补骨脂二氢黄酮甲醚对Aβ诱导PC12细胞损伤的保护作用及机制研究  

Protective Effect of Bavachinin A on PC12 Cells Injury Induced by Aβ and Its Mechanism

作　　者：杨柳 李爽 王业秋[1] 安丽凤[1] 李建民[1] 陈巧云[1] 黄敬文[1] 薛慧 YANG Liu;LI Shuang;WANG Yeqiu;AN Lifeng;LI Jianmin;CHEN Qiaoyun;HUANG Jingwen;XUE Hui(Jiamusi college,Heilongjiang University of Chinese Medicine,Jianiusi 154007 Heilongjiang,China)

机构地区：[1]黑龙江中医药大学佳木斯学院,黑龙江佳木斯154007

出　　处：《中药新药与临床药理》2021年第1期68-72,共5页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金面上项目(81274035);黑龙江中医药大学科研基金资助项目(201820);黑龙江省中医药管理局科研项目(ZHY16-102,ZHY18-049);黑龙江省卫生计生委科研项目(2017-578);黑龙江省大学生创新创业训练计划项目(201910228058)。

摘　　要：目的探讨补骨脂二氢黄酮甲醚对β淀粉样蛋白(β-amyloid protein,Aβ)诱导PC12细胞损伤的保护作用及其相关机制。方法采用Aβ(1.5×10-4mol·L-1)诱导的PC12细胞作为阿尔茨海默病(Alzheimer′s disease,AD)细胞模型,并以补骨脂二氢黄酮甲醚(1×10-6mol·L-1)、雌二醇(1×10-5mol·L-1)进行干预。实验设置空白组、Aβ组、补骨脂二氢黄酮甲醚组、雌二醇组。采用四甲基噻唑蓝(MTT)法检测细胞活力;流式细胞术检测细胞凋亡率;试剂盒检测细胞线粒体膜电位,以及超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)活性和丙二醛(MDA)含量;采用Western Blot法检测细胞中白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)蛋白表达水平。结果与空白组比较,Aβ组的细胞存活率、线粒体膜电位及SOD、GSH-PX活性明显降低(P<0.01),细胞凋亡率、MDA含量以及IL-1β、IL-6、TNF-a蛋白表达水平明显升高(P<0.01)。与Aβ组相比,补骨脂二氢黄酮甲醚组的细胞存活率、线粒体膜电位及SOD、GSH-PX活性明显升高(P<0.01),细胞凋亡率、MDA含量以及IL-1β、IL-6、TNF-α蛋白表达水平明显降低(P<0.01)。结论补骨脂二氢黄酮甲醚对Aβ诱导的PC12细胞损伤具有保护作用,可能与提高其抗氧化酶活性及线粒体膜电位,减少炎症因子分泌有关。Objective To investigate the protective effect of bavachinin A on PC12 cell injury induced by Aβ and its related mechanism.MethodsThe Alzheimer′s disease cell model was induced by applying Aβ(1.5×10-4 mol·L-1)in PC12 cells. After the preliminary experiment,Aβ(1.5×10-4 mol·L-1),bavachinin A(1×10-6 mol·L-1)and estradiol(1 × 10-5 mol · L-1) were selected for the subsequent experimental study. In this study, there were blank control group, Aβ group, bavachinin A group, estradiol group. Tetramethylthiazolium(MTT) was used to detect the viability of cells. Flow cytometry was used to detect apoptosis rate. Mitochondrial membrane potential detection kit(JC-1)was used to detect mitochondrial membrane potential. Superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and malondialdehyde(MDA)kits were used to detect the activities of antioxidant enzymes and content of MDA in cells. The expression levels of interleukin-1β(IL-1β),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were detected by Western Blot.ResultsCompared with the blank group,the cell survival rate,SOD,GSH-PX, mitochondrial membrane potential and activities in group Aβ were decreased(P<0.01), and the cell apoptosis rate, MDA content, expression levels of IL-1β, IL-6 and TNF-α protein were increased(P<0.01).Compared with group Aβ,the cell survival rate,mitochondrial membrane potentical,SOD and GSH-Px activities of bavachinin A group were increased(P<0.01),while the cell apoptosis rate,MDA content,IL-1β,IL-6 and TNF-α protein expression levels were decreased(P<0.01).ConclusionsThe protective effect of bavachinin A on PC12 cell damage induced by Aβ may be related to increasing the activity of antioxidases and mitochondrial membrane potential,reducing the secretion of inflammatory factors.

关 键 词：阿尔茨海默病 Β淀粉样蛋白 PC12细胞 补骨脂二氢黄酮甲醚 氧化应激 炎症因子 线粒体膜电位 

分 类 号：R285.5[医药卫生—中药学]