ClC-3氯通道调节氧糖剥夺所诱导的小胶质细胞的表型转化
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家自然科学基金青年基金(81402906);江苏省自然科学基金(BK20151566);江苏省高校自然面上项目(14KJB310010);江苏省大学生创新训练计划省级指导项目(201-84)


ClC-3 chloride channel regulated microglial phenotypic transformation induced by oxygen-glucose deprivation
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    目的:研究ClC-3氯通道在氧糖剥夺所致小胶质细胞表型转化中的作用。方法:应用小胶质细胞株(BV-2)制备氧糖剥夺模型,分别给予氯通道阻断剂DIDS和NPPB预处理BV-2细胞。通过MTT活性测定确定BV-2细胞损伤及药物的有效浓度;通过实时荧光定量PCR法测定细胞表型相关分子,如M1型相关分子[肿瘤坏死因子--琢(tumor necrosis factor--琢,TNF--琢)-白介素-1β(interleukin 1-茁,IL-1-茁)-CD86]和M2型相关分子[转化生长因子-茁(transforming growth factor β,TGF-β)-CD206]mRNA水平的表达;通过RNA干扰的方法下调ClC-3的表达,再给予氯通道阻断剂DIDS和NPPB干预,进一步检测细胞的MTT活性,观察药物作用效果。结果:预先给予DIDS(1和10 μmol/L)和NPPB(1 μmol/L)能够改善氧糖剥夺所诱导的BV-2细胞MTT活性下降,抑制M1型相关分子如TNF--琢-IL-1-茁和CD86的表达并促进M2型相关分子(TGF--茁和CD206)的表达;通过RNA干扰下调ClC-3的表达,能够消除DIDS和NPPB的作用。结论:ClC-3氯通道参与调控氧糖剥夺所致小胶质细胞的表型转化,阻断ClC-3氯通道抑制氧糖剥夺诱导小胶质细胞向M1型转化。

    Abstract:

    Objective:To study the roles of ClC-3 chloride channel played in the microglial phenotypic transformation induced by oxygen-glucose deprivation. Methods:Microglia(BV-2 cell line) was applied to establish the oxygen-glucose deprivation (OGD) model,and then respectively pretreated with different concentrations of chloride channel blockers,including DIDS and NPPB. Cell damage and the effective concentration of drugs were determined by MTT activity. The mRNA level of cell phenotypic factors,such as M1 markers including tumor necrosis factor -琢(TNF--琢),interleukin 1-茁(IL-1-茁)and CD86,M2 markers containing transforming growth factor -茁(TGF--茁) and cell surface molecule CD206 were detected by real-time fluorescent quantitative PCR. Expression of ClC-3 was downregulated by small RNA interference,and then pretreated with chloride channels blockers-DIDS and NPPB. Finally,the effects of blockers were observed by the MTT activity. Results:Pretreatment with DIDS (1,10 μmol/L) and NPPB(1 μmol/L) could partially reverse the decrease of BV-2 cells viability induced by OGD. DIDS(1 μmol/L) and NPPB (1 μmol/L) pretreatment could reduce the mRNA level of TNF--琢,IL-1-茁 and CD86,they could also promote expression of TGF--茁 mRNA. However,the reversal effect of DIDS and NPPB could be abolished by the down-regulation of ClC-3 expression. Conclusion:ClC-3 chloride channel is essential for the phenotypic transformation of microglia caused by OGD. Blocking ClC-3 chloride channel could inhibit its transformation into M1 phenotype.

    参考文献
    相似文献
    引证文献
引用本文

董银凤,邢青青,常 瑶,秦 雪,张 华. ClC-3氯通道调节氧糖剥夺所诱导的小胶质细胞的表型转化[J].南京医科大学学报(自然科学版),2016,(10):1192-1197

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2016-05-26
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2016-10-17
  • 出版日期:
通知关闭
郑重声明