胶质细胞系源性神经营养因子/胶质细胞系源性神经营养因子受体信号通路在乳鼠先天性巨结肠相关性小肠结肠炎中的作用实验研究  

Role of GDNF/GFRα1 signaling pathway in neonatal rats with Hirschsprung-associated enterocolitis

作　　者：牛志新 何峰 宋春光[1] 肖玉清 NIU Zhixin;HE Feng;SONG Chunguang;XIAO Yuqing(Department of Proctology,the First Hospital of Qinhuangdao,Qinhuangdao 066000,China)

机构地区：[1]秦皇岛市第一医院肛肠科,河北秦皇岛066000 [2]青龙满族自治县医院外二科,河北秦皇岛066500

出　　处：《陕西医学杂志》2024年第1期19-22,共4页Shaanxi Medical Journal

基　　金：河北省自然科学基金资助项目(H2021206217)。

摘　　要：目的:探讨胶质细胞系源性神经营养因子/胶质细胞系源性神经营养因子受体(GDNF/GFRα1)通路在乳鼠先天性巨结肠(HD)相关性小肠结肠炎(HAEC)中的作用研究。方法:21日龄内皮素受体(Ednrb)基因敲除乳鼠为HD模型组(实验组),同日龄野生型乳鼠为对照组,8只/组。取结肠组织,按照形态将实验组乳鼠结肠组织分为狭窄段、扩张段,对照组乳鼠结肠组织分为远段、近段。采用苏木精-伊红(HE)染色法评判两组肠管炎症程度;酶联免疫吸附法(ELISA)法检测两组结肠组织白介素-10(IL-10)、肿瘤坏死因子-α(TNF-α)水平;免疫印迹法(WB)分别检测GDNF/GFRα1信号通路相关基因[GDNF、GFRα1、酪氨酸酶受体(RET)、核转录因子kappaBp65(NF-κBp65)、TNF-α]蛋白水平。结果:HE染色结果显示HAEC主要发生在结肠扩张段;ELISA法结果显示,与实验组乳鼠狭窄段肠管比较,扩张段肠管IL-10水平显著降低、TNF-α水平显著升高(均P<0.05);WB法结果显示,与实验组乳鼠狭窄段肠管比较,扩张段肠管GDNF、GFRα1、RET蛋白水平显著降低,NF-κBp65、TNF-α蛋白水平显著升高(均P<0.05)。结论:HD模型乳鼠结肠扩张段炎症程度较为严重,机制可能与GDNF/GFRα1信号通路受抑制,肠神经免疫调节系统异常有关。Objective:To investigate the role of glial cell line derived neurotrophic factor/glial cell line-derived neurotrophic factor receptor(GDNF/GFRα1)pathway in neonatal rats with Hirschsprung-associated enterocolitis(HAEC).Methods:The 21-day-old Ednrb knockout mice were HD model group(experimental group)and the same-day-old wild-type mice were control group,8 mice in each group.According to the morphology of colon tissue,the colon tissue of the experimental group was divided into narrow segment and dilated segment,and the colon tissue of the control group was divided into distal segment and proximal segment.The degree of intestinal inflammation was evaluated by hematoxylin-eosin(HE)staining,and the levels of interleukin-10(IL-10)and tumor necrosis factor(TNF-α)were measured by Elisa staining GDNF/GFRα1 signaling pathway related genes(GDNF,GFRα1,tyrosine kinase receptor[RET],nuclear transcription factorκB p65[NF-κB p65]and TNF-α)were detected by Western blot(WB).Results:HE staining showed that HAEC mainly occurred in the dilated segments of the colon.ELISA results showed that compared with the narrow segment of the intestines in the experimental group,the level of IL-10 in the dilated segment of the intestines was significantly decreased,and the level of TNF-αwas significantly increased(both P<0.05).The results of WB showed that compared with the narrow intestinal segment in the experimental group,the protein levels of GDNF,GFRα1 and RET in the dilated intestinal segment in the experimental group were significantly decreased,and the protein levels of NF-κB p65 and TNF-αwere significantly increased(all P<0.05).Conclusion:Inflammation in the dilated colon of HD mice is severe,which may be related to the inhibition of GDNF/GFRα1 signaling pathway and the abnormality of the intestinal neuroimmunoregulatory system.

关 键 词：胶质细胞系源性神经营养因子 胶质细胞系源性神经营养因子受体通路 先天性巨结肠 相关性小肠结肠炎 肿瘤坏死因子-α 核转录因子κBp65 

分 类 号：R574.62[医药卫生—消化系统]