二苯乙烯苷对冈田酸致NG108-15细胞Tau蛋白磷酸化的影响
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篇名: 二苯乙烯苷对冈田酸致NG108-15细胞Tau蛋白磷酸化的影响
TITLE:
摘要: 目的:观察二苯乙烯苷(TSG)对冈田酸(OA)诱导致NG108-15细胞Tau蛋白磷酸化的影响,探讨该化合物抗阿尔茨海默病(AD)的可能机制。方法:以OA诱导NG108-15细胞复制AD细胞模型,采用MTT法检测经TSG低、中、高剂量(50、100、200 μmol/L)预处理后的细胞存活率,采用吖啶橙/溴乙锭双染色法检测细胞凋亡情况,采用Western blotting法和逆转录-聚合酶链反应法检测细胞周期蛋白依赖性激酶5(CDK5)、糖原合成酶激酶3β(GSK3β)蛋白及其mRNA以及Tau、磷酸化Tau(p-Tau)蛋白的表达情况,采用免疫荧光法检测CDK5、GSK3β、Tau蛋白的分布情况。结果:正常对照组细胞形态正常,未见或少见CDK5、GSK3β、Tau蛋白分布。模型组可见固缩或圆珠状的早期凋亡细胞,且CDK5、GSK3β、Tau蛋白分布明显增多,其细胞存活率显著降低,CDK5、GSK3β蛋白及其mRNA的相对表达量以及p-Tau与Tau相对表达量的比值(p-Tau/Tau)均显著升高(P<0.05或P<0.01)。经TSG预处理后,各给药组早期凋亡细胞和CDK5、GSK3β、Tau蛋白分布均有所减少,其细胞存活率均显著升高,中、高剂量组细胞CDK5蛋白、p-Tau/Tau以及各剂量组细胞CDK5 mRNA、GSK3β蛋白及其mRNA的相对表达量均显著降低(P<0.05)。结论:TSG对AD模型细胞具有一定的保护作用,这种作用与其提高细胞存活率,下调磷酸激酶CDK5、GSK3β的蛋白表达和基因转录水平,抑制Tau蛋白的磷酸化有关。
ABSTRACT: OBJECTIVE: To observe the effects of stilbene glycosidec (TSG) on okadaic acid (OA)-induced Tau protein phosphorylation in NG108-15 cells, and to investigate the potential anti-Alzheimer’s disease (AD) mechanism of this compound. METHODS: AD model of NG108-15 cells was induced by OA. The survival rate of NG108-15 cells was observed by MTT assay after pretreated with low-dose, medium-dose and high-dose of TSG (50, 100, 200 μmol/L). The apoptosis of NG108-15 cells was detected by AO/EB double fluorescence staining. The protein and mRNA expression of CDK5 and GSK3β, and the protein expression of Tau and p-Tau were detected by Western blotting assay and RT-PCR. The distribution of CDK5, GSK3β and Tau protein were detected by immunofluorescence. RESULTS: The normal morphology of NG108-15 cells was observed in normal control group, but CDK5, GSK3β and Tau protein were not found or few was found. Contracted or globular early apoptotic cells were observed in model gorup; the distribution of CDK5, GSK3β and Tau protein was increased, while survival rate of the cells was decreased; protein and mRNA expression of CDK5 and GSK3β as well as ratio of the relative expression of p-Tau to that of Tau (p-Tau/Tau) were all increased significantly (P<0.05 or P<0.01). After pretreatment of TSG, the distribution of early apoptotic cells as well as CDK5, GSK3β and Tau protein were all decreased to some extent in administration groups, while survival rates of the cells were increased significantly. Protein expression of CDK5 and p-Tau/Tau in medium-dose group and high-dose group as well as mRNA expression of CDK5, protein and mRNA expression of GSK3β in administration group were decreased significantly (P<0.05). CONCLUSIONS: TSG can protect against AD model cells, the effects of which may be associated with improving survival rate of the cells, down-regulating the protein expression and gene transcription level of phosphokinase CDK5 and GSK3β, inhibiting Tau protein phosphorylation.
期刊: 2019年第30卷第18期
作者: 谭俊杰,吴文雪,廖艳花,苏彦兆,李振中,黄健,黄忠仕
AUTHORS: TAN Junjie,WU Wenxue,LIAO Yanhua,SU Yanzhao,LI Zhenzhong,HUANG Jian,HUANG Zhongshi
关键字: 二苯乙烯苷;NG108-15细胞;阿尔茨海默病;Tau蛋白;磷酸化;周期蛋白依赖性激酶5;糖原合成酶激酶3β
KEYWORDS: Stilbene glycoside; NG108-15 cells; Alzheimer’s disease; Tau protein; Phosphorylation; CDK5; GSK3β
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