CAJ | 학술논문

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GDNF mRNA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).Western blot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.
목적 검측효질세포원성신경영양인자(GDNF)재항문직장기형태서직장말단조직중적표체,탐토기가능여장벽신경계통발육적관계.방법 SD대서수자수궤분대조조화을희류뇨치기ARM조,량조잉서재잉20d수술취출궁내태서,대태서분강회음부시상면HE염색,면역조화、Western blot방법검측대조조화ARM태서직장말단GDNF단백정위급정량표체정황,qRT-PCR방법검측량조GDNF mRNA적표체차이.결과 HE염색현시:ARM태서항문위일폐합맹단,무정상항문결구,직장맹단위증생린상상피조직급증후적기층,각층조직결구분층불청,직장말단점막하화기간신경총수목교대조조명현감소(43.10±8.60비88.90±10.65,P＜0.01).면역조화결과현시GDNF재대조조급ARM조직장말단점막층,점막하급기간신경총중균유표체,ARM조평균적분광밀도(IOD치)명현저우정상조(238.32±41.67비500.93±90.10,P＜0.01).Western blot급qRT-PCR결과현시재ARM조태서직장말단조직중GDNF총단백급mRNA명현저우대조조(0.43±0.09비0.61±0.06；72.00±33.47비125.17±66.28,P＜0.05).결론 GDNF재항문직장기형태서직장말단표체하조,가능여기장벽신경계통발육불량유관.
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 immunohistochemistry,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 controls (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 ± 90.10,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.