三叉神经节离体培养后降钙素基因相关肽调变的细胞内信号转导机制研究  

Intracellular Signaling Transduction Mechanism Study on the Modulation of Calcitonin Gene-related Peptide after Organ Culture of Rat Trigeminal Ganglion

作　　者：罗国刚[1] 雷莉[2] 樊文静[1] 吕社民[2] 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]遗传学与分子生物系,710061 [3]Division of Experimental Vascular Research,Institution of Medicine,Lund University

出　　处：《中国临床神经科学》2010年第2期113-118,共6页Chinese Journal of Clinical Neurosciences

基　　金：国家自然科学基金资助项目(编号:30570631);西安交通大学国际交流与合作重点项目(编号:062-10)

摘　　要：目的:探讨大鼠三叉神经节(TG)离体培养后降钙素基因相关肽(CGRP)表达增高的细胞内信号转导通路。方法:成年雄性SD大鼠TG在DMEM培养液离体培养12、24和48 h,应用免疫组化和实时定量PCR法分别检测CGRP阳性细胞数和mRNA表达水平变化。在培养液中分别加入一定浓度的炎症抑制剂地塞米松、转录水平特异性抑制剂放线菌素D、蛋白翻译水平特异性抑制剂环己酰胺,以及细胞内丝裂原激活的蛋白激酶(MAPK)信号转导系统ERK1/2、JNK、P38的3条信号通路上关键激酶的特异性阻断剂U0126、SP600125、SB239063与三叉神经节共同培养24 h,免疫组化和实时定量PCR法分别检测CGRP阳性反应细胞数和mRNA表达水平变化。结果:TG离体培养后12、24和48 h后CGRP阳性反应细胞数、平均吸光度和累积吸光度值均比新鲜组织明显增高,CGRP mRNA表达水平也明显增高。浓度为1×10^(-5)mol·L^(-1)的地塞米松、放线菌素D、环己酰胺以及U0126、SP600125分别与三叉神经节共同孵育24 h后,CGRP mRNA表达水平都显著降低。结论:TG离体培养后CGRP表达水平增高是通过细胞核内转录因子上调mRNA转录和蛋白翻译水平,是TG内非特异性炎症反应的结果,细胞内MAPK信号转导的ERK1/2通路和JNK信号通路参与了上调过程。Aim： To explore the intracellular signaling pathways involved in the up-regulation of calcitonin gene-related peptide（CGRP） in rat trigeminal ganglion （TG） after organ culture. Methods： Immunohistochemistry and real-time polymerase chain reaction （real-time PCR） were used to detect the CGRP positive cells and CGRP mRNA expression levels in TG after 12 h, 24 h and 48 h incubation with DMEM culture. The specific transcriptional inhibitor actinomycin D, the translational inhibitor cyclohexamide, the extracellular signal- regulated kinase 1/2 （ERK1/2） inhibitor U0126, protein kinase 38 （P38） inhibitor SB239063 and Jun N-terminal kinase （JNK） inhibitor SP600125, and non-specific inflammation inhibitor dexamethasone were respectively co-incubated with TG for 24 h. Results： The number of CGRP-positive cells and the levels of mRNA expression in TG significantly increased after culture for 12 h, 24 h and 48 h, compared with fresh TG tissue. The 1 ×10^-5 mol.Ll concentration of dexamethasone, cyclohexamide, actinomycin D, U0126 and SP600125 could remarkably decrease the up-regulated CGRP mRNA independently. Conclusion： The increasing level of CGRP in TG after organ culture was regulated by up-regulation of mRNA transcription and translation. It would be a non-specific intracellular inflammation process. The intracellular ERK 1/2 and JNK signal transduction pathway of MAPK was involved in CGRP up-regulation.

关 键 词：降钙素基因相关肽 三叉神经节 离体培养 信号转导 

分 类 号：R745.11[医药卫生—神经病学与精神病学]