmol·L-1 groups.At the same time,normal cell control group was established without Aβ1-40 pretreatment.The survival rate of PC12 cells was detected by MTT assay.The apoptosis rate of PC12 cells was detected by flow cytometery with Annexin-Ⅴ/PI double staining.The rates of positive cells expressed Bcl-2,Bax and cytochrome c (Cyt c) were observed by immunocytochemical method.The expression level of Cyt c was detected through Western blotting analysis.RESULTS Compared with normal control group,survival rate of PC12 cell decreased to (31 ± 7)％ (P ＜ 0.01),the positive rate of Bcl-2 was declined to (23.9 ± 1.9) ％ (P ＜ 0.01),the positive rate of Bax and Cyt c increased to (79.0 ±3.7) ％ and (49.2 ±3.6) ％ (P ＜ 0.01),respectively,and the ratio of Bcl-2/Bax was 0.30 in Aβ1-40 model group.Compared with model group,after 24 h incubation of with PC12 cells TEN 50,100 and 200 μmol·L-1,PC12 cell survival rate increased to (51 ±13)％,(64 ±7)％ and(84 ±10)％ (P＜0.01),respectively;the positive rate of Bcl-2 increased to (38.7 ±0.9)％,(53.7 ±1.6)％ and(60.3 ±0.8)％ (P ＜ 0.01),respectively; the positive rate of Bax was declined to (60.8 ± 1.9)％,(41.5 ±2.2)％ and (32.7 ±1.4)％(P＜0.01),and the ratio of Bcl-2/Bax increased to 0.64,1.29 and 1.84; the positive rate of Cyt c decreased to (45.4 ± 3.4) ％,(30.2 ± 2.2) ％ and (27.5 ± 1.0) ％ (P ＜0.05,P ＜0.01).Compared with normal control group,the apoptosis rate in PC12 cells and expression level of Cyt c protein were also obviously elevated (P＜0.01) ;and after 24 h incubation of TEN 50,100 and 200 μmol· L-1 with PC12 cells,the apoptosis rate in PC12 cells and expression level of Cyt c protein were lower than those in model group (P ＜ 0.01).CONCLUSION TEN has an obvious protecting effect against PC12 cell apoptosis induced by Aβ1-40.TEN may block the endogenous apoptosis pathway of PC12 cells by inhibiting the expression of Bax and Cyt c and increasing the expression of Bcl-2 and the ratio of Bcl-2/Bax.关键词
远志皂苷/β淀粉样蛋白片段1-40/PC12细胞/细胞凋亡/细胞色素cKey words
tenuigenin/ amyloid beta-protein fragment 1-40/ PC12 cells/ apoptosis/ cytochrome c分类
医药卫生引用本文复制引用
杨贤志,陈勤,陈庆林,金蓓蓓,叶海燕..远志皂苷对β淀粉样蛋白片段1-40诱导PC12细胞凋亡的抑制作用[J].中国药理学与毒理学杂志,2013,27(3):379-384,6.基金项目
The project supported by Natural Science Foundation of Education Bureau of Anhui Province (KJ2009A116) （KJ2009A116）
College Students of Anhui University Scientific Research Training Program (201135) （201135）
and Graduate Students' Academic Innovation Research Project of Anhui University (2010121) （2010121）
安徽省教育厅自然科学研究项目(KJ2009A116) （KJ2009A116）
安徽大学大学生科研训练计划项目(201135) （201135）
安徽大学研究生学术创新研究项目(2010121) （2010121）
