先天性肛门直肠畸形胎鼠直肠末端NGF及受体TrkA与p75的表达  

The expressions of NGF and NTs receptors （TrkA and p75） in distal rectum of fetal rats with anorectal malformations

作　　者：毛羽晨[1] 刘远梅[1] 金祝[1] 高明娟[1] 郑泽兵[1] 周涛[1] 

机构地区：[1]遵义医学院附属小儿普胸泌科,563003

出　　处：《中华小儿外科杂志》2012年第11期854-858,共5页Chinese Journal of Pediatric Surgery

基　　金：贵州省科技厅社发公关项目[黔科合SY字（2009）3079]

摘　　要：目的观察神经生长因子（NGF）及其受体TrkA、p75在先天性肛门直肠畸形（ARM）动物模型胎鼠直肠末端的表达及分布。方法选用成年SD孕鼠16只，实验组10只，对照组6只，实验组在孕11d及13d时分别以1％乙烯硫脲（125mg／kg）灌胃制作ARM动物模型，对照组灌人等量生理盐水，两组孕鼠在孕20d手术取出宫内胎鼠，进行形态学观察及畸形率统计，并取胎鼠盆腔、会阴部矢状面大体标本HE染色及直肠末端石蜡包埋切片，免疫组化检测NGF及其受体TrkA、p75在正常胎鼠和ARM胎鼠直肠末端的分布情况并半定量分析。结果正常组：胎鼠存活率100％，肛门开口正常，实验组：存活率80．7％，实验组ARM发生率98．7％、短尾畸形5．1％、无尾畸形94．8％、双足内翻6．4oA、脊柱裂6．4％，盆腔会阴部矢状切面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，未见肠腺分布，直肠末端黏膜下及肌间神经丛数量：ARM组为46．40±16．08，对照组为96．33±18．57，两组比较，差异有统计学意义（P〈0．01）；免疫组化结果显示：NGF、TrkA、p75主要分布在胎鼠直肠末端黏膜下、肌间神经丛，其中NGF主要分布在细胞核，TrkA分布在细胞质，p75在细胞核、细胞质均有分布，其积分光密度（IOD值）：对照组分别为731．75±232．18、262．82±101．72、134．67±67．13，ARM组分别为17．97±74．95、33．15±11．13、113．01±36．47，两组比较，差异有统计学意义（P〈0．01）。结论NGF及其受体TrkA、p75可能共同参与ARM的发生、发展过程。Objective To study the expressions of nerve growth factor （NGF） and NTs receptors （TrkA and p75） in distal rectum of fetal rats with anorectal malformations （ARM）. Methods Sixteen preg- nant SD rats were divided into ARM group （n=10） and normal control group （n=6）. Ethylenethiourea was administered to the pregnant rats by adding to drinking water to induce anorectal malformation in fetus be-tween the 11th and 13th day of gestation. On the 20th day of gestation, fetuses were delivered by cesarean sectiorL The fetuses were sectioned serially and sagittally, morphology of the distal rectum was studied. The expressions and distribution of NGF,TrkA and p75 in distal rectum and peritoneum of the fetuses were stud-ied using immunohistochemical staining. Results The fetuses of the control group all survived after cesarean section and no abnormal rectal openings were found. The survival rate of the fetuses of ARM group was 80. 7%. And ARM was found in 98. 7% of the fetuses. Other deformities were also noted including short-tailed deformity （25. 1%） ,no tail defomaity （74. 8%） ,varus feet （6. 4%） ,and spinal bifida （6. 4%）. On the sagit-tal section of peritoneum, there was no opening at the end of the distal rectum of the ARM fetuses, the end of the rectum had thickened muscular layer and covered by hyperplastic squamous epithelium without intestinal glands distribution. The numbers of submucosal and myenteric plexus also reduced significantly in ARM fetu-ses compared with controls （46. 40 ± 16. 08 vs 96. 33 ± 18. 5, P〈0. 01 ）. The NGF, TrkA, and p75-positive cells were mainly distributed in the terminal rectum mucosa and myenteric plexus. NGF protein was localized mainly in nucleus. TrkA protein was localized mainly in cytoplasm and p75 protein in nucleus and cytoplasrm. The integral optical densities （IOD） of NGF,TrkA,and p75 of ARM fetuses were 17. 97 ± 74. 95,33. 15 ± 11. 13,113.01 ± 36. 47,respectively,which were significantly lower than those of controls （731.75 ±

关 键 词：肛门 畸形 直肠 畸形 神经生长因子 

分 类 号：R725.7[医药卫生—儿科]