PGP9.5在SD大鼠肠神经节细胞成熟度评价中的作用  

Protein gene product 9.5(PGP 9.5) in the evaluation of intestinal ganglion cells maturity in SD rats

作　　者：刘丽[1] 施诚仁[1] 张忠德[1] 李敏[2] 张文竹[2] 

机构地区：[1]上海第二医科大学附属新华医院上海儿童医学中心小儿外科,200092 [2]上海第二医科大学附属新华医院病理科,200092

出　　处：《中华小儿外科杂志》2003年第3期254-257,F003,共5页Chinese Journal of Pediatric Surgery

摘　　要：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 assess intestinal ganglion cells(IGCs) maturity in SD rats by immunohistochemical technique of PGP 9.5 and Cathepsin D and study the developmental events involved enteric nervous system (ENS).Methods Specimens from forty SD rats of different ages (10 for each group of E15, E20, P9 and adult rats) were stained conventionally with hematoxylin and eosin (HE), and immunohistochemically with PGP 9.5 and Cathepsin D. The size of positive IGCs was analyzed quantitatively.Results PGP 9.5 reliably stained the myenteric plexuses in all specimens and demonstrated the IGCs satisfactory when the latter were presented. While Cathepsin D expressed only in the brains of adult rats. The positive ratio of PGP 9.5 (100%) was higher than Cathepsin D (0%) in immature rats. The IGCs' size of E20 and P9 were smaller than that of adult rats ( P < 0.01 ).Conclusions PGP 9.5 may be used as a highly selective neuronal marker in the ENS, while the function of Cathepsin D should be studied further. Immunohistochemistry is a reliable simple technique for the maturity evaluation of intestinal ganglion cells and is useful in the study of congenital abnormality of the ENS.

关 键 词：先天性肠神经系统发育异常疾病 PGP9.5 免疫组织化学 神经元特异性烯醇化酶 

分 类 号：R726.5[医药卫生—儿科]