肿瘤坏死因子α、白介素1β和舒马曲坦对大鼠三叉神经节离体培养后降钙素基因相关肽mRNA表达水平的影响  

Effects of TNF-α,IL-1β and sumatriptan on calcitonin gene-related peptide mRNA expression in rat trigeminal ganglion after organ culture

作　　者：罗国刚[1] 袁兴运[1] 袁博博[1] 雷莉[2] 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 [2]西安交通大学医学院遗传学与分子生物学系 [3]西安交通大学医学院药理学系 [4]Division of Experimental Vascular Research,Institution of Medicine,Lund University,Sweden 22184

出　　处：《西安交通大学学报（医学版）》2012年第3期276-280,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.30570631);西安交通大学国际交流与合作重点项目(No.0702-07)~~

摘　　要：目的探讨肿瘤坏死因子α(TNF-α)、白介素1β(IL-1β)和抗偏头痛药物舒马曲坦(Sumatriptan)对大鼠三叉神经节(TG)离体培养后降钙素基因相关肽(CGRP)mRNA表达水平的影响及其细胞内信号转导机制。方法将不同浓度的TNF-α、IL-1β、IL-2和Sumatriptan分别加入DMEM培养液中,与SD大鼠TG离体孵育24h;进一步将细胞内丝裂原激活的蛋白激酶(MAPK)信号转导系统ERK1/2、JNK、P38三条信号通路关键激酶特异性阻断剂PD98059、SP600125、SB239063加入DMEM培养液中,30min后分别加入有效浓度的炎性因子TNF-α和IL-1β后,与大鼠TG共同孵育24h,实时定量PCR(RT-PCR)法检测TG内CGRP-mRNA表达水平变化。结果 50μg/L的TNF-α、25μg/L的IL-1β可明显上调离体TG内CGRP-mRNA表达水平(P<0.05),0.1g/L和0.5g/L Sumatriptan能显著下调TG内CGRP-mRNA表达水平,而20～80μg/L的IL-2对CGRP-mRNA表达无影响。特异性阻滞剂SP600125、SB239063分别与50μg/L TNF-α、25μg/L IL-1β孵育24h后,CGRP-mRNA表达量较对照组显著降低(P<0.05)。结论 TG离体培养时,炎性因子TNF-α、IL-1β可显著上调CGRP-mRNA表达水平;一定浓度的舒马曲坦能下调CGRP-mRNA表达;JNK和P38信号转导通路参与了TNF-α、IL-1β诱导的TG内CGRP-mRNA上调过程。Objective To investigate the effects of TNF-α,IL-1β and sumatriptan on calcitonin gene-related peptide(CGRP) mRNA expression level in rat trigeminal ganglion(TG) and related mechanism of intracellular signaling transduction after organ culture.Methods CGRP-mRNA expression level was determined by quantified real-time polymerase chain reaction(RT-PCR) in TG after incubation with different concentrations of TNF-α,IL-1β,IL-2 and sumatriptan for 24 h.The specific extracellular signal-regulated kinase 1/2(ERK1/2) inhibitor PD98059,c-Jun N-terminal kinase(JNK) inhibitor SP600125 and protein kinase 38(P38) inhibitor SB239063 were respectively added into the medium 30 min prior to co-incubation with effective concentration of TNF-α and IL-1β in TG for 24 h;then the CGRP-mRNA expression level was compared among different groups.Results TNF-α of 50μg/L and IL-1β of 25μg/L significantly up-regulated CGRP-mRNA expression(P<0.05) whereas sumatriptan of 0.1g/L and 0.5g/L significantly down-regulated CGRP-mRNA expression in rat TG after 24-hour incubation.However,different concentrations of IL-2 did not show remarkable effects on CGRP-mRNA expression after 24-hour organ culture in TG.Both inhibitor SB239063 and SP600125 could remarkably decrease CGRP-mRNA over-expression induced by TNF-α of 50μg/L and IL-1β of 25μg/L,respectively(P<0.05). Conclusion The optimal concentration of TNF-α and IL-1β up-regulated whereas sumatriptan down-regulated CGRP-mRNA expression level in rat TG after 24 hours’ organ culture.JNK and P38 intracellular signaling transduction pathway may be involved in the CGRP-mRNA over-expression induced by inflammatory cytokines TNF-α and IL-1β.

关 键 词：降钙素基因相关肽 三叉神经节 肿瘤坏死因子Α 白介素1Β 舒马曲坦 

分 类 号：U[交通运输工程]