摘要
目的通过构建氯化钴(CoCl_(2))低氧模型,探究小胶质细胞在低氧条件下的活化状态,并探索其中机制。方法将小胶质细胞系BV2细胞,根据处理方式不同分为常氧组、CoCl_(2)4 h组(CoCl_(2)处理4 h)、CoCl_(2)6 h组(CoCl_(2)处理6 h)。采用DCFH-DA探针检测细胞中活性氧(ROS)的含量,运用Western Blot技术分析低氧诱导因子(HIF-1α)、TLR2/MyD88信号通路关键蛋白以及BV2细胞极化相关蛋白的表达变化。结果与常氧组相比,CoCl_(2)4 h组与CoCl_(2)6 h组活性氧水平明显升高,并且CoCl_(2)6 h组高于CoCl_(2)4 h组(P<0.05)。CoCl_(2)4 h组与CoCl_(2)6 h组的HIF-1α,TLR2/MyD88信号通路关键蛋白(TLR2、MyD88)表达量较常氧组明显升高(P<0.05)。CoCl_(2)4 h组的Arg1/iNOS比值与常氧组相比,轻微升高;而CoCl_(2)6 h组的Arg1/iNOS比值较常氧组明显降低(P<0.05)。结论在低氧条件下,细胞活性氧水平升高,通过TLR2-MyD88信号通路诱导炎症反应,BV2细胞被激活并在炎症损伤期间动态转化,早期以有益的M2型为主,而后期以有害的M1型为主,这初步揭示了低氧条件下小胶质细胞极化的相关机制。
Objective To explore the activation state of microglia cells under hypoxia and explore the mechanism through the construction of cobalt chloride(CoCl_(2))hypoxia model.Methods Microglia cell line BV2 cells were divided into the normal oxygen group,the CoCl_(2)4h group(CoCl_(2) treatment for 4 hours)and the CoCl_(2)6h group(CoCl_(2) treatment for 6 hours)according to different treatment methods.The content of reactive oxygen species(ROS)in cells was detected by DCFH-DA probe,and Western blot was used to analyze the expression changes of hypoxia-inducible factor(HIF-1α),the key proteins of TLR2/MyD88 signaling pathway and polarization related proteins in BV2 cells.Results Compared with that in the normal oxygen group,the level of ROS in the CoCl_(2)4h group and the CoCl_(2)6h group significantly increased,and that in the CoCl_(2)6h group was higher than that in the CoCl_(2)4h group(P<0.05).The expression levels of HIF-1α,TLR2/MyD88 signaling pathway key proteins(TLR2,MyD88)in CoCl_(2)4h and CoCl_(2)6h groups significantly increased compared with those in the normal oxygen group(P<0.05).The ratio of Arg1/iNOS in the CoCl_(2)4h group was slightly higher than that in the normal oxygen group,while the ratio of Arg1/iNOS in the CoCl_(2)6h group was significantly lower than that in the normal oxygen group(P<0.05).Conclusion Under hypoxic conditions,the level of cellular reactive oxygen species increases,which induces the inflammatory response through the TLR2-MyD88 signaling pathway.BV2 cells are activated and dynamically transformed during the inflammatory injury.The beneficial M2 type is dominant in the early stage,while the harmful M1 type is dominant in the late stage.Thus,the mechanism of microglia polarization under hypoxic conditions is preliminarily revealed.
作者
许娜
冯凌云
于燕
XU Na;FENG Lingyun;YU Yan(Binzhou Medical University,Yantai 264003,Shandong,P.R.China;Collaborative Innovation Center for Higher Education of Shandong Province,Yantai 264003,Shandong,P.R.China)
出处
《滨州医学院学报》
2021年第6期401-406,共6页
Journal of Binzhou Medical University
基金
山东省自然科学基金(ZR2021MH360,ZR2012HM001)。
