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The Effect of Blocking Voltage Gated Proton Channel Hv1 by ZnCl2 on Production of ROS and Inflammatory Factors in Microglia
Yu Ying, Yu Zhiyuan, Xie Minjie, Wang Wei, Luo Xiang*
Department of Neurology, Tongji Hospital, Tongji Medical College,Huazhong University of Science and Technology, Wuhan 430030, China
Abstract: To identify the expression of voltage-gated proton channel (Hv1) in microglia， and the effect of blocking Hv1 on production of ROS and inflammatory factors in microglia， we used immunofluorescent staining to identify the protein level of Hv1 in microglia. DCFH-DA and Real-time PCR were used to detect the production of ROS and inflammatory factors in microglia after OGD and Hv1 blocking. Western blot was used to analyse the protein levels of Hv1 and Nox2. The results showed that Hv1 was detected in the microglia of C57BL/6 mice. The production of ROS and inflammatory factors were increased after OGD and reduced when Hv1 was blocked by ZnCl2. The protein levels of Hv1 and Nox2 were upregulated after OGD. Taken together， our results suggested that the Hv1 protein existed in microglia. Blocking Hv1 could reduce the production of ROS and inflammatory factors in microglia after OGD. This effect might be due to the inhibition of NADPH oxidase.
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