Page 42 - 2019年9月第30卷第18期

二苯乙烯苷对冈田酸致NG108-15细胞Tau蛋白磷酸化的影响 Δ

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谭俊杰 ，吴文雪 ，廖艳花 ，苏彦兆 ，李振中 ，黄 健 ，黄忠仕 （1.广西中医药大学药学院，南宁 530200；2.
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右江民族医学院科技处，广西 百色 533000）
中图分类号 R285.5 文献标志码 A 文章编号 1001-0408（2019）18-2485-06
DOI 10.6039/j.issn.1001-0408.2019.18.07
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。
关键词 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

Effects of Stilbene Glycoside on Okadaic Acid-induced Tau Protein Phosphorylation in NG108-15 Cells
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TAN Junjie ，WU Wenxue ，LIAO Yanhua ，SU Yanzhao ，LI Zhenzhong ，HUANG Jian ，HUANG Zhongshi（1.
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College of Pharmacy，Guangxi University of TCM，Nanning 530200，China；2. Technology Department，
Youjiang Medical University for Nationalities，Guangxi Baise 533000，China）
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 GSK3 β ，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，GSK3 β 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.
KEYWORDS Stilbene glycoside；NG108-15 cells；Alzheimer’s disease；Tau protein；Phosphorylation；CDK5；GSK3β

Δ 基金项目：国家自然科学基金资助项目（No.81860709）；广西自 阿尔茨海默病（Alzheimer’s disease，AD）亦称老年
然科学基金资助项目（No.2016GXNSFAA380249） 性痴呆，是一种以近期记忆障碍为主要临床症状，β淀粉
＊硕士研究生。研究方向：抗老年痴呆药物研发。E-mail：
样蛋白沉积致老年斑、Tau 蛋白过度磷酸化致神经元纤
412544784@qq.com
维缠结以及神经元丢失伴胶质细胞增生为特征性病理
# 通信作者：教授，硕士生导师，博士。研究方向：抗老年痴呆药
[1]
物研发。电话：0776-2876020。E-mail：hzs1004@163.com 改变的一种进行性神经退行性疾病 。其中，Tau蛋白为

中国药房 2019年第30卷第18期 China Pharmacy 2019 Vol. 30 No. 18 ·2485 ·