吉林大学学报(医学版) ›› 2017, Vol. 43 ›› Issue (01): 26-31.doi: 10.13481/j.1671-587x.20170106

• 基础研究 • 上一篇    下一篇

鹿茸多肽对冈田酸致大鼠海马神经元损伤时Tau、Bcl-2和Caspase-3表达的影响

黄晓巍1, 王艳玲2, 李哲2, 陶贵斌1, 张天柱1, 李朝政1, 徐岩1, 刘玥欣1, 张丹丹1, 曲晓波1   

  1. 1. 长春中医药大学中医药与生物工程研究开发中心, 吉林 长春 130117;
    2. 吉林大学基础医学院病原免疫细胞遗传学实验中心, 吉林 长春 130021
  • 收稿日期:2016-09-17 出版日期:2017-01-28 发布日期:2017-02-08
  • 通讯作者: 曲晓波,教授,博士研究生导师(Tel:0431-86172508,E-mail:quxiaobo0504@hotmail.com) E-mail:quxiaobo0504@hotmail.com
  • 作者简介:黄晓巍(1972-),女,吉林省长春市人,主任药师,中药药理学硕士,主要从事心脑血管疾病的中药药理学方面的研究。
  • 基金资助:

    吉林省教育厅“十二五”科学技术研究项目资助课题(吉教科合字[2014]第72号)

Influence of pilose antler polypeptides on expressions of Tau, Bcl-2 and Caspase-3 in rat HT22 cells injured by okada acid

HUANG Xiaowei1, WANG Yanling2, LI Zhe2, TAO Guibin1, ZHANG Tianzhu1, LI Chaozheng1, XU Yan1, LIU Yuexin1, ZHANG Dandan1, QU Xiaobo1   

  1. 1. Research and Development Center of Traditional Chinese Medicine and Bioengineering, Changchun University of Traditional Chinese Medicine, Changchun 130117, China;
    2. Department of Experimental Cencer of Pathogenobiology, Immunology, Cytobiology and Genetics, School of Basic Medical Sciences, Jilin University, Changchun 130021, China
  • Received:2016-09-17 Online:2017-01-28 Published:2017-02-08

摘要:

目的:观察冈田酸诱导大鼠海马神经元(HT22细胞)损伤时神经元微管相关蛋白(Tau蛋白)和凋亡相关因子的变化,探讨鹿茸多肽对神经细胞损伤的保护作用,阐明其相关作用机制。方法:大鼠HT22细胞体外传代培养,冈田酸诱导HT22细胞损伤。HT22细胞分为正常对照组、DMSO对照组、冈田酸损伤模型组和不同浓度鹿茸多肽组(终浓度分别为50、500及1000 mg·L-1),37℃、5% CO2孵育24 h。采用免疫细胞化学染色法、Western blotting法和RT-PCR法检测各组细胞中Tau蛋白磷酸化水平及凋亡相关因子Bcl-2和Caspase-3的表达。结果:免疫细胞化学染色法染色,磷酸化Tau蛋白的表达呈棕黄色颗粒,与正常对照组比较,冈田酸损伤模型组细胞中棕黄色颗粒明显增多;与冈田酸损伤模型组比较,500和1000 mg·L-1鹿茸多肽组细胞中棕黄色颗粒明显减少。Western blotting法检测,与冈田酸损伤模型组比较,各浓度鹿茸多肽组细胞中Tau蛋白表达水平明显降低(P<0.05或P<0.01),1000 mg·L-1鹿茸多肽组细胞中Bcl-2表达水平明显升高(P<0.05),各浓度鹿茸多肽组细胞中Caspase-3表达水平均无明显变化(P>0.05)。RT-PCR法检测,与冈田酸损伤模型组比较,各浓度鹿茸多肽组细胞中Tau mRNA表达水平明显降低、Bcl-2mRNA表达水平明显升高(P<0.05或P<0.01),500和1000mg·L-1鹿茸多肽组细胞中Caspase-3mRNA表达水平明显降低(P<0.05或P<0.01)。结论:鹿茸多肽可能通过抑制Tau过度磷酸化及抑制细胞凋亡调控机制,对冈田酸诱导的神经细胞损伤起到保护作用。

关键词: 冈田酸, HT22细胞, 微管相关蛋白, 鹿茸多肽, Bcl-2, Caspase-3

Abstract:

Objective: To observe the changes of neuronal microtubule-associated protein (Tau) and apoptosis related factors in the rat HT22 cells injured by okada acid,and to investigate the protective effect of pilose antler polypeptides (PAP) on the nerve cell injury, and to explore its mechanism.Methods: Okada acid was used to induce the HT22 cell injury in the rats after HT22 cells were cultured in vitro.The HT22 cells were divided into normal control group,DMSO control group,okada acid injury model group and different concentrations of PAP groups (the final concentrations were separately 50,500,and 1 000 mg·L-1),which were incubated for 24 h at in the condition of 37℃ and 5% CO2.The levels of Tau phosphorylation and the expressions of apoptosis related factors Bcl-2 and Caspase-3 in the HT22 cels in various groups were detected by immunocytochemical staining, Western blotting and RT-PCR methods.Results: The results of immunocytochemical staining showed that phospho-Tau was colored brown-yellow.The brown-yellow particles in the cells in okada acid injury model group were significantly increased compared with normal control group.The brown-yellow particles in 500 and 1 000 mg·L-1 PAP groups were significantly decreased compared with okada acid injury model group.The results of Western blotting method showed that the Tau protein expression levels in different concentrations of PAP groups were significantly decreased compared with okada acid injury model (P<0.05 or P<0.01).The expression level of Bcl-2 in 1 000 mg·L-1 PAP group was significantly increased compared with okada acid injury model group (P<0.05).The expression levels of Caspase-3 in different concentrations of PAP groups had no significant changes compared with okada acid injury model group (P>0.05).The results of RT-PCR showed that the levels of Bcl-2 mRNA in different concentrations of PAP groups were significantly increased compared with okada acid injury model group (P<0.05 or P<0.01);the expression levels of Caspase-3 mRNA in 500 and 1 000 mg·L-1 PAP groups were significantly decreased(P<0.05 or P<0.01).Conclusion: PAP could protect the nerve cells injured by okada acid through inhibiting the excessive phosphorylation of Tau protein and apoptosis mechanism of cells.

Key words: pilose antler peptid, okada acid, HT22 cells, Caspase-3, microtubule-associated protein, Bcl-2

中图分类号: 

  • R285.6