石榴补血糖浆对东莨菪碱诱导PC12细胞凋亡的保护作用及机制研究  

Protective effect of Pomegranate Blood-enriching syrup on apoptosis of PC12 cells induced by scopolamine and its mechanism

作　　者：赵丽蓉 黎晨 黄正元 尹海龙 成传芳 周本宏[1] ZHAO Lirong;LI Chen;HUANG Zhengyuan;YIN Hailong;CHENG Chuanfang;ZHOU Benhong(Department of Pharmacy,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院药学部,湖北武汉430060 [2]新疆维吾尔药业有限责任公司,新疆乌鲁木齐830026

出　　处：《陕西中医》2024年第5期588-592,共5页Shaanxi Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(31770381)。

摘　　要：目的:探讨石榴补血糖浆对东莨菪碱诱导大鼠肾上腺髓质嗜铬细胞瘤(PC12)细胞凋亡保护作用,探究石榴补血糖浆体外改善认知功能障碍的保护作用机制。方法:将不同浓度的石榴补血糖浆冻干粉细胞培养液作用于PC12细胞,采用CCK-8法检测细胞活性,根据细胞活性结果将后续实验分为空白对照组(NC)、模型组(SC)、石榴补血糖浆低、中、高剂量组(L、M、H:25、50、100μg/ml)、多奈哌齐对照组(D,10μmol/L);利用ELISA试剂盒检测细胞乙酰胆碱(Ach)、乙酰胆碱酯酶(AchE)、超氧化物歧化酶(SOD)和脂质过氧化产物丙二醛(MDA)的水平,流式凋亡术检测活性氧自由基(ROS)及细胞凋亡情况;Westernblot法检测凋亡相关蛋白Bcl-2、Bax、Caspase-3、Cleaved-Caspase-3等蛋白的表达情况。结果:石榴补血糖浆可显著增加PC12细胞的存活率,增加Ach、SOD的水平,降低ROS、MDA及AchE的水平,降低Bax/Bcl-2的比值和Cleaved-Caspase-3/Caspase-3蛋白的表达。结论:石榴补血糖浆可通过Bax/Bcl-2信号通路抑制细胞凋亡,减轻氧化应激,影响细胞内神经递质的水平发挥神经保护作用。Objective:To investigate the protective effect of Pomegranate Blood-enriching syrup on apoptosis of PC12 cells induced by scopolamine and the protective mechanism of Pomegranate Blood-enriching syrup on Alzheimer's disease(AD)in vitro.Methods:PC12 cells were cultured with different concentrations of Pomegranate Blood-enriching syrup freeze dried powders,and the cell activity was detected by CCK-8.According to the results of cell activity,the subsequent experiments were divided into blank control group,model group,low,middle and high dose groups(25,50,100μg/ml)and Donepezil control group(10μg/ml).The levels of acetylcholine(Ach),acetylcholinesterase(AchE),superoxide dismutase(SOD)and malondialdehyde(MDA)were detected by ELISA kit,reactive oxygen species(ROS)and apoptosis were detected by flow cytometry,and the expressions of apoptosis-related proteins such as Bcl-2,Bax,Caspase-3 and Cleaved-Caspase-3 were detected by Western blotting.Results:Pomegranate Blood-enriching syrup could significantly increase the survival rate of PC12 cells,increased the levels of Ach and SOD,decrease the levels of ROS,MDA and AchE,decreased the expression of Bax and Cleaved-Caspase-3 protein,and increased the expression of Bcl-2 protein.Conclusion:Pomegranate Blood-enriching syrup can inhibit apoptosis and reduce oxidative stress through Bax/Bcl-2 signal pathway,and can also play a neuroprotective effect by affecting the level of neurotransmitters.

关 键 词：石榴补血糖浆 神经保护 Bax/Bcl-2通路 认知障碍 细胞凋亡 PC12细胞 

分 类 号：R749.1[医药卫生—神经病学与精神病学]