摘要
目的探讨小檗碱体外对三转基因阿尔茨海默症(AD)小鼠海马神经元内质网应激及凋亡的影响。方法取出生24 h内三转基因AD乳鼠,分离培养海马神经元。将海马神经元分为对照组、1μmol/L小檗碱组和10μmol/L小檗碱组,培养第7天,后两组分别用1、10μmol/L小檗碱干预,而对照组给予等体积药物溶剂二甲基亚砜。采用TUNEL染色法检测各组神经元细胞凋亡率,采用MTT法检测各组神经元细胞存活率,采用乳酸脱氢酶(LDH)细胞毒性试剂盒检测各组神经元培养上清液LDH释放率,采用Western blotting法检测各组内质网应激相关蛋白[葡萄糖相关蛋白78(GRP78)]以及凋亡相关蛋白(Caspase-3、Bcl-2、Bax)表达。结果与对照组比较,1μmol/L小檗碱组、10μmol/L小檗碱组神经元细胞凋亡率、LDH释放率、Caspase-3及Bax蛋白表达均下降,细胞存活率、GRP78及Bcl-2蛋白表达均增加(P均<0. 05);且10μmol/L小檗碱组较1μmol/L小檗碱组上述指标变化更明显(P均<0. 05)。结论小檗碱体外可抑制AD小鼠海马神经元内质网应激,减少神经元凋亡,其机制可能与上调GRP78蛋白、抗凋亡蛋白Bcl-2表达及抑制Caspase-3蛋白、促凋亡蛋白Bax表达有关。
Objective To investigate the effects of berberine on endoplasmic reticulum stress and neuronal apoptosis in mice with three transgenic Alzheimer's disease.Methods Hippocampal neurons were isolated and cultivated by using a three-transgenic AD mouse within 24 h of birth.The experimental cells were divided into 3 groups:the control group,1μmol/L berberine group,and 10μmol/L berberine group,which were treated with the same volume of the drug solvent dimethylsulfoxide(DMSO),1μmol/L berberine,and 10μmol/L berberine for 7 days.The apoptosis of neurons in each group was detected by TUNEL staining,and MTT kit was used to detect the rate of cell death.Lactate dehydrogenase(LDH)cytotoxicity kit was used to LDH release rate.Western blotting was used to detect the expression levels of endoplas-mic reticulum stress-related protein GRP78 and apoptosis-related protein Caspsae-3,and Bcl-2 and Bax.Results Com-pared with the control group,the apoptosis rate,LDH release rate,Caspase-3 and Bax protein expression of hippocampal neurons in the 1μmol/L berberine group and 10μmol/L berberine group decreased,but the cell survival rate,GRP78 and Bcl-2 protein expression increased(all P<0.05);and 10μmol/L berberine group was more significant than 1μmol/L berberine group(P<0.05).Conclusion Berberine inhibits endoplasmic reticulum stress in mouse hippocampal neurons and reduces neuronal apoptosis in vitro by up-regulating the GRP78 protein,anti-apoptotic protein Bcl-2 expression,and inhibiting Caspase-3 protein and proapoptotic protein Bax expression.
作者
龚琼
欧阳雅帆
黄艳露
黄敏
GONG Qiong;OUYANG Yafan;HUANG Yanlu;HUANG Min(The Seventh Hospital Affiliated to Sun Yatsen University,Shenzhen 518107,China)
出处
《山东医药》
CAS
2018年第36期54-57,共4页
Shandong Medical Journal
基金
广东省中医药局科研项目(20181064)
关键词
阿尔茨海默症
小檗碱
三转基因小鼠
内质网应激
细胞凋亡
berberine
Alzheimer's disease
three transgenic mice
endoplasmic reticulum stress
apoptosis
