先天性巨结肠患儿肠壁中转录因子NKX2-1表达的研究

发布时间：2018-06-20 18:43

本文选题：先天性巨结肠 + 转录因子NKX2-1　；参考：《郑州大学》2012年硕士论文

【摘要】：背景和目的先天性巨结肠症又称为赫希施普龙病(Hirschsprung's disease,HD)是一种比较常见的小儿消化道发育畸形,属于一种先天性疾病。发病率约为1/5000,存在一定的性别差异,男女比例约为4：1。HD是造成结肠肠管增粗的原因之一可发生在近端结肠,主要位于乙状结肠和部分降结肠；也可发生在直肠和远端乙状结肠；HD患儿结肠狭窄段和扩张段两部分结肠组织之间有一过渡或移形区,往往呈漏斗形,长约3cm-8cm不等。已有实验证实了转录因子NKX2-1基因蛋白在整个肠神经系统的发育中存在。近年来NKX2-1的研究主要集中在甲状腺、脑及肺组织等有关神经系统发育疾病过程的研究中,在肠神经系统的研究比较少。先天性巨结肠患儿的肠蠕动功能可能与转录因子NKX2-1基因蛋白在HD患儿肠管管壁中的表达存在一定的相关性,本次实验的目的是应用免疫组化SP法和逆转录聚合酶链反应(RT-PCR)技术定性和定量测定转录因子NKX2-1基因在HD患儿结肠壁中的表达,以便更加深入的了解HD在分子基础上的确切发病机制,为HD疾病的临床诊断提供可靠的参考。材料和方法 1应用免疫组化SP法观察30例HD手术切除标本(分别取HD患儿狭窄段、移行段、扩张段结肠壁全层组织)同时取30例肠套叠患儿手术切除并具备神经节细胞的全层结肠组织做为对照组,观察各组织标本中转录因子NKX2-1基因在HD患儿结肠壁中的表达情况； 2应用逆转录聚合酶链反应方法检测30例新鲜的HD手术切除标本(包括HD患儿狭窄段、移行段、扩张段全层结肠组织)和30例肠套叠患儿手术切除的存在神经节细胞的全层结肠组织做对照组,观察转录因子NKX2-1基因在HD患儿结肠组织中的表达情况； 3统计学处理：数据应用SPSS17.0软件处理,HD患儿各组结肠组织和正常对照组结肠组织中转录因子NKX2-1基因蛋白表达的比较采用两个独立样本的秩和检验；HD患儿各组结肠组织和正常对照组结肠组织中转录因子NKX2-1mRNA表达的比较采用两个独立样本的t检验或校正t检验。检验水准a=0.05。结果 1转录因子NKX2-1在巨结肠患儿窄段结肠组织中表达明显减少,在扩张段结肠组织中表达正常,在移行段中的表达处于二者之间。 2HD患儿狭窄、移行段结肠组织中转录因子NKX2-1基因蛋白和nRNA表达与正常对照组结肠组织中转录因子NKX2-1基因蛋白的mRNA表达进行比较,差异有统计学意义(Z=5.236和2.926,P0.008；t’=19.475和14.429,P0.05),HD患儿扩张段结肠组织和对照正常结肠组织中NKX2-1蛋白和mRNA的表达比较,差异无统计学意义(Z=2.336,P=0.0190.008；t=0.985,P=0.3290.05)。结论 1HD患儿狭窄段结肠壁组织中缺少转录因子NKX2-1基因的表达是HD发病的基本病理改变之一。 2转录因子NKX2-1基因在HD患儿结肠组织中表达减少是导致HD发病的原因之一。
[Abstract]:Background and objective Hirschsprung disease (HD), also known as Hirschsprung disease (HD), is a common developmental malformation of digestive tract in children and belongs to a congenital disease. The incidence rate is about 1 / 5 000, there is a certain sex difference, the ratio of male and female is about 4: 1. HD is one of the causes of colon bowel thickening, mainly located in sigmoid colon and part descending colon. It may also occur in children with rectum and distal sigmoid colon HD. There is a transitional or shifting area between the stricture and dilation of colon in children with rectum and distal sigmoid colon. It is usually funnel-shaped, and the length of the colon varies from that of 3cm-8cm. It has been confirmed that NKX 2-1 gene protein exists in the development of the enteric nervous system. In recent years, the study of NKX2-1 is mainly focused on thyroid, brain and lung tissues and other related neurological diseases, but rarely on intestinal nervous system. The intestinal peristalsis function in children with Hirschsprung's disease may be related to the expression of NKX2-1 gene protein in the intestinal wall of HD children. The aim of this study was to determine the expression of NKX2-1 gene in colon wall of HD children by immunohistochemical SP method and reverse transcriptase polymerase chain reaction (RT-PCR). In order to further understand the molecular basis of HD pathogenesis and provide a reliable reference for the clinical diagnosis of HD disease. Materials and methods 1 Immunohistochemical SP method was used to observe 30 HD surgical specimens. 30 patients with intussusception were selected as control group. The expression of NKX2-1 gene in colon wall of HD patients was observed. 2 reverse transcriptase polymerase chain reaction (RT PCR) was used to detect the stenosis and transitional segments of 30 fresh HD surgical specimens (including HD children). The expression of NKX2-1 gene in colon tissue of HD children was observed by observing the expression of NKX2-1 gene in colon tissue of 30 children with intussusception. 3Statistical processing: the expression of NKX2-1 gene protein in colon tissue of HD children was analyzed by SPSS 17.0 software and the expression of NKX2-1 gene protein in colon tissue of HD group was compared with that of normal control group. Two independent samples were used to test the expression of NKX2-1 gene protein. The expression of transcription factor NKX2-1 mRNA in colon tissue of HD children was compared with that in normal control group. The expression of NKX2-1 mRNA in colon tissues of two independent samples was examined by t-test or corrected t-test. The inspection level is 0. 05. Results (1) the expression of NKX2-1 in the narrow segment of Hirschsprung's colon was significantly decreased, the expression of NKX2-1 was normal in the dilated colon, and the expression of NKX2-1 in the transitional segment was between the two. The expression of NKX2-1 gene protein and nRNA in the transitional colon tissue was compared with that in the normal colon tissue. The difference was statistically significant (P 0.008) between the expression of NKX2-1 gene protein and the mRNA expression of NKX2-1 gene protein in the transitional colon tissue. There was no significant difference in the expression of NKX2-1 protein and mRNA between dilated colon tissue and normal colon tissue of children with Tet (19.475) and 14.429 (P 0.05). There was no significant difference in the expression of NKX2-1 protein and mRNA between the two groups. There was no significant difference in the expression of NKX2-1 protein and mRNA between dilated colon tissue and normal colon tissue. There was no significant difference in the expression of NKX2-1 protein and mRNA between the two groups. Conclusion 1 the lack of expression of NKX2-1 gene is one of the basic pathological changes of HD. 2 the expression of NKX2-1 gene decreases in colon tissue of HD children. Is one of the causes of HD.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R726.5

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