GDNF、P2Y2及HuD蛋白在肛门直肠畸形胎鼠及患儿直肠末端肠神经系统发育中的作用研究

先天性肛门直肠畸形(ARM)是小儿最常见的消化道畸形之一, 其发生原因不详，虽经手术治疗但仍因术后存在不同程度的排便障碍而影响患儿生活质量；近来研究发现可能与肠神经系统（ENS)发育异常有关。胶质细胞源性神经营养因子(GDNF)以及嘌呤受体亚型P2Y2和神经元标志物HuD蛋白与ENS发育密切相关，可能影响其排便功能。本课题应用乙烯硫脲制作大鼠不同胚胎时期ARM动物模型和收集高、中、低位ARM患儿手术的直肠末端标本,通过免疫组化、免疫荧光、western blot及qRT-PCR等方法观察GDNF、P2Y2、HuD在ARM不同时期胎鼠和不同类型ARM患儿直肠末端的表达情况，探讨GDNF、P2Y2以及HuD在ARM不同时期胎鼠及不同类型ARM患儿直肠末端肠神经系统发育中的作用，进一步明确GDNF、P2Y2、HuD蛋白与ARM排便功能障碍的关系，为临床治疗ARM患儿排便功能障碍奠定一定理论基础。

ARM（Anorectal Malformation）is one of the most common Children's digestive malformation and the cause is still unclear.Although ARM can be cured by surgery,some patients have disorders of defecation after surgery which affect the quality of children's life. Recently,it is researched that ARM may be related to the enteric nervous system（ENS）dysplasia. GDNF (glial cell line derived neurotrophic factor)、purine receptor subtype P2Y2 and neurons marker protein HuD are closely related to the development of ENS and these factors may affect bowel movement function.The intragastric administration of ethylenethiourea(ETU) is performed on fetal rats to create anorectal malformation model meanwhile different types of ARM rectum specimens will be collected by surgery.To study the expression of GDNF、P2Y2 and HuD in fetal rats rectum at different gestational age and children with different ARM types by the immunohistochemical/fluorescence、western blot and qRT-PCR. To explore the effects of GDNF、P2Y2 and HuD protein on the development of ENS between fetal rats in different gestational age and children with different ARM types，to further clarify the relationship between GDNF、P2Y2 and HuD protein and defecation dysfunction after ARM surgery,and to lay a certain theoretical basis about curing disorders of defecation ARM children.