舒马普坦通过细胞外信号调节激酶1／2和c—Jun氨基末端激酶信号通路下调三叉神经节内降钙素基因相关肽的表达  

Sumatriptan down-regulates calcitonin gene-related peptide expression via extraceUular signal- regulated 1/2 and c-dun N.terminal kinase signaling transduction pathways in rat trigeminal ganglion after organ culture

作　　者：罗国刚[1] 袁博博[1] 樊文静[1] 袁兴运[1] 霍康[1] 吕社民[2] 曹永孝[3] 徐仓宝 

机构地区：[1]西安交通大学医学院第一附属医院神经内科,710061 [2]西安交通大学医学院第一附属医院生物化学与分子生物系中心实验室,710061 [3]西安交通大学医学院第一附属医院药理学系,710061 [4]Division of Experimental Vascular Research, Institutionof Medicine, Lund University, 22184 Sweden

出　　处：《中华神经科杂志》2012年第7期511-515,共5页Chinese Journal of Neurology

基　　金：国家自然科学基金资助项目（30570631）;西安交通大学国际交流与合作重点项目（0702-07）

摘　　要：目的探讨舒马普坦对大鼠三叉神经节（TG）离体培养后降钙素基因相关肽（CGRP）表达水平的影响。方法采用TG离体培养模型，按数字随机表法将54个TG随机分为新鲜组（6个）、对照组（6个）和实验组（7个亚组，每亚组6个，共42个）。实验组TG培养液中分别加入4种不同浓度舒马普坦，细胞外信号调节激酶1／2（ERK1／2）信号通路阻滞剂U0126和PD98059，c—Jun氨基末端激酶（JNK）信号通路阻滞剂SP600125，孵育24h后免疫组织化学染色检测CGRP免疫反应（CGRP—ir）阳性细胞表达，实时定量PCR检测CGRP—mRNA表达量，Western blot定量磷酸化ERK1／2（pERK1／2）和JNK（pJNK）蛋白水平。结果离体培养24h后，TG内CGRP—ir（＋）细胞表达明显增高，0．1和0．5mg／ml舒马普坦组CGRP-ir（＋）细胞百分比、阳性面积、累积吸光度、平均吸光度、CGRP—mRNA水平较对照组明显下降（tPcP=8．652、26．382，tarca=6．220、13．917，tId=5．606、15．904，tM=2．661、21．748，tmRNA=8．032、15．675，均P〈0．05）；而0．02和2．50mg／ml舒马普坦与对照组CGRP表达差异无统计学意义。Western blot结果显示：0．50mg／ml浓度舒马普坦显著降低TG内pERK1／2、pJNK水平，降低程度分别接近于10μmol/L的U0126、PD98059和SP600125。结论一定浓度舒马普坦通过细胞内ERK1／2、JNK信号通路下调大鼠TG离体培养后CGRP的过度表达。Objective To explore the effects of sumatriptan on the modulation of calcitonin generelated peptide（CGRP） expression and its involving intracellular signaling transduction mechanisms in rat trigeminal ganglion（TG） after organ culture. Methods Using organ culture in vitro model, 54 isolated TGs of SD rats were randomly divided into fresh group （ n = 6 ）, control group （ n = 6 ） and experimental group （n = 42, 6 TGs for each subgroup）. Experimental group included seven subgroups, which were respectively pretreated with four different concentrations of sumatriptan, specific inhibitors of extracellular signal- regulated kinases 1/2 （ ERK1/2 ） pathway （ U0126 and PD98059 ）, and the inhibitor of c-Jun N-terminal kinase （JNK） （SP600125）. After co-cultured with above intervention agents for 24 h, CGRP- immunoreactivity （CGRP-ir） positive neurons and CGRP-mltNA expression levels were quantified by immunohistochemistry stain and real-time polymerase chain reaction, respectively. Phosphorylated ERK1/2 （pERK1/2） and JNK （pJNK） proteins levels were determined by Western-blotting method. Results The CGRP-ir （ ＋ ） neurons expression levels were significantly increased after 24 h organ culture. However,0. 10 and 0. 50 mg/ml concentrations of sumatriptan remarkably decreased the CGRP-ir （ ＋ ） neurons expression levels. The positive cell percentage, positive optic area, integrated optical density, mean optical density and CGRP-mRNA expression level in TG were significantly reduced than control groups （tpcP = 8.652, 26.382; taroa = 6.220, 13.917; tIA = 5.606,15.904; tMIA = 2.661, 21.748; tmRNA --8.032, 15. 675. P 〈 0. 05 ）. The CGRP-mRNA expressions were significantly down-regulated after co-incubation with concentration of 0. 50 mg/ml sumatriptan for 24 h in TG of SD rat （ P 〈 0. 05 ）. The levels of pERK1/2 and pJNK protein kinase detected by Western-blotting were significantly reduced by 0. 50 mg/ml concentration of sumatriptan, the degrees of which were clos

关 键 词：舒马普坦 降钙素基因相关肽 三叉神经节 信号传导 

分 类 号：R619.6[医药卫生—外科学]