丝裂原活化蛋白激酶通路在GSK-3β对于巨噬细胞活化过程中的作用  

The role of mitogen-activated protein kinase pathway in GSK-3β for macrophage activation process

作　　者：张丹丹[1] 李珊珊[1] 李璐[1] 丁美[1] 段钟平[1] 陈煜[1] 

机构地区：[1]首都医科大学附属北京佑安医院人工肝中心,北京100069

出　　处：《胃肠病学和肝病学杂志》2015年第6期702-705,共4页Chinese Journal of Gastroenterology and Hepatology

基　　金：十二五重大专项课题资助项目(2012ZX10004904-003-00;2012ZX10002004-006;2013ZX10002002-006-001);中国肝炎基金会-天晴肝病研究基金课题资助项目(20120093);北京市医院管理局临床医学发展专项经费资助(XM201308)

摘　　要：目的探讨丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)通路在GSK-3β对于巨噬细胞活化过程中可能的作用。方法生长状态良好的RAW264.7细胞分为3组:对照组、脂多糖(LPS)组及GSK-3抑制剂SB216763干预组。在两个时间点(12 h、24 h),采用Western blotting法检测GSK-3β、p-GSK-3βser9、MAPK(c-jun氨基端激酶、细胞外信号调节激酶、p-38)表达情况,Elisa法检测细胞上清中TNF-α的变化,RT-PCR检测细胞中5-LO mRNA变化,电镜下观察RAW264.7细胞形态变化。结果 LPS组与对照组比较,p-GSK-3βser9/GSK-3β比值降低,活性升高;MAPK 3条信号通路:磷酸化的c-jun氨基端激酶(JNK)、细胞外信号调节激酶(ERK)、p38表达增多;TNF-α和5-LO mRNA表达增多(P<0.05);电镜下观察RAW264.7细胞出现形态改变,可见大量坏死物质。SB216763组与LPS组比较,p-GSK-3βser9/GSK-3β比值升高,活性降低;磷酸化JNK、ERK、p38表达出现不同时间不同程度改变;TNF-α和5-LO mRNA表达降低(P<0.05);电镜下观察RAW264.7细胞形态较规则,仅见少量坏死物质。结论 MAPK通路在GSK-3β对于RAW264.7细胞活化过程中发挥一定的作用。Objective To investigate the mitogen-activated protein kinase （MAPK） pathway whether plays a role in GSK-3β for macrophage activation process. Methods RAW264.7 cells grew well were divided into three groups： the experimental group, lipopolysaccharide （LPS） group and GSK-3 inhibitor SB216763 intervention group. At two points in time （12 hours, 24 hours） , the GSK-313, p-GSK-3βser9, MAPK （c-jun N-terminal kinase, extracellular signal-regu- lated kinase, p-38） expression were detected by Western blotting, The TNF-α in cell supernatants was detected by Elisa, changes of 5-LO mRNA in cells were detected by RT-PCR, RAW264.7 cell morphology was observed by electron microscopy. Results Compared with control group, p-GSK-3βser9/GSK-3β ratio decreased, activity increased, expres- sion of MAPK [ c-jun N-terminal kinase （JNK） , extracellular signal-regulated kinase （ERK） , p38 ] increased, expres- sion of TNF-α and 5-LO mRNA increased in LPS group （P 〈 0.05） , under electron microscopy, RAW264.7 ceils showed morphological changes, a large necrotic material. Compared with LPS group, p-GSK-3βser9/GSK-313 ratio in- creased, activity decreased, phosphorylation of JNK, ERK, p38 expression varied in different times, TNF-ct concentra- tion reduced in SB216763 group （P 〈 0.05）; under electron microscopy, RAW 264.7 cells showed morphological rules, only a small amount of necrotic material. Conclusion MAPK pathway plays a role in GSK-3β for RAW264.7 cells activation process.

关 键 词：糖原合成酶激酶-3Β RAW264.7细胞活化 丝裂原活化蛋白激酶 

分 类 号：R575[医药卫生—消化系统]