嘌呤受体亚型(P2Y2)在肛门直肠畸形动物模型直肠末端的表达
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摘要
目的检测嘌呤受体亚型(P2Y2)在肛门直肠畸形(ARM)胎鼠直肠末端的表达情况,探讨其可能对肠壁神经系统发育的影响。方法 SD孕鼠随机分正常组和乙烯硫脲(ETU)实验组,两组孕鼠在孕20d取出宫内胎鼠直肠末端,应用HE染色观察直肠末端组织结构变化,Western blot和qRT-PCR方法检测P2Y2在直肠末端的表达情况。结果正常组胎鼠直肠末端组织结构分界清楚,黏膜下和肌间神经元细胞数目多,胞浆丰富,轴突清晰;ETU组中患ARM的胎鼠各层组织结构分层不清,可见较多空腔,神经元细胞数目较少。Western blot和qRT-PCR结果显示ARM组胎鼠直肠末端蛋白含量及mRNA量明显低于正常组(0.23±0.06比0.42±0.05;49.40±26.49比98.53±52.25,P<0.05)。结论 P2Y2可能参与ARM胎鼠直肠末端肠神经系统的发育。
Objective To detect the expression of P2Y2 in anorectal malformation(ARM) in fetal rat distal rectum and explore its possible significance.Methods Adult pregnant SD rats were randomly divided into normal control group(NG) and ARM group induced by ethylenethiourea(ETU) in fetal rats.On the 20th day after pregnancy,distal rectum tissues of the fetal rats were collected from the two groups and stained by HE staining.The expression of P2Y2 terminal rectum was detected by Western blot and qRT-PCR assay.Results The terminal rectum tissues,submucosal and myenteric neurons number,abundant cytoplasm and axon were clear in NG group.However,each layer hierarchical structure was not clear,visible more cavity and the number of neurons was less in the ARM group.Western blot and qRT-PCR results showed that the expression of P2Y2 terminal rectum in ARM group is significantly lower than those in NG group(0.23 ± 0.06 vs 0.42 ± 0.05;49.40 ± 26.49vs 98.53 ± 52.25,P < 0.05).Conclusion P2Y2 might be involved in the development of enteric nervous system in ARM rats.
Objective To detect the expression of P2Y2 in anorectal malformation(ARM) in fetal rat distal rectum and explore its possible significance.Methods Adult pregnant SD rats were randomly divided into normal control group(NG) and ARM group induced by ethylenethiourea(ETU) in fetal rats.On the 20th day after pregnancy,distal rectum tissues of the fetal rats were collected from the two groups and stained by HE staining.The expression of P2Y2 terminal rectum was detected by Western blot and qRT-PCR assay.Results The terminal rectum tissues,submucosal and myenteric neurons number,abundant cytoplasm and axon were clear in NG group.However,each layer hierarchical structure was not clear,visible more cavity and the number of neurons was less in the ARM group.Western blot and qRT-PCR results showed that the expression of P2Y2 terminal rectum in ARM group is significantly lower than those in NG group(0.23 ± 0.06 vs 0.42 ± 0.05;49.40 ± 26.49vs 98.53 ± 52.25,P < 0.05).Conclusion P2Y2 might be involved in the development of enteric nervous system in ARM rats.
引文
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[12]Donnell A M,Purl P.Deficiency of purinergic P2Y2 receptors in aganglionic intestine in Hirsehsprung’s[J].Pediatr Surg Int,2008,24(1):77-80.
[2]Wa H,Hughes I,Planer W,et al.The timing and location of glial cell line-derivedneurotrophic factor expression determine enteric nervous system structure and function[J].Neurosci Res,2010,30(4):23-38.
[3]Young H M,Newgreen D.Enteric neural crest-derived cells:Origin,identification,and differentiation[J].Anat Rec,2001,262(1):12-15.
[4]Xiang Z,Bumstock G.Distribution of P2Y2 receptors in theguinea pig enteric nervous system and its coexistence with P2X2 and P2X3 receptors,neuropeptide Y,nitric oxide synthase and ealretinin[J].Histochem Cell Biol,2005,124(5):379-390.
[5]刘远梅,毛羽晨,金祝,等.神经生长因子及其受体在肛门直肠畸形动物模型直肠末端的表达及意义[J].中华小儿外科杂志,2011,32(12):924-927.
[6]陈兰萍,陈新新,任红霞.先天性肛门直肠畸形患儿术后排便功能障碍病因研究进展[J].中华小儿外科杂志,2012,33(7):541-543.
[7]毛羽晨,刘远梅,金祝,等.先天性肛门直肠畸形胎鼠直肠末端NGF及受体TrkA与P75的表达[J].中华小儿外科杂志,2012,33(11):854-858.
[8]Ralevic V,Burnstock G.Receptors for purines and pyrimidines[J].Harmacol Rev,1998,50(3):413-492.
[9]Arthur D B,Akassoglou K.P2Y2 receptor activates nerve growth actor/TrkA signaling to enhance neuronal differentiation[J].Proc Natl Acad Sci USA,2005,102(52):19138-19143.
[10]Thornton P D,Bornstein J C.Slow excitatory synaptic potentials evoked by distension in myenteric descending interneurones of guinea pig ileum[J].Physiol,2002,539(Pt 2):589-602.
[11]Agresti C,Meomartini M E,Amadio S,et al.ATP regulates oligodendrocyte progenitor migration,proliferation,and differentiation:involvement of etab-otropic P2 receptors[J].Brain Res Rev,2005,48(2):157-165.
[12]Donnell A M,Purl P.Deficiency of purinergic P2Y2 receptors in aganglionic intestine in Hirsehsprung’s[J].Pediatr Surg Int,2008,24(1):77-80.