摘要
目的检测胶质细胞源性神经营养因子（GDNF）在肛门直肠畸形胎鼠直肠末端组织中的表达，探讨其可能与肠壁神经系统发育的关系。方法SD大鼠数字随机分对照组和乙烯硫脲致畸ARM组，两组孕鼠在孕20d手术取出宫内胎鼠，对胎鼠盆腔会阴部矢状面HE染色，免疫组化、Western blot方法检测对照组和ARM胎鼠直肠末端GDNF蛋白定位及定量表达情况，qRT-PCR方法检测两组GDNFmRNA的表达差异。结果HE染色显示：ARM胎鼠肛门为一闭合盲端，无正常肛门结构，直肠盲端为增生鳞状上皮组织及增厚的肌层，各层组织结构分层不清，直肠末端黏膜下和肌间神经丛数目较对照组明显减少（43．10±8．60比88．90±10．65，P〈0．01）。免疫组化结果显示GDNF在对照组及ARM组直肠末端黏膜层，黏膜下及肌间神经丛中均有表达，ARM组平均积分光密度（IOD值）明显低于正常组（238．32±41．67比500．93±90．10，P〈0．01）。Westernblot及qRT-PCR结果显示在ARM组胎鼠直肠末端组织中GDNF总蛋白及mRNA明显低于对照组（0．43±0．09比0．61±0．06；72．00±33．47比125．17±66．28，P〈0．05）。结论GDNF在肛门直肠畸形胎鼠直肠末端表达下调，可能与其肠壁神经系统发育不良有关。
Objective To explore the role of GDNF in the development of enteric nervous system of the anorectal Unit in fetal rats with malformations（ARM）. Methods The ARM was induced by ethylenethiourea（ETU） in fetal rats. On day 20 of gestation, fetal rats were harvested by cesarean section for HE staining of the perineum in sagittal section. The protein and RNA expression levels of GDNF were studied by irnmunohistochemistry, Western blot and qRT-PCR. Results HE staining showed no opening at the end of the distal rectum in ARM fetuses, and thickened muscular layer of the terminal rectum which covered by hyperplastic squamous epithelial without intestinal gland. The number of submucosal and myenteric plexus also reduced significantly in ARM fetuses compared with controis （43.10 ± 8. 60 vs 88. 90 ± 10. 65, P〈0. 01）. The expression of GDNF was mainly distributed in mucosal, submucosal plexus and myenteric plexus of the terminal rectum. The integral optical density （IOD）of GDNF was significantly lower in ARM fetuses than that of controls （238. 32 ± 41. 6750 vs 500. 93 ± 90./0, P〈0. 01）. The protein and mRNA expression of GDNF of ARM fetuses were 0. 43 ± 0. 09; 72. 00 ± 33.47, respectively, which were significantly lower than those of controls（0. 61 ± 0. 06, 125.17 ± 66. 28, P〈0. 05）. Conclusions The decreased expression of GDNF may play an important role in the maldevelopment of enteric nervous system in fetal rats with ARM.
出处
《中华小儿外科杂志》
CSCD
北大核心
2014年第1期62-65,共4页
Chinese Journal of Pediatric Surgery
基金
国家自然科学资金（81360067）
贵州省科技厅社发攻主项目[黔科合SY字（2009）3079]
关键词
胶质细胞源性神经营养因子
肠神经系统
直肠
Glial cell line-derived neurotrophic factor
Enteric nervous system
Rectum
