The Expression And Significance Of P2Y₂ Purinergic Receptor Subtype In Hirschsprung's Disease

IntroductionHirschsprung's disease(Hirschsprung's disease,HD) is a common digestive ract of infant malformation,is a kind of Enteric nervous system(Enteric nervous system,ENS) dysphasia diseases.Neural crest-derived cells into the intestine gradually migration, proliferation,by the intestine microenvironmental impact to stop the proliferation,and gradually gathered to form the final differentiation of ganglion cells and glial cells, which constitute intestinal ganglion,in this process any process may lead to abnormal ganglion cells of the missing,and ultimately the formation of HD.As a neurotransmitter,Adenosine triphosphate(Adenosine triphosphate,ATP) may be involved in neuronal differentiation,migration,survival,but also promote neuronal repair,and its essential role with the receptor-binding,its receptor include P1 and P2,P2 receptors are divided into P2X,P2Y.So far there have been found eight subtypes of P2Y,in the gastrointestinal tract,the P2Y₂ subtype widely distributed.In the enteric nervous system P2Y receptor mainly regulate of muscle relaxation.In the development of enteric nervous system,if the expression of P2Y₂ deficiency may lead to bowel dysfunction of the relaxation,and thus reveals a certain relationship between the occurrence of HD and P2Y₂,but the reports about the expression and the distribution of P2Y₂ in HD is not yet see.In this study,we measured the expression of P2Y₂ receptor in children with normal,transitional and spasm paragraph.Normal paragraph intestine is set to in the control group,and transitional paragraphs,spasm as the experimental group land experimental group2,comparison the expression and distribution of P2Y₂ and explore its significance in the occurrence of HD.Materials and MethodsCase specimens Collected 30 cases of Surgical specimens in children with HD,children aged 23 days to 5.5 months,(25 cases of male,5 cases of female,the average age of 67.37±42.28 days)five cases of type long,short segment in 6 cases,common type of 19 cases.Reagents used for LaboratoryP2Y₂ receptor antibody(purchased from Abeam Inc.USA);two-step goat immuno-histochemical detection kit(purchased from Beijing Zhong Shan Golden Bridge Company);everse transcriptase kit(purchased from Takara Bio-engineering Co., Ltd.).Experimental methodsSelect the full-thickness organization of normal,transitional and spasm paragraph, were set to control group,experimental group land experimental group2,respectively. Using image analysis equipment to scan the results,and express the results using mean±standard,analysis the application with SPSS 13.0 software,inter-group differences using analysis of variance,the test results withp＜0.05 that have statistically significant.ResultThe results of immunohistochemical methodsIn the control group and experimental group 1,P2Y₂-positive cells are distributed and expressed in the longitudinal muscle,myenteric ring muscle and the nerve plexus. Compared with the control group,the number of P2Y₂-positive cells few and weak immune response in experimental group 1,the difference with statistical significance (p＜0.01).Experimental group 2 no obvious distribution and expression of P2Y₂-positive cells,compared with the control group the differences significantly(p＜0.01).The result of RT-PCR methodsP2Y₂ mRNA expressed in both control group(normal intestine) and experimental groupl(transitional paragraph intestinal),relative expression levels were(0.5736±0.07066),(0.2679±0.3817),the difference with statistical significance(p=0.000).There no obvious expression of P2Y₂ mRNA in experimental group2(spasm paragraph intestinal),there are differences between control group,experimental group land experi- mental group2(p value is 0.00,0.011). ConclusionCompared with the normal bowel paragraph,the expression of P2Y₂ receptor missing or significantly reduced in the intestinal lesions in children with HD,the deficiencies or significantly reduced of P2Y₂ expression may lead to bowel function of the relaxation,There is probably play an important role at the occurrence of HD, involved in the occurrence of HD.