He Bin, Liu Shi-qing, Li Hao-huan
Department of Orthopedics, Renmin Hospital of Wuhan University, China.
Zhonghua Zheng Xing Wai Ke Za Zhi. 2010 Nov;26(6):444-7.
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.
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.
Morphological change was observed in PQQ treated Schwann cells. 1-500 nmol/L PQQ could up-regulate the expression of p-ERK1/2, and 1000 nmol/L had no effects, while 10 000 nmol/L exhibited inhibitory effect (P < 0.05). p-ERK1/2 increased to peak 1 h after PQQ added, and this up-regulation of p-ERK1/2 was inhibited by PD98059 (P < 0.05).
PQQ could affect morphology of Schwann cells and activation of ERK1/2. MEK inhibitor PD98059 could, block this activation. It suggests that MEK/ERK signal pathway should be involved in Schwann cells proliferation promoted by PQQ.
探讨丝裂原活化蛋白激酶(MEK)激酶级联反应、细胞外信号调节激酶(ERK1/2)信号通路在吡咯喹啉醌(PQQ)促进雪旺细胞增殖中的作用及其分子机制。
体外培养并纯化雪旺细胞,用S-100鉴定纯度。在培养基中加入不同时间和浓度的PQQ。采用蛋白质免疫印迹法检测ERK1/2及磷酸化ERK1/2的表达。用特异性抑制剂PD98059阻断MEK信号通路后,采用蛋白质免疫印迹法检测p-ERK1/2的表达。
观察到PQQ处理的雪旺细胞形态发生变化。1~500 nmol/L PQQ可上调p-ERK1/2的表达,1000 nmol/L无影响,而10 000 nmol/L表现出抑制作用(P<0.05)。加入PQQ后1 h,p-ERK1/2升高至峰值,PD98059可抑制p-ERK1/2的这种上调(P<0.05)。
PQQ可影响雪旺细胞形态及ERK1/2的激活。MEK抑制剂PD98059可阻断这种激活。提示MEK/ERK信号通路参与了PQQ促进雪旺细胞的增殖。