摘要
目的建立硝酸甘油诱发的大鼠偏头痛模型,观察大鼠硬脑膜上Artemin(Artn)的表达变化,探讨Artn在偏头痛病理发生的作用。方法 40只健康雌性Wistar大鼠随机分为模型组和生理盐水对照组。模型组采用颈部皮下注射硝酸甘油(10 mg/kg)诱发大鼠偏头痛模型,造模成功后0、2、4、6 h取硬脑膜检测。应用免疫荧光染色法定位Artn的表达;逆转录-聚合酶链反应(RT-PCR),Western blot检测Artn mRNA及蛋白的表达。结果双重免疫荧光结果显示Artn表达在偏头痛大鼠硬脑膜血管平滑肌细胞上,且注射硝酸甘油6 h后Artn的免疫阳性信号与生理盐水对照组相比明显增强。生理盐水对照组各时间点均可见Artn的表达,但各时间点的相对表达差异无统计学意义(P>0.05)。模型组Artn mRNA在2 h时表达量达高峰(1.520±0.048),与生理盐水对照组相比(0.474±0.026),差异具有统计学意义(t2h=33.247,P<0.01)。随后表达量逐步下降,6 h趋于正常对照组水平。模型组Artn的蛋白在0和2 h时与生理盐水对照组相比表达量无变化,而4和6 h表达量迅速增加,差异具有统计学意义(t4h=7.208,P4h<0.05;t6h=16.491,P6h<0.01)。结论 Artn可能通过其表达量的上调参与了偏头痛的病理发生过程。
OBJECTIVE To observe the expression of Artemin（Artn）in migrainous rat dura mater induced by nitroglycerin（NTG）and to investigate the possible role of Artn in the pathogenesis of migraine.METHODSForty Adult female Wistar rats were randomly divided into control group and model group.The migraine rat model was triggered by subcutaneous injection of NTG（10 mg/kg）while the counterpart with normal saline（NS）.After the model was set up,the dura mater was removed at 0,2,4,6 h and the expressions of Artn were investigated by RT-PCR,Western-blot,and double immunofluorescence labeling.RESULTS Artn was expressed in smooth muscle cells of dural vasculature in a model of migraine and the expression of Artn conspicuously increased at 6 h after injection of NTG compared to injection of NS.The positive expression of Artn was detected at all indicated time points in the control groups,but there were no significant differences at all time points（P〈0.05）.Compared with injection of NS,the mRNA expression of Artn reached its peak level at 2 h after injection of NTG（1.520±0.048 vs.0.474±0.026,P〈0.01）,here after,the expression of the mRNA of Artn declined and returned to the level of the control group at 6 h after NTG injection.The protein expressions of Artn in NTG group at 0 h and 1 h were not different from that of control group,but markedly increased at 4 h and 6 h.CONCLUSION The up-regulated expression of Artn may play an important role in the pathogenesis of migraine.
出处
《中国耳鼻咽喉头颈外科》
北大核心
2011年第12期663-666,共4页
Chinese Archives of Otolaryngology-Head and Neck Surgery
基金
国家自然科学基金项目(81170907)
山东省科技攻关计划(2007GG20002041)联合资助
关键词
动物实验
偏头痛
硝酸甘油
模型
动物
Animal Experimentation
Migraine Disorders
Nitroglycerin
Models
Animal
