上海交通大学学报（医学版）

›› 2012, Vol. 32 ›› Issue (10): 1296-.doi: 10.3969/j.issn.1674-8115.2012.10.004

• 论著（基础研究） • 上一篇    下一篇

梓醇活性成分对MPP+诱导的BV2细胞损伤的神经保护作用

王子玫, 许 刚, 张永芳, 胡雅儿   

  1. 上海交通大学 医学院细胞调控研究室, 上海 200025
  • 出版日期:2012-10-28 发布日期:2012-11-05
  • 通讯作者: 胡雅儿, 电子信箱: yaerhu@shsmu.edu.cn。
  • 作者简介:王子玫(1969—), 女, 助理研究员, 博士；电子信箱: cake_wangin21@yahoo.com.cn。
  • 基金资助:

    上海市教委科研创新项目（09YZ97）和上海市教委第五期重点学科资助项目（J50201）

Neuroprotective effects of Catalpol on BV2 cells injured by MPP+

WANG Zi-mei, XU Gang, ZHANG Yong-fang, HU Ya-er   

  1. Research Laboratory of Cell Regulation, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China
  • Online:2012-10-28 Published:2012-11-05
  • Supported by:

    Innovation Program of Shanghai Municipal Education Committee, 09YZ97；Leading Academic Discipline Project of Shanghai Municipal Education Committee, J50201

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摘要：

目的 观察大脑多巴胺神经营养因子（CDNF）在梓醇保护多巴胺胶质细胞中的作用。方法 ①将小鼠小胶质BV2细胞随机分为对照组、梓醇组、1-甲基-4-苯基吡啶离子（MPP+）模型组和MPP+模型+梓醇组，采用RT-PCR技术检测处理0、6、24、48和72 h后BV2细胞CDNF mRNA表达的变化。②采用终浓度分别为0.1、1和10  μmol/L的梓醇处理BV2细胞，24 h后加入MPP+，48 h后Western blotting检测CDNF蛋白表达；以正常细胞为对照组，设未加入梓醇预处理的MPP+模型组。③设对照组、MPP+模型组、梓醇组、抗CDNF抗体组和梓醇+抗CDNF抗体组；除对照组外，其余各组均经MPP+处理；采用[3H]-多巴胺放射分析法检测CDNF抗体阻断后梓醇对多巴胺摄取的影响。结果 ①在MPP+加入后的0～72 h，梓醇组和MPP+模型组的CDNF mRNA表达与对照组比较均无显著变化；与MPP+模型组比较，在MPP+加入后48 h，MPP+模型+梓醇组CDNF mRNA表达显著升高（P<0.01）。②MPP+加入后48 h，MPP+模型组CDNF蛋白表达量（0.679±0.013）低于对照组（1.009±0.015），差异有统计学意义(P<0.001)；以10 μmol/L梓醇预处理的BV2细胞CDNF蛋白表达量（0.812±0.011）显著高于MPP+模型组 (P<0.01)；0.1、1 μmol/L梓醇预处理的BV2细胞CDNF蛋白表达量与MPP+模型组比较无明显变化。③MPP+模型组多巴胺摄取力（63.5±2.5）低于对照组（99.9±0.8），差异有统计学意义（P<0.01）；梓醇组多巴胺摄取力（87.2±2.4）显著高于MPP+模型组（P<0.01）；梓醇+抗CDNF抗体组多巴胺摄取力（73.6±2.7）显著低于梓醇组（P<0.01），而高于抗CDNF抗体组（P<0.05）。结论 梓醇对MPP+诱导的BV2细胞损伤的神经保护作用与上调CDNF基因和蛋白表达有关。

关键词: 梓醇, 帕金森病, 大脑多巴胺神经营养因子, 多巴胺

Abstract:

Objective To explore the role of cerebral dopamine neurotrophic factor (CDNF) in the protection effect of Catalpol on the injured dopaminergic glial cells. Methods ①Mouse glial BV2 cells were randomly divided into control group, Catalpol group, 1-methyl-4-phenylpyridinium (MPP+) model group and MPP+ model+Catalpol group. The expression of CDNF mRNA in BV2 cells 0, 6, 24 and 48 h after treatment was detected by RT-PCR. ② BV2 cells were treated with 0.1, 1 and 10 μmol/L Catalpol respectively, MPP+ was added 24 h later, and the expression of CDNF protein was determined by Western blotting 48 h later. Normal cells were served as control group, and those without pretreatment with Catalpol were established as MPP+ model group. ③Control group, MPP+ model group, Catalpol group, anti-CDNF antibody group and Catalpol+ anti-CDNF antibody group were established, all groups were treated with MPP+ except control group, and the effect of Catalpol on dopamine uptake after CDNF antibody blockade was examined by [3H]-dopamine radiometry. Results ①Compared with control group, the expression of CDNF mRNA in Catalpol group and MPP+ model group was not significantly changed 0 to 72 h after administration of MPP+. However, the expression of CDNF mRNA in MPP+ model+ Catalpol group was significantly higher than that in MPP+ model group 48 h after administration of MPP+ （P<0.01）. ②The expression of CDNF protein in MPP+ model group was significantly lower than that in control group 48 h after administration of MPP+ （0.679±0.013 vs 1.009±0.015, P<0.001). The expression of CDNF protein in BV2 cells after pretreatment with 10 μmol/L Catalpol (0.812±0.011) was significantly higher than that in MPP+ model group （P<0.01）, while the expression of CDNF protein in BV2 cells after pretreatment with 0.1 and 1 μmol/L Catalpol was not significantly changed compared with MPP+ model group. ③The dopamine uptake in MPP+ model group was significantly lower than that in control group （63.5±2.5 vs 99.9±0.8, P<0.01）. The dopamine uptake in Catalpol group (87.2±2.4) was significantly higher than that in MPP+ model group （P<0.01）. The dopamine uptake in Catalpol+anti-CDNF antibody group (73.6±2.7) was significantly lower than that in Catalpol group （P<0.01）, while was higher than that in anti-CDNF antibody group （P<0.05）. Conclusion The neuroprotection effect of Catalpol against MPP+ toxicity may be associated with the upregulation of expression of CDNF mRNA and protein in BV2 cells.

Key words: Catalpol, Parkinson´s disease, cerebral dopamine neurotrophic factor, dopamine