甘氨酸受体β亚基蛋白在慢性胰腺炎疼痛模型中的表达及意义

Expression and significance of glycine receptor beta subunit in the model of chronic pancreatitis for pain study

目的 观察抑制性神经递质甘氨酸受体β亚基（GLRB）在慢性胰腺炎（CP）疼痛模型中的表达,探讨其与CP炎疼痛发生的关系.方法 30只雄性Wistar大鼠随机分成正常对照组、磷酸盐缓冲液（PBS）胰管内注射假手术组、三硝基苯磺酸（TNBS）胰管内注射模型组3组,通过胰腺组织苏木素-伊红（HE）染色及疼痛阈值测定判定造模成功与否.荧光实时定量聚合酶链反应（FQ-PCR）检测脊髓背根神经节中GLRB基因表达水平,免疫组织化学检测其表达部位及强弱.根据GLRB表达水平分析其与疼痛相关性.结果 造模前后,3组疼痛阈值测定均成动态变化,先降低后升高,造模第4周,造模组疼痛阈值低于假手术组和对照组（3组比较：x2=4.279＜x20.05（8）,P＞0.05;两两对比：P ＜0.05）.在GLRB基因表达层面上,造模组较假手术组和对照组表达上调,造模组GLRB相对表达量为4.545 2±1.886 0,假手术组GLRB相对表达量为1.841 0±1.0490,对照组GLRB相对表达量为1.4920±1.1370（上调约2.5倍,P＜0.05）.免疫组织化学结果示表达部位主要于神经节细胞胞质.结论 抑制性神经递质GLRB在CP脊髓背根神经节组织中表达增加,疼痛阈值呈先降低后升高至接近正常的动态变化,可能是自我保护机制的负反馈调节作用.

Objective To study the expression of glycine receptor beta subunit in the model of chronic pancreatitis,and evaluate the relationship between glycine receptor beta subunit and pain in chronic pancreatitis.Methods Thirty male Wistar rats were randomly divided into three groups：normal group,phosphate buffer （PBS） perfusion group,and trinitrobenzene sulfonic acid perfusion group.Histopathological and pain examinations were done to confirm the establishment of chronic pancreatitis model.Real-time fluorescent quantitative polymerase chain reaction （FQ-PCR） was applied to detect the expression of glycine receptor beta subunit.Immunohistochemistry was used to localize the expression site of glycine receptor beta subunit.Results Pain examination in this model showed dynamic,decrease at first,increase at last.Compared with another groups,the trinitrobenzene sulfonic acid perfusion group is lower in the aspect about the pain during the fourth week （comparing the three groups：x2 =4.279〈x0.05 （8）,P 〉 0.05 ; comparing with each other：P 〈0.05）,but higher than others in gene expression （2.5 times,P 〈0.05）,and strongly expression was observed in cytoplasm.Conclusion Enhanced expression of glycine receptor beta subunit,the inhibitory neurotransmitter,is involved in the pain of chronic pancreatitis.For self-protection,it reduces the degree of pain below normal lever.