CAJ | 학술논문

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,对照组灌入等量生理盐水,两组孕鼠在孕20 d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6.4％、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.
목적 관찰신경생장인자(NGF)급기수체TrkA、p75재선천성항문직장기형(ARM)동물모형태서직장말단적표체급분포.방법 선용성년SD잉서16지,실험조10지,대조조6지,실험조재잉11d급13d시분별이1％을희류뇨(125mg/kg)관위제작ARM동물모형,대조조관입등량생리염수,량조잉서재잉20 d수술취출궁내태서,진행형태학관찰급기형솔통계,병취태서분강、회음부시상면대체표본HE염색급직장말단석사포매절편,면역조화검측NGF급기수체TrkA、p75재정상태서화ARM태서직장말단적분포정황병반정량분석.결과 정상조:태서존활솔100％,항문개구정상,실험조:존활솔80.7％,실험조ARM발생솔98.7％、단미기형5.1％、무미기형94.8％、쌍족내번6.4％、척주렬6.4％,분강회음부시상절면HE염색현시:ARM태서항문위일폐합맹단,무정상항문결구,직장맹단위증생린상상피조직급증후적기층,미견장선분포,직장말단점막하급기간신경총수량:ARM조위46.40±16.08,대조조위96.33±18.57,량조비교,차이유통계학의의(P＜0.01);면역조화결과현시:NGF、TrkA、p75주요분포재태서직장말단점막하、기간신경총,기중NGF주요분포재세포핵,TrkA분포재세포질,p75재세포핵、세포질균유분포,기적분광밀도(IOD치):대조조분별위731.75±232.18、262.82±101.72、134.67±67.13,ARM조분별위17.97±74.95、33.15±11.13、113.01±36.47,량조비교,차이유통계학의의(P＜0.01).결론 NGF급기수체TrkA、p75가능공동삼여ARM적발생、발전과정.
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 pregnant 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 between the 11th and 13th day of gestation.On the 20th day of gestation,fetuses were delivered by cesarean section.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 studied 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 deformity(74.8％),varus feet(6.4％),and spinal bifida(6.4％).On the sagittal 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 imestinal glands distribution.The numbers of submucosal and myenteric plexus also reduced significantly in ARM fetuses 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 cytoplasm.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±232.18,262.82±101.72,134.67±67.13,P＜0.05).Conclusions The expressions of NGF and NTs receptors are decreased in the rectum of rat fetuses with ARM,which suggests NGF and NTs receptors may play an important role in the pathogenesis of ARM.