锌对辣椒素激活的神经元磷酸化胞外信号调节激酶表达的影响

The effect of Zinc on the expression of phosphorylated extracellular signal-regulate kinase in capsaicin stimulated neurons

目的研究锌对辣椒素激活的原代培养SD大鼠乳鼠大脑皮层磷酸化胞外信号调节激酶(phosphorylated extracellular signal-regulate kinase,pERK)表达的影响。方法原代培养大鼠乳鼠的皮层神经元,分为4组:对照组(Con,加入等量生理盐水),辣椒素组(N-CAP,加入等量生理盐水,24h后加入辣椒素200μmol·L-1),低锌组(L-CAP,加入氯碘羟喹5μmol·L-1,24h后加入辣椒素200μmol·L-1),高锌组(HCAP,加入氯化锌30μmol·L-1,24h后加入辣椒素200μmol·L-1)。应用免疫组织化学技术、免疫印迹技术和图像分析技术检测锌对辣椒素刺激的培养神经元中pERK表达的影响。结果辣椒素刺激后,pERK阳性神经元数量显著增多,平均光密度增大,蛋白表达量(免疫印迹)增多(P<0.01)。低锌组pERK蛋白表达量明显多于N-CAP组,而高锌组pERK蛋白表达量明显低于N-CAP组(P<0.01)。结论高锌抑制辣椒素激活的培养神经元pERK的表达。

Objective To study the effect of Zinc on the expression of phosphorylated extracellular signalregulate kinase（pERK） in capsaiciu sitmulated cultured neurons of new born SD rats. Methods The primary cultured cortical neurons of new born SD rats were divided into 4 groups： control group（Con, add saline only）, N-CAP group（add saline, then add capsaicin 200pmol · L^-1, 24 hours later）, L-CAP group（add clioquinol 51amol· L^-1, then add capsaicin 2001Jmol · L^-1, 24h later）, H-CAP group（add zinc chloride 30blmol · L^-1, then add capsaicin 200blmol· L^-1,24h later）. Immunohistochemistry, immunoblotting and image analysis were used to examin the expression of pERK in capsaicin sitmulated cultured neurons. Results The expression level of pERK in cultured neurons increased quickly after capsaicin treatment（P〈 0.01）, and increased greatly in L-CAP compared with N-CAP, but decreased greatly in H-CAP compared with N-CAP （P〈 0.01）. Conclusion High dose of Zinc inhibits the pERK expression in capsaicin stimulated neurons in vitro.