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目的 研究肠神经节细胞成熟度评价的方法和特异性抗体的作用 ,探讨PGP 9.5对研究先天性肠神经系统发育异常疾病的意义。方法 4 0只不同年龄的SD大鼠 :胎龄 15d(E15 ) ,2 0d(E2 0 ) ,生后 9d(P9) ,成年的SD大鼠各 10只。取标本 ,甲醛固定石蜡包埋切片 ,常规处理后用HE染色和PGP 9.5、CathepsinD、NSE的免疫组化染色 ,通过观察免疫染色阳性定位和定性情况和测量节细胞的平均最大直径 ,分析肠神经节细胞的发育和成熟状况。结果 PGP 9.5染色阳性率达 10 0 % ;NSE在E15中全部阴性 ,在E2 0和P9中皆呈弱阳性反应 ;而CathpsinD只在大脑中反应阳性。肠神经节细胞尺寸测量结果E2 0 ,P9和成年各年龄组间差异有显著性意义 (P <0 .0 1) ;PGP 9.5所显示的节细胞形成良好的成熟与幼稚的染色形态———成熟的节细胞胞浆颗粒清晰、细胞核淡染、核仁明显 ,未成熟的节细胞则胞浆胞核浓染、结构不清、无明显的核仁。结论 PGP 9.5在肠神经节细胞中的成熟与幼稚的不同染色形态可特征性地评价肠神经节细胞的成熟度。实验提示 ,PGP 9.5的免疫组化技术评价肠神经节细胞成熟度是一种可靠而简便的方法 ,在各种人类肠神经系统发育异常性疾病的形态学研究和临床诊治中有重要的实践意义。
Objective To study the method of evaluating the maturation of enteroglionic ganglion cells and the role of specific antibodies to explore the significance of PGP 9.5 in the study of congenital enteric nervous system dysplasia. Methods 40 male Sprague-Dawley rats of different ages were randomly divided into three groups: 10 days old (E15), 20 days old (E2 0), 9 days old (P9), and 10 adult SD rats. The specimens were fixed in formalin-fixed paraffin-embedded sections and stained with HE and PGP 9.5, Cathepsin D and NSE after routine treatment. The enteric nerve Cell development and maturation status. Results The positive rate of PGP 9.5 staining was 100%. NSE was negative in E15 and weakly positive in E2 0 and P9, while CathpsinD was only positive in brain. The results of the measurement of enteric ganglion cells showed significant difference between E2 0, P9 and adult age groups (P <0.01). PGP 9.5 cells showed well mature and naive staining patterns --- Mature cytoplasm of the cytoplasm particles clear, lightly stained nuclei, nucleoli obvious, immature section of the cytoplasm of nuclear stained cells, unclear structure, no obvious nucleolus. Conclusion The maturation of PGP 9.5 in gut ganglion cells and naive staining patterns can characteristically evaluate the maturation of enteric ganglion cells. The experimental results suggest that PGP 9.5 immunohistochemistry is a reliable and simple method to evaluate the maturation of enterogastric ganglion cells and has important practical significance in the morphological study and clinical diagnosis and treatment of various human intestinal disorders of the enteric nervous system .