上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (6): 769-.doi: 10.3969/j.issn.1674-8115.2011.06.019

• 论著（基础研究） • 上一篇    下一篇

中脑腹侧多巴胺能神经元对氧化应激反应的性别差异

陶青青1, 范晓兰2, 乐卫东1,2   

  1. 1.上海交通大学 医学院附属瑞金医院神经科, 上海 200025； 2.中国科学院上海生命科学研究院/上海交通大学医学院健康科学研究所, 上海 200025
  • 出版日期:2011-06-28 发布日期:2011-06-27
  • 作者简介:陶青青（1985—）, 男, 硕士生；电子信箱: jasontaoqing@126.com。

Gender differences of oxidative stress reactions in ventral mesencephalic dopamine neurons

TAO Qing-qing1, FAN Xiao-lan2, LE Wei-dong1,2   

  1. 1.Department of Neurology, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China；2.Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences &|Shanghai Jiaotong University School of Medicine, Shanghai 200025, China
  • Online:2011-06-28 Published:2011-06-27

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摘要：

目的 研究中脑腹侧（VM）多巴胺能神经元对氧化应激反应的性别差异。方法 原代培养不同性别Balb/c胎鼠的VM多巴胺能神经元，分为对照组（未处理）、鱼藤酮组（25 nmol/L鱼藤酮处理8 h）、雌激素+鱼藤酮组（用1×10-7 mol/L的17β-雌二醇预处理1 h后，加25nmol/L鱼藤酮处理8 h）。采用免疫组织化学法观察不同性别神经元的损伤程度；MTT法检测细胞存活率；Western blotting检测不同性别细胞酪氨酸羟化酶（TH）蛋白的表达。结果 免疫组织化学检测结果显示：鱼藤酮组雄性细胞较雌性细胞损伤大，TH阳性细胞更明显。MTT检测结果显示:鱼藤酮组雄性细胞存活率为66%，雌性细胞存活率为75%，差异有统计学意义（P＜0.05）；雌激素+鱼藤酮组雄性细胞存活率为81%，雌性细胞存活率为86%，高于相同性别的鱼藤酮组（P＜0.05）。Western blotting检测结果显示：鱼藤酮组雄性细胞的TH蛋白水平较雌性细胞明显下降（P＜0.05）；雌激素+鱼藤酮组TH蛋白水平高于相同性别的鱼藤酮组（P＜0.05）。结论 VM多巴胺能神经元对氧化应激反应存在性别差异，雄性细胞对鱼藤酮的刺激较雌性细胞更敏感，雌激素对两种性别的细胞均有保护作用。

关键词: 多巴胺能神经元, 氧化应激, 性别差异

Abstract:

Objective To investigate the gender differences of oxidative stress reactions in ventral mesencephalic (VM) dopamine neurons. Methods VM dopamine neurons of male and female Balb/c fetal mice were cultured, and were divided into control group (without treatment), rotenone group (treated with 25 nmol/L rotenone for 8 h) and estrogen+rotenone group (treated with 1×10-7 mol/L 17 β-estradiol for 1 h and 25 nmol/L rotenone for 8 h). Immunohistochemistry was employed to observe the neuron injury of mice of different genders, MTT assay was adopted to measure the cell viability, and Western blotting was used to detect the expression of tyrosine hydroxylase (TH) protein in cells of mice of different genders. Results Immunohistochemistry revealed that cell injury of male mice was more severe than that of female mice, and cells of male mice with positive TH were more significant than those of female mice with positive TH in rotenone group. MTT assay indicated that there were significant differences in cell viability between male mice (66%) and female mice (75%) in rotenone group (P＜0.05), and cell viability of male mice (81%) and female mice (86%) in estrogen+rotenone group was significantly higher than that of mice of corresponding genders in rotenone group (P＜0.05). Western blotting demonstrated that the expression of TH protein in cells of male mice was significantly lower than that in cells of female mice in rotenone group (P＜0.05), and the expression of TH protein in estrogen+rotenone group was significantly higher than that of corresponding genders in rotenone group (P＜0.05). Conclusion There exist gender differences of oxidative stress reactions in VM dopamine neurons. Male is more vulnerable than female in respond to rotenone, and estrogen has protective effects on both male and female neurons.

Key words: dopamine neuron, oxidative stress, gender differences