绿茶提取物EGCG对乙醇诱导原代海马神经元损伤的保护作用及其机制
| 作者: |
1杨海玉,
2刘勇,
3胡建新,
2戴艳枝
1 江西省人民医院临床医学研究所，南昌 330006 2 江西省人民医院病理科，南昌 330006 3 江西省人民医院药剂科，南昌 330006 |
| 通讯: |
刘勇
Email: jxpathology@126.com |
| DOI: | 10.3978/j.issn.2095-6959.2018.04.002 |
| 基金: | 江西省卫生厅中医药科研基金重点项目(2013Z005)；江西省杰出青年人才资助计划(20162BCB23056)。 |
摘要
目的：探讨绿茶提取物表没食子儿茶素没食子酸酯(epigallocatechin-3-gallate，EGCG)对乙醇诱导原代海马神经元损伤的作用及机制。方法：以原代培养的海马神经元为实验对象，设对照组、乙醇组、EGCG+乙醇组和EGCG组，采用MTT比色法检测细胞存活率以及Annexin-V APC/7-AAD双染法观察细胞凋亡状况，采用荧光实时定量PCR检测凋亡因子BCL2，BAX表达并进行氧化应激检测。结果：EGCG(100 µmol/L)可明显提高海马神经元存活率及抑制乙醇诱导的细胞凋亡(P<0.001)；另外，EGCG可促进海马神经元凋亡因子BCL2表达增高、BAX表达降低(P<0.001)，同时使抗氧化酶T-SOD和GSH-Px的酶活力明显增高(P<0.001)。结论：EGCG对乙醇诱导的原代海马神经元损伤具有一定的保护作用，其机制与抑制海马神经元凋亡及抗氧化应激损伤有关。
关键词：
表没食子儿茶素没食子酸酯；乙醇性痴呆；海马神经元；凋亡；氧化应激
Protective effect of green tea compound epigallocatechin-3-gallate on alcohol-induced primary-cultured hippocampal neuron injury and its mechanism
CorrespondingAuthor: LIU Yong Email: jxpathology@126.com
DOI: 10.3978/j.issn.2095-6959.2018.04.002
Foundation: This work was supported by the Traditional Chinese Medicine Key Project Foundation of Jiangxi Provincial Health Department (2013Z005) and Jiangxi Provincial Outstanding Youth Talents Scheme (20162BCB23056), China.
Abstract
Objective: To observe the protective effect of green tea compound epigallocatechin-3-gallate (EGCG) on alcohol-induced primary-cultured hippocampal neuron injury and discuss its mechanism. Methods: Hippocampal neurons were primary-cultured for 7 days and identified with immunofluorescence staining, divided into 4 groups: control group, ethanol group, EGCG + ethanol group and EGCG group. The cell viability was analyzed with MTT assay and the staining with Annexin-V APC/7-AAD was used to observe cell apoptosis. The expression of apoptosis factors including BCL2 and BAX was detected by Real-time fluorescent quantitative PCR, the activity of antioxidant enzymes such as T-SOD and GSH-Px in hippocampal neurons was also studied. Results: For primary-cultured hippocampal neurons, the cell viability was significantly increased and alcohol-induced cell apoptosis was apparently inhibited by treatment of EGCG (100 µmol/L) (P<0.001). Moreover, the result showed that anti-apoptotic factor BCL2 was significantly up-expressed and pro-apoptotic factor BAX was down-expressed, the enzyme activity of T-SOD and GSH-Px in hippocampal neurons was also obviously increased after EGCG treatment. Conclusion: The alcohol-induced injury of hippocampal neuron could be prevented by EGCG treatment, which is closely associated with its inhibition effects on alcohol-induced cell apoptosis and oxidative stress.
Keywords：
epigallocatechin-3-gallate; alcohol-associated dementia; hippocampal neuron; apoptosis; oxidative stress