HuD蛋白对肛门直肠畸形子鼠直肠末端肠神经系统的影响  

The impact of the HuD protein on the intestinal nervous system in the rectal end of animal models of congenital anorectal malformation

作　　者：孔萌[1,2] 刘远梅[2] 金祝[2] 高明娟[2] 曲颜[2] 郑泽兵[2] 

机构地区：[1]济南市儿童医院微创外科,250022 [2]遵义医学院附属医院小儿普胸泌外科,贵州563003

出　　处：《中华实验外科杂志》2016年第4期982-985,共4页Chinese Journal of Experimental Surgery

基　　金：基金项目：国家自然科学基金（81360067）;贵州省科技厅社发攻关项目[黔科合SY字（2009）3079]

摘　　要：目的检测HuD蛋白在肛门直肠畸形（ARM）子鼠直肠末端组织中的表达，探讨其可能对肠壁神经系统（ENS）发育的影响。方法选用20只成年sD孕鼠，随机分为正常组和乙烯硫脲（ETU）实验组各10只，两组孕鼠在孕20d取出宫内所有子鼠，观察其大体形态及计算畸形率，并取出子鼠直肠末端采用苏木素-伊红（HE）染色观察直肠末端组织结构变化，免疫组织化学和Westernblot检测HuD蛋白在直肠末端的表达。结果正常组共产子鼠108只，存活率为100．0％，未见任何畸形发生；实验组共产子鼠94只，存活率为93．6％，存活子鼠中ARM发生率为96．8％。HE结果显示：正常组子鼠直肠末端组织结构分界清楚，胞质丰富，轴突清晰；ETU组中患ARM的子鼠各层组织结构分层不清，可见较多空腔，直肠末端黏膜下和肌间神经元数目较正常组明显减少[（143．60±14．93）个比（335．90±10．46）个，P〈0．01]。免疫组织化学和Westernblot结果显示ARM组子鼠直肠末端蛋白含量明显低于正常组（312．90±53．40比456．40±57．13；0．24±0．05比0．45±0．06，P〈0．01）。结论HuD蛋白在ARM子鼠直肠末端表达差异有统计学意义，可能参与并影响ENS的发育与成熟。Objective To investigate the expression of the HuD protein in the rectal end of ano- rectal malformation （ARM） embryonic rats and to explore the effect of HuD expression on the development of the intestinal nervous system. Methods 20 adult pregnant Sprague Dawley rats were randomized into a control （ n = 10） and ARM （ induced by ethylenethiourea） group （ n = 10）. On the 20th day after pregnan- cy, the cesarean section fetus were obtained for morphological observations and abnormal statistics. Using hematoxylin - eosin （HE） staining to detect the perineum of sagittal gross specimen. Using immunohisto- chemistry （IHC） and Western blotting to detect the protein expression levels of HuD of distal rectum in fe- tal rats. Results The normal control group was given birth to 108 fetal rats whose survival rate was 100. 0% and no rectal abnormal openings, the ARM group was given birth to 94 fetal rats whose survival rate was 93.6% and ARM rate was 96. 8%. In HE staining, 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 hyperplas- tic squamous epithelial without intestinal gland distribution. The number of submucosal and myenteric plex- us alse reduced significantly in ARM fetuses compared with controls （ 143.60 ± 14.93 vs. 335.90 ± 10. 46, P 〈0. 01 ）. The immunohistochemistry and Western blotting analysis showed that the concentration of the HuD protein in the ARM group was significantly lower than that in the control group （ 312. 90 ± 53.40 vs. 456.40 ＋ 57.13 ; 0. 24 ＋ 0. 05 vs. 0. 45 ±0. 06, P 〈 0.01 ）. Conclusion The HuD protein was abnormally expressed in the rectal end of the ARM embryonic rats and may participate in the develop- ment and maturation of the enteric nervous system （ENS）. This study may provide a useful theoretical ref- erence for the treatment of postoperative defecation dysfunction in ARM patients.

关 键 词：肛门直肠畸形 肠神经系统 肌间神经元 HuD蛋白 

分 类 号：R726.571[医药卫生—儿科]