ZnCl_2阻断电压门控质子通道对缺氧后小胶质细胞产生活性氧及促炎因子的影响  

The Effect of Blocking Voltage Gated Proton Channel Hv1 by ZnCl_2 on Production of ROS and Infl ammatory Factors in Microglia

作　　者：余颖[1] 喻志源[1] 谢敏杰[1] 王伟[1] 骆翔[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院神经内科,武汉430030

出　　处：《中国细胞生物学学报》2016年第7期829-835,共7页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81471200;91332108)资助的课题~~

摘　　要：为明确电压门控质子通道(voltage-gated proton channel,Hv1)对小胶质细胞的影响,研究了Hv1在正常、激活及抑制后对小胶质细胞活性氧、促炎因子产生的影响及机制,该研究使用原代培养的小胶质细胞,利用免疫荧光染色明确Hv1表达。用Hv1非特异性抑制剂Zn Cl2阻断该通道,用DCFH-DA染色、Real-time PCR观察糖氧剥夺(oxygen glucose deprivation,OGD)处理后小胶质细胞产生活性氧(reactive oxygen species,ROS)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)水平的变化。采用Western blot检测Hv1和Nox2蛋白质水平。实验结果显示,小鼠脑部小胶质细胞可检测到Hv1通道的表达;单纯ODG组缺氧后小胶质细胞ROS、TNF-α、IL-1β的产生明显增加;而OGD加Zn Cl2干预组,小胶质细胞ROS、TNF-α、IL-1β的产生较单纯OGD组显著降低;Western blot显示,OGD导致小胶质细胞中Hv1和Nox2蛋白质水平显著增加。以上结果表明,小鼠脑部小胶质细胞存在Hv1通道蛋白质,Zn Cl2阻断该通道可降低OGD诱导的小胶质细胞ROS和TNF-α、IL-1β的产生,该抑制作用可能与其抑制NADPH氧化酶的作用相关。To identify the expression of voltage-gated proton channel （Hvl） in microglia, and the effect of blocking Hvl on production of ROS and inflammatory factors in microglia, we used immunofluorescent staining to identify the protein level of Hvl in microglia. DCFH-DA and Real-time PCR were used to detect the production of ROS and inflammatory factors in microglia after OGD and Hvl blocking. Western blot was used to analyse the protein levels of Hvl and Nox2. The results showed that Hvl was detected in the microglia of C57BL/6 mice. The production of ROS and inflammatory factors were increased after OGD and reduced when Hvl was blocked by ZnCl2. The protein levels of Hvl and Nox2 were upregulated after OGD. Taken together, our results suggested that the Hvl protein existed in microglia. Blocking Hvl could reduce the production of ROS and inflammatory factors in microglia after OGD. This effect might be due to the inhibition ofNADPH oxidase.

关 键 词：小胶质细胞 电压门控质子通道 活性氧 促炎因子 

分 类 号：R741[医药卫生—神经病学与精神病学]