MEK/ERK信号通路在吡咯喹啉醌促雪旺细胞增殖中的作用

The extracellular signal-regulated kinase was promoted by Pyrroloquinoline Quinine in cultured Schwann cells

目的 探讨MEK/ERK信号通路在吡咯喹啉醌促雪旺细胞增殖过程中的作用. 方法 体外培养雪旺细胞,S-100免疫荧光鉴定;Western blot检测MEK下游因子ERK1/2磷酸化激活形式（p-ERK1/2）的表达;MEK抑制剂（PD98059）阻断该通路后检测p-ERK1/2的表达;MTT法检测经PD98059阻断MEK通路后雪旺细胞的增殖情况. 结果 吡咯喹啉醌可激活雪旺细胞内MEK/ERK信号通路,在加入吡咯喹啉醌1 h后p-ERK1/2表达最高;吡咯喹啉醌在1～500 nmol/L范围内可使p-ERK1/2表达增加,1 000 nmol/L时与对照组比较差异无统计学意义,10 000 nmol/L时则表现为抑制作用（P＜0.05）;经PD98059阻断MEK通路后p-ERK1/2的上调效应消失（P＜0.05）.而且加入PD98059阻断MEK通路后吡咯喹啉醌对雪旺细胞的促增殖效果减弱. 结论 吡咯喹啉醌可激活雪旺细胞MEK/ERK信号通路,且该通路在吡咯喹啉醌促雪旺细胞增殖过程中发挥作用.

Objective To investigate the effect of mitogen-activated protein kinase（MEK）kinase cascade,extracellular signal-regulated kinase（ERK1/2）signal pathway on Schwann cells proliferation promoted by Pyrroloquinoline Quinine（PQQ）and its molecular mechanisms. Methods Schwann cells were cultured and purified in vitro.The purity was identified by S-100.Different time and concentration of PQQ was added into culture medium.The expression of ERK1/2 and phosphorylated-ERK1/2 was detected by western blot.The expression of p-ERK1/2 after blocking of MEK signal pathway by specific inhibitor PD98059 was detected by western blot. Results Morphological change was observed in PQQ treated Schwann cells.1 ～ 500 nmol/L PQQ could up-regulate the expression of p-ERK1/2,and 1 000 nmol/Lhad no effects,while 10 000 nmol/L exhibited inhibitory effect（P〈0.05）.p-ERKI/2 increased to peak 1 h after PQQ added,and this up-regulation of p-ERKI/2 was inhibited by PD98059（P〈0.05）. Conclusions PQQ could affect morphology of Schwann cells and activation of ERKI/2.MEK inhibitor PD98059 could block this activation.It suggests that MEK/ERK signal pathway should be involved in Schwann cells proliferation promoted by PQQ.