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Preface congenital megacolon (Hirschsprung's disease, HD) is a common gastrointestinal infant deformities, is an enteric nervous system abnormalities (Enteric nervous system, ENS) disease. Neural crest-derived cells migrated into the intestine, proliferation, affected by intestinal microenvironment stop proliferation, gradually gathered eventually differentiate to form the ganglion cells and glial cells constitute the intestinal ganglion any exception in this process may lead to the absence of ganglion cells, and ultimately the formation of HD. Adenosine triphosphate (Adenosine triphosphate, ATP) as a neurotransmitter, may be involved in neuronal differentiation, migration, survival, and can also promote the repair of neurons play its role necessary for receptor binding, receptor P1 In two kinds P2, P2 receptors are divided into P2X, P2Y. P2Y 8 subtypes has been found so far, where the most widely distributed in the gastrointestinal tract P2Y 2 subtype. , P2Y receptors within the enteric nervous system is a major regulator of muscle relaxation, the missing relaxation may lead to bowel dysfunction If P2Y 2 expression in the enteric nervous system development process, so that the bowel is continuing spasticity, This reveals the P2Y 2 there is a certain relationship with the occurrence of HD, but the P2Y 2 HD intestinal expression and distribution of domestic not been reported. Through observation of HD in children with normal segment of the study, the transitional segment and spastic segment P2Y 2 receptor distribution and relative expression levels of the normal segment of the control group, transitional segments and spasm sections were set to experiment group 1 and experimental group 2, the comparison between the P2Y 2 distribution and expression of the existence of differences, to further explore the significance of this difference in HD pathogenesis. Materials and methods, case specimens collected from 30 patients with radical surgery of the China Medical University Shengjing Hospital Pediatric Surgery HD in children's surgical specimens, children aged 23 days (25 males and 5 females -5.5 The average age of 67.37 ± 42.28 days), including 5 cases of long segment short segment type in 6 cases, common type 19 cases. Second, experimental reagents P2Y 2 polyclonal antibody (purchased from Abcam); the two-step goat immunohistochemical detection kit (purchased from Wuhan Boster); reverse transcription kit (purchased from Takara Biotechnology Co., Ltd.). Experimental method to select children with normal intestinal specimens segment, transitional segment and spasm segment full thickness is set to the control group, the experimental group 1 and experimental group 2, P2Y observed by immunohistochemical method and RT-PCR method 2 distribution and expression in each group, all data are (?) ± s said that differences SPSS13.0 statistical software analysis was used to compare differences between groups using analysis of variance, p lt; 0.05 indicates statistical differences significance. Results immunohistochemical results in the control group and the experimental group, the longitudinal muscle layer of the intestinal specimens, ring muscularis and myenteric nerve plexus the visible P2Y 2 distribution of positive cells and expression, and control group compared to the experimental group P2Y 2 the number of positive cells and the immune response is weak, and the difference was statistically significant (P lt; 0.01). 2 intestinal specimens of the experimental group no the P2Y 2 -positive cell distribution and expression, significant differences (P lt; 0.01) compared with the control group. Second, RT-PCR experiments results in the control group (normal bowel segment) and the experimental group (transitional segment of intestine) in P2Y 2 mRNA expression relative expression level (0.5736 ± 0.07066), respectively, (0.2679 ± 0.3817), the difference was statistically significant (P = 0.000) mRNA the P2Y 2 in the experimental group 2 (spastic bowel segment) no significant expression in the control group, the experimental group The P2Y 2 expression in specimens with a significant difference (P is for 0.00,0.011). Conclusion tissue compared with normal bowel segment, HD children with lesions paragraph intestine in ATP receptor P2Y 2 expression missing or significantly reduce, the P2Y 2 expression deletion may cause intestinal relaxation of intestinal dysfunction in spasticity, may play a certain role in the occurrence of the process in HD.
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