ERK在LPS诱导大鼠雪旺氏细胞TNF-α表达中的作用  

The role of ERK in LPS induced TNF-αexpression on Schwann cells

作　　者：邵晓轶[1] 秦永伟[1] 王海波[2] 高永静[3] 程纯[2] 沈爱国[1] 

机构地区：[1]江苏省南通大学神经再生重点实验室,226001 [2]南通大学医学院微生物和免疫学教研室 [3]南通大学航海医学研究所

出　　处：《中华微生物学和免疫学杂志》2007年第6期485-489,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金(No.30300099;30500153);江苏省自然科学基金(BK2003035);江苏省高校自然科学研究项目(KJB180109;05KJB180100);江苏省社会发展科技指导性计划项目(BS2004526)

摘　　要：目的研究细胞外信号调节激酶(ERK)对脂多糖(IPS)诱导的大鼠雪旺氏细胞肿瘤坏死因子-α(TNF-α)表达的作用。方法采用Western blot检测细胞中ERK激酶的活性;用ERK的特异性抑制剂(PD98059)预处理细胞后,ELISA检测细胞中TNF-α水平;RT-PCR检测细胞中TNF-αmRNA的表达;用免疫荧光双标法检测ERK的表达定位。结果LPS可显著激活雪旺氏细胞中的ERK信号通路,其激活高峰在LPS刺激后30～45 min,60 min后ERK磷酸化水平开始逐渐下降;用PD98059预处理细胞后,可显著抑制细胞TNF-α以及TNF-αmRNA的产生;LPS刺激细胞后引起ERK磷酸化,PD98059可抑制该过程。结论ERK信号通路的激活可能是LPS诱导TNF-α表达的前提。通过阻断细胞内信号转导通路来减少TNF-α及其他细胞因子的产生,为抑制周围神经损伤后的炎症以及免疫反应发生提供了一条新的思路。Objective TO explore the roleof the extracellular signal regulated kinases （ERK） in lipopolysaccharide （LPS） induced TNF-α expression on Schwann cell. Methods The protein level of phosphorylated- ERK was determined by Western blot analysis. Schwann cells were pretreated with the inhibitor of ERK （PD98059） before incubation with LPS, intracellular concentration of TNF-α was detected by sandwich enzymehnked immunosorbent assay （ ELISA）, TNF-α mRNA was detected by RT-PCR. Intracellular location of phosphorylated-ERK was observed by immunofluorescent staining. Results LPS significantly activated ERK signal pathway. The peak of phosphorylated-ERK was detected after stimulation with LPS for 30-45 min. The level of phosphorylated-ERK began to fall-off after 60 min. Inhibition of ERK activation by using ERK inhibitor PD98059 predominantly suppressed expression of TNF-α or TNF-α mRNA. LPS had the effect on promoting phosphorylation of ERK. The process was distinctly blocked by PD98059. Conclusion Activation of ERK-pathway might be a precondition for the inducement of TNF-a expression. Reducing biosynthesis of TNF-α and other cytokines by blocking the cell signal pathway, which supports a new strategy against inflammatory and immune reaction after injury of peripheral nerve.

关 键 词：细胞外信号调节激酶 脂多糖 雪旺氏细胞 肿瘤坏死因子-α(TMF-α) 

分 类 号：R363[医药卫生—病理学]