导痰洗心汤调控AMPK/SIRT1/NF-κB信号通路蛋白表达抑制阿尔茨海默病少突胶质细胞凋亡  

Daotan Xixin Decoction Inhibits Alzheimer's Disease Oligodendrocyte Apoptosis by Regulating AMPK/SIRT1/NF-κB Signaling Pathway Protein Expression

作　　者：汤子钦 卢建政[2] 文晓东[2] 周欣梅[2] 蒋媛静[2] 罗宁 TANG Zi-qin;LU Jian-zheng;WEN Xiao-dong;ZHOU Xin-mei;JIANG Yuan-jing;LUO Ning(Graduate School of Guangxi University of Chinese Medicine,Nanning530001;Neurology Department,Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning530011)

机构地区：[1]广西中医药大学研究生院,南宁530001 [2]广西中医药大学附属瑞康医院神经内科,南宁530011

出　　处：《中国中西医结合杂志》2024年第8期966-973,共8页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：广西自然科学基金项目(No.2021GXNSFAA196031)。

摘　　要：目的研究导痰洗心汤在阿尔茨海默病(AD)细胞模型中对MO3.13人少突胶质细胞凋亡的影响,并探索其调节机制.方法使用β淀粉样蛋白1-42(Aβ1-42)诱导MO3.13细胞构建AD细胞模型.将细胞分为空白对照组、AD细胞模型组、导痰洗心汤(低、中、高浓度)含药血清干预组及多奈哌齐干预组,每组细胞共3个样本.通过细胞增殖与活性检测试剂盒(CCK8)检测细胞活性,流式细胞仪检测细胞凋亡率,三磷酸腺苷(ATP)含量检测试剂盒检测细胞上清液中ATP含量,Western Blot检测磷酸化腺苷酸活化蛋白激酶(p-AMPK)、细胞沉默调节蛋白1(SIRT1)、B淋巴细胞瘤-2基因(Bcl-2)、核因子κB(NF-κB)及半胱氨酸天冬氨酸蛋白酶3(Caspase3)蛋白的表达丰度.结果与空白对照组比较,AD细胞模型组细胞活性降低(P<0.001),凋亡率增加(P<0.001),ATP含量减少(P<0.001),p-AMPK、SIRT 1和Bcl-2蛋白表达降低(P<0.001),NF-κB及Caspase3蛋白表达升高(P<0.001).与AD细胞模型组比较,高浓度导痰洗心汤组及多奈哌齐组细胞活性升高(P<0.01),凋亡率降低(P<0.001),ATP含量增高(P<0.001),p-AMPK、SIRT1及Bcl-2蛋白表达升高(P<0.001),NF-κB及Caspase3蛋白的表达降低(P<0.001).结论导痰洗心汤通过激活AMPK/SIRT1/NF-κB信号通路抑制AD少突胶质细胞的凋亡.Objective To investigate the effect of Daotan Xixin Decoction on MO3.13 human oligodendrocytic cell apoptosis in an Alzheimer's disease(AD)cell model,and to explore its regulatory mechanism.Methods In vitro model of Alzheimer's disease was established using MO3.13 cells induced by amyloid-β142(Aβ1-42).The cells were divided into the following groups:blank control group,AD cell model group,Daotan Xixin Decoction containing serum groups(low,medium and high concentration),and Donepezil containing serum group,with 3 samples in each group.Cell activity was detected using CCK8,apoptosis rate was detected by flow cytometry,and the content of adenosine triphosphate(ATP)in the cell supernatant was detected using an ATP content detection kit.Western Blot was used to detect the expression abundance of phosphorylated AMP-activated protein(p-AMPK),silent information regulator 1(SIRT1),B-cell lymphoma-2(Bcl-2),nuclear factor kappa B(NF-κB)and Caspase3 proteins.Results Compared with the blank control group,the cell activity in the AD cell model group decreased(P<0.001),the apoptosis rate increased(P<0.001),the ATP content decreased(P<0.001),the protein expression of p-AMPK,SIRT1,and Bcl-2 decreased(P<0.001),and the protein expression of NF-κB and Caspase3 increased(P<0.001).Compared with the AD cell model group,the cell activity of the high concentration Daotan Xixin Decoction group and Donepezil group improved(P<0.01),the apoptosis rate decreased(P<0.001),the ATP content increased(P<0.001),the protein expressions of p-AMPK,SIRT1,and Bcl-2 increased(P<0.001),and the protein expressions of NF-κB and Caspase3 decreased(P<0.001).Conclusion Daotan Xixin Decoction improves ATP content in AD oligodendrocytes by activating the AMPK/SIRT1/NF-κB signaling pathway and inhibits the inflammatory and pro-apoptotic effects of Aβ1-42 on AD oligodendrocytes.

关 键 词：导痰洗心汤 阿尔茨海默病 AMPK/SIRT1/NF-κB信号通路 少突胶质细胞 凋亡 中西医结合 

分 类 号：R285[医药卫生—中药学]