摘要
目的观察α-硫辛酸(alpha-lipoicacid,ALA)对6-羟基多巴胺(6-OHDA)诱导的PC12细胞帕金森病(PD)细胞模型的内质网应激及细胞凋亡相关因子表达的影响。方法使用传代的PC12细胞,CCK8法检测细胞活力,筛选6-OHDA和ALA的最佳给药浓度,将细胞随机分为对照组、6-OHDA(模型组)、6-OHDA+ALA组、ALA组,使用AnnexinⅤ-PE/7AAD试剂盒和流式细胞术检测各组细胞的凋亡率,提取药物干预后各组细胞总蛋白,Western blot法检测Bip/Grp78、CHOP、Active caspase3、Htr A2、α-syn、XIAP的蛋白表达。结果帕金森病细胞模型中6-OHDA的造模浓度为60μmol/L,ALA的最佳给药浓度为1μmol/L。各组细胞的凋亡率:对照组(3.50±0.97)%,6-OHDA组(24.66±3.54)%,ALA+6-OHDA组(15.59±2.97)%,ALA组(4.77±1.03)%,与对照组比较模型组的凋亡率明显升高(P<0.05),与模型组比较,ALA+6-OHDA组细胞凋亡率降低(P<0.05)。与对照组比较,6-OHDA组Bip/Grp78、CHOP、Active caspase3、Htr A2、α-syn的蛋白表达增高,XIAP的表达降低(P<0.05);与6-OHDA组比较,ALA+6-OHDA组Bip/Grp78、CHOP、Active caspase3、Htr A2、α-syn蛋白表达降低,XIAP的表达增高(P<0.05)。结论 ALA可提高PD细胞模型的细胞活力,减低细胞的凋亡率;ALA可能通过减少PD模型细胞α-syn集聚,抑制内质网应激启动的细胞凋亡途径,以及下调Omi/Htr A2抑制线粒体caspase凋亡途径,对多巴胺神经元起到一定的保护作用。
Objective To investigate the effects of alpha-lipoicacid（ALA） on endoplasmic reticulum stress（ERS） and apoptosis in acell model of Parkinson’s disease（PD）.Methods The cell vitality of PC12 cells was detected after treatment with different concentrations of 6-OHDA, and the best drug concentrations of 6-OHDA and ALA were screened with CCK8 kit. The PC12 cells were randomly divided into control group, 6-OHDA group（model group）, 6-OHDA＋ALA group, ALA group. The apoptosis rate of PC12 cells was detected by Annexin Ⅴ-PE/7AAD kit and flow cytometry（FCM）. The total protein was extracted after drug intervention to detect the expression of Bip/Grp78, CHOP, active caspase3, HtrA2, alpha-syn, XIAP by Western blot method.Results The best concentration of 6-OHDA was 60 μmol/L for establishing the model of Parkinson’s disease, and the optimal concentration of ALA was 1 μmol/L. FCM results showed that the apoptosis rate was（3.50±0.97）% in control group, （24.66±3.54）% in 6-OHDA group, （15.59±2.97）% in ALA＋6-OHDA group, and （4.77±1.03）% in ALA group. Compared with control group, the apoptosis rate was significantly increased in model group（ P 〈0.05）. Compared with model group, the apoptosis rate was significantly decreased in ALA＋6-OHDA group（ P 〈0.05）. Compared with control group, the protein expression levels of Bip/Grp78, CHOP, active caspase3, HtrA2, α-syn were significantly increased, and the expression of XIAP was decreased in 6-OHDA group（ P 〈0.05）.Compared with model group, the expression levels of Bip/Grp78, CHOP, active caspase3, HtrA2 and α-syn protein were decreased and the expression of XIAP was increased in ALA＋6-OHDA group（ P 〈0.05）.Conclusion ALA can improve cell vitality in PD model, and reduce the apoptosis rate. ALA can protect dopamine neurons by reducing the accumulation of α-syn in the cell model, inhibiting the cell apoptosis pathway initiated by endoplasmic reticulum stress, and down-regulateing the Omi/HtrA2 and inhibiti
作者
李艳霞
高华
王丹
李沛珊
王玉玲
杨新玲
LI Yanxia;GAO Hua;WANG Dan;LI Peishan;WANG Yuling;YANG Xinling(Department of Rehabilitation Medicine,Second Clinical Medical College,Xinjiang Medical University,Urumqi 830063,China;Department of Neurology,Fifth Clinical Medical College,Xinjiang Medical University;Department of Neurology,Second Clinical Medical College,Xinjiang Medical University;VIP Ward Of Internal Medicine,First Clinical Medical College,Xinjiang Medical University)
出处
《山西医科大学学报》
CAS
2018年第9期1045-1050,共6页
Journal of Shanxi Medical University
基金
国家自然科学基金-新疆联合基金项目(U1503222)
新疆维吾尔自治区科技支疆项目(201591160)
关键词
帕金森病
细胞模型
Α-硫辛酸
内质网应激
细胞凋亡
Parkinson＇s disease
cell model
alpha-lipoic acid（ALA）
endoplasmic reticulum stress （ERS）
apoptosis
