栀子苷对缺氧/复氧小胶质细胞TLR4通路的影响  

Geniposide on TLR4 pathway in cultured rat microglia with hypoxia/reoxygenation

作　　者：侯金才[1] 张鹏[1] 刘建勋[1] 韩笑[1] 李丹[1] 王秀辉[1] 

机构地区：[1]中国中医科学院西苑医院实验中心,北京100091

出　　处：《中国药理学通报》2011年第6期769-773,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30772755;81073060);国家科技重大专项课题(No2009ZX09103-317);博士后科学基金资助项目(No20100470514)

摘　　要：目的研究缺氧/复氧对原代培养的小胶质细胞TLR4受体及其通路中MyD88、NF-κBp65、p-ERK1/2、p-IκBα和p38的影响和不同浓度栀子苷的干预作用,以揭示栀子苷治疗脑缺血的分子生物学机制。方法原代培养小胶质细胞,采用缺氧/复氧的方式制备模型,用栀子苷125、250和500μmol.L-1 3个浓度梯度进行干预,用逆转录聚合酶链反应检测小胶质细胞TLR4 mRNA的表达,Western blot检测TLR4、p-IκBα、p38、p-ERK1/2蛋白,免疫荧光双染观察MyD88和NF-κB;结果缺氧/复氧激活了TLR4通路,并通过MyD88依赖途径激活了下游蛋白MyD88、NF-κBp65、p-ERK1/2、p-IκBα和p38;栀子苷250和500μmol.L-1组抑制了通路蛋白的活性;结论缺氧/复氧使TLR4通路蛋白磷酸化激活。栀子苷通过对TLR4通路蛋白的抑制发挥抗炎效应,促进脑缺血的恢复。Aim To study the effect of hypoxia / reoxygenation on TLR4,MyD88,NF-κBp65,p-ERK1/2,p-IκBα and the role of geniposide in the primary cultured microglia.To explore the molecular mechanism of the treatment of cerebral ischemia.Methods The hypoxia/reoxygenation model was established and treated with geniposide of 125,250 and 500 μmol·L-1,the expression of TLR4 mRNA with the reverse transcription polymerase chain reaction was detected,and the protein of TLR4,p-IκB,p38,p-ERK1/2 with Western blot,MyD88 and NF-κBp65 were observed with immunofluorescence double staining.Results The hypoxia/reoxygenation activates the TLR4 pathway and the protein of TLR4,MyD88,NF-κBp65,p-ERK1/2,p38 and p-IκB through the MyD88-dependent signaling,the group geniposide of 250 and 500 μmol·L-1 inhibited the activation.Conclusions The hypoxia/reoxygenation activates the protein of TLR4 pathway.Geniposide plays anti-inflammatory role through inhibition of protein on the TLR4 pathway,and promotes the recovery of cerebral congestion.

关 键 词：缺氧/复氧 小胶质细胞 TLR4通路 栀子苷 MyD88 NF-κBp65 P-ERK1/2 p-IκBα p38 

分 类 号：R-332[医药卫生] R284.1