MRP8通过TLR4/NF-κB信号途径刺激培养星形胶质细胞分泌IL-1β

MRP8 stimulated IL-1β serection in cultured astrocyte via TLR4/NF-κB pathway

目的:探讨MRP8刺激星形胶质细胞(astrocyte,AS)释放IL-1β的信号途径。方法:利用MRP8刺激培养AS,运用Lipo2000脂质体转染发卡样小干扰RNA(shRNA)使TLR4沉默、二硫代氨基甲酸吡咯烷(PDTC)预处理阻断NF-κB,运用免疫印迹(Western Blot)、电泳迁移率变动分析(EMSA)、酶联免疫吸附试验(ELISA)、间接荧光免疫双标法等实验方法检测TLR4、NF-κB以及IL-1β的表达变化及相互作用关系。结果:利用MRP8刺激培养AS,细胞内的TLR4、NF-κB以及IL-1β表达增加,并且NF-κB由胞浆向胞核内转移,IL-1β在细胞上清液中含量增加;抑制TLR4可以下调MRP8所致的NF-κB和IL-1β表达增加,NF-κB向胞核内转移;抑制NF-κB只能下调MRP8所致的IL-1β表达增加。结论:MRP8可能通过TLR4/NF-κB信号途径促进刺激星形胶质细胞分泌IL-1β。

Objective：To study the pathway of IL-1β serection stimulated by MRP8 in astrocyte（AS）.Methods：MRP8were used to stimulate AS in vitro,lipofection transfectional shRNA were used to inhibit TLR4,PDTC were used to inhibit NF-κB.Western Blot,EMSA,ELISA and fluorescent double labelling were used to dectect the expressions of TLR4,NFkB and IL-1β,and analyze the relationship between these cytokines.Results：MRP8 increased TLR4,NF-κB and IL-1βexpressions in astrocyte,while NF-κB transferred into the nucleus,and IL-1β expression in cell supernatant also increased.TLR4 inhibition decreased the NF-κB and IL-1β expressions and NF-κB transferred into the nucleus induced by MRP8.NF-κB inhibition only decreased IL-1β expressions induced by MRP8.Conclusion：MRP8 stimulated IL-1βserection in cultured astrocyte via TLR4/NF-κB pathway.