健脑增智饮对AD细胞模型的神经保护作用及对细胞凋亡的影响  

Effect of Jian Nao Zeng Zhi Yin on neuroprotection and apoptosis in AD cell model

作　　者：杨玲 翁旭亮 曾婷 王成银 刘青 邱铃铃 雷源 YANG Ling;WENG Xu-liang;ZENG Ting(Department of Encephalopathy,Affiliated Hospital of Traditional Chinese Medicine,Guangzhou Medical University,Guangzhou 510000,China)

机构地区：[1]广州医科大学附属中医医院脑病科,510000

出　　处：《中国现代药物应用》2024年第2期175-180,共6页Chinese Journal of Modern Drug Application

基　　金：广州医科大学附属中医医院中医药科研项目——青苗计划(项目编号:2019006);广东省中医药局中医药科研项目(项目编号:20201291)。

摘　　要：目的探讨健脑增智饮对阿尔茨海默病(AD)细胞模型的神经保护作用及对细胞凋亡的影响。方法将PC12细胞用20μmol/L的β-淀粉样蛋白(Aβ)1-42处理24 h后分为六组:正常细胞对照组(NC组)、AD细胞模型组(AD组)、阳性药物对照AD组(AD+Donepezil组)、健脑增智饮(低浓度、中浓度、高浓度)治疗AD组(AD+5μg/ml健脑增智饮组、AD+10μg/ml健脑增智饮组、AD+20μg/ml健脑增智饮组)。通过CCK-8检测法检测细胞活性;通过western蛋白印迹法检测凋亡因子相关蛋白Bax、Bcl-2的表达水平;应用流式细胞仪测定细胞凋亡情况。结果通过CCK-8检测法测定各组细胞活性结果显示:与NC组比较,AD组细胞活性显著降低(P<0.05);与AD组比较,AD+Donepezil组AD+20μg/ml健脑增智饮组细胞活性显著增高(P<0.05)。通过western蛋白印迹法测定凋亡因子相关蛋白表达水平结果显示:与NC组比较,AD组Bax表达水平显著上调,Bcl-2水平显著下调,Bcl-2/Bax显著下调(P<0.05);与AD组比较,AD+Donepezil组和AD+10μg/ml健脑增智饮组、AD+20μg/ml健脑增智饮组Bax水平显著下调(P<0.05),AD+Donepezil组和AD+20μg/ml健脑增智饮组Bcl-2/Bax显著上调(P<0.05)。应用流式细胞仪测定细胞凋亡情况结果显示:NC组细胞凋亡率为(7.870±0.241)%,AD组为(36.530±1.429)%,AD+Donepezil组为(10.690±1.257)%,AD+5μg/ml健脑增智饮组为(26.930±1.790)%,AD+10μg/ml健脑增智饮组为(23.800±1.136)%,AD+20μg/ml健脑增智饮组为(10.410±1.903)%。与NC组比较,AD组细胞凋亡率显著升高(P<0.05);与AD组比较,AD+Donepezil组和AD+5μg/ml健脑增智饮组、AD+10μg/ml健脑增智饮组、AD+20μg/ml健脑增智饮组细胞凋亡显著降低(P<0.05)。结论健脑增智饮可通过调控Bcl-2/Bax蛋白的表达,有效减少AD细胞的凋亡,提高细胞的活性,起到对AD细胞的神经保护作用,且高浓度(20μg/ml)健脑增智饮治疗组疗效更优,为中医治疗认知障碍疾病的进一步探索提供理论及临床依据。Objective To investigate the neuroprotective effect of Jian Nao Zeng Zhi Yin on Alzheimer's disease(AD)cell model and its effect on apoptosis.Methods PC12 cells were treated with 20μmol/L amyloid-β(Aβ1)-42 for 24 h and divided into six groups:normal cell control group(NC group),AD cell model group(AD group),positive drug control AD group(AD+Donepezil group)and Jian Nao Zeng Zhi Yin(low concentration,medium concentration and high concentration)treatment AD group(AD+5μg/ml Jian Nao Zeng Zhi Yin group,AD+10μg/ml Jian Nao Zeng Zhi Yin group,AD+20μg/ml Jian Nao Zeng Zhi Yin group).Cell viability was detected by CCK-8 assay;the expression levels of apoptotic factor-related proteins Bax and Bcl-2 were detected by Western blotting;and apoptosis was determined by flow cytometry.Results The cellular activity was tested by CCK-8 assay,and the results showed that the cellular activity in AD group was significantly decreased compared with the NC group(P<0.05);compared with the AD group,the cellular activity in AD+Donepezil group,AD+20μg/ml Jian Nao Zeng Zhi Yin group was also significantly increased(P<0.05).The expression levels of apoptotic factor-related proteins were tested by western blotting,and the results showed that the expression level of Bax was significantly up-regulated,Bcl-2 was significantly down-regulated and Bcl-2/Bax was significantly downregulated in AD group compared with the NC group(P<0.05);compared with the AD group,Bax in AD+Donepezil group,AD+10μg/ml Jian Nao Zeng Zhi Yin group and AD+20μg/ml Jian Nao Zeng Zhi Yin group was significantly down-regulated,while Bcl-2/Bax in AD+Donepezil group and AD+20μg/ml Jian Nao Zeng Zhi Yin group was significantly up-regulated(P<0.05).The apoptosis was tested by flow cytometry,and the results showed that the apoptosis rate was(7.870±0.241)%in NC group,(36.530±1.429)%in AD group,(10.690±1.257)%in AD+Donepezil group,(26.930±1.790)%in AD+5μg/ml Jian Nao Zeng Zhi Yin group,(23.800±1.136)%in AD+10μg/ml Jian Nao Zeng Zhi Yin group,(10.410±1.903)%in AD+20�

关 键 词：健脑增智饮 阿尔茨海默病 神经保护 细胞凋亡 

分 类 号：R285[医药卫生—中药学]