摘要
目的:探讨大黄素对缺糖/缺氧小胶质细胞的保护作用及机制。方法:建立缺糖/缺氧小胶质细胞模型;给予大黄素(20、40和80μmol/L)和/或过表达TLR4质粒(pcDNA-TLR4)对细胞进行干预;MTT法检测细胞活力;ELISA实验检测细胞上清液中乳酸脱氢酶(LDH)和肿瘤坏死因子α(TNF-α)的含量;流式细胞术检测细胞凋亡;Western blot检测细胞中Bax、Bcl-2、TLR4、p-IκB和IκB的蛋白水平。结果:与DMSO处理的缺糖/缺氧组相比,大黄素处理的缺糖/缺氧小胶质细胞的活力显著升高,LDH和TNF-α的含量、细胞凋亡率及Bax、TLR4、p-IκB的蛋白水平均显著降低,Bcl-2的蛋白水平显著升高(P<0.05);过表达TLR4可抑制大黄素处理的缺糖/缺氧小胶质细胞的细胞活力并增加其凋亡率。结论:大黄素对缺糖/缺氧诱导的小胶质细胞具有保护作用,其机制可能与TLR4/NF-κB信号通路失活有关。
AIM:To investigate the mechanism of emodin on the protection of glucose-deficient/anoxic microglia.METHODS:A microglia BV2 cell model induced by hypoglycemia/hypoxia(HH)was established.The glucose-deficient/anoxic cells treated with emodin were labeled as HH+emodin(20,40 and 80μmol/L)groups.The BV2 cells with TLR4 over-expression treated with emodin under hypoglycemia/hypoxia condition was labeled as HH+pcDNA-TLR4+emodin(40μmol/L)group.The cell viability was measured by MTT assay.Lactate dehydrogenase(LDH)and tumor necrosis factor-α(TNF-α)levels were detected by ELISA.The apoptosis was analyzed by flow cytometry.The protein levels of Bax,Bcl-2,TLR4,p-IκB and IκB were determined by Western blot.RESULTS:Compared with HH+DMSO group,the viability was significantly increased,the levels of LDH and TNF-αand apoptotic rate were significantly decreased,the protein levels of Bax,TLR4 and p-IκB were significantly decreased,the protein level of Bcl-2 was significantly increased in HH+emodin groups(P<0.05).Over-expression of TLR4 reversed the effect of emodin on promoting the viability and inhibiting apoptosis in the BV2 cells.CONCLUSION:Emodin has a protective effect on hypoglycemia/hypoxia induced microglia,and its mechanism may be related to the inactivation of TLR4/NF-κB signaling pathway.
作者
杨春澜
孟祥武
郑龙
陈娟
罗超
YANG Chun-lan;MENG Xiang-wu;ZHENG Long;CHEN Juan;LUO Chao(Department of Neurology,Minda Hospital Affiliated to Hubei University for Nationalities,Enshi 445000,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2019年第12期2285-2289,共5页
Chinese Journal of Pathophysiology
基金
湖北省自然科学基金资助项目(No.2015CFB476)
