【摘要】 目的免疫组织化学方法检测4种肠道神经标志物在正常结肠、先天性巨结肠症(HD)、神经元发育不良-B(IND-B)及神经节细胞减少症(HG)病变结肠的表达特点,并采用图像分析技术对肠道神经系统进行定量分析。方法组织蛋白酶D、蛋白基因产物9.5、S-100蛋白和外周蛋白4种抗体,对10例正常结肠、20例HD、8例IND-B型及20例HG的患儿结肠石蜡标本切片采用过氧化酶标记的链霉卵白素免疫组织化学染色。ImageJ图像处理软件对所有数码图像(×400)中神经丛、节细胞计数以及节细胞形态进行定量测量。结果组织蛋白酶D仅在神经节细胞表达阳性,不表达于神经纤维;PGP 9.5和外周蛋白在肠壁肌间、黏膜下神经丛及神经节细胞中均有阳性表达;S-100在神经节细胞中表达阴性。定量分析:HD病变肠段缺乏神经节细胞;IND-B病变肠段肌间神经节细胞与正常比较计数明显增多,节细胞直径以及细胞核直径分别为正常的1.22倍和1.34倍(P<0.01);HG病变肠段神经丛内节细胞数量明显减少(P<0.05)。结论免疫组织化学染色图像分析技术使不同病变间的形态差异得到定量分析,有助于对HD和同源病的病理学研究以及临床诊断和鉴别诊断。更多还原

【Abstract】 Objective To investigate the diagnostic role of 4 enteral neuronal markers in Hir- schsprung’s disease(HD)and its allied disorders(AHDs)(Intestinal Neuronal Dysplasia-B(IND-B) and Hypoganglionosis(HG)).Methods According to pathological diagnosis,intestinal specimens were divided into four groups;normal 10,HD 2(1,IND 8 and HG 20 cases.Specimens were paraffin embedded and cut into 5-μm-thick slices.Tissue slices were stained immunohistochemically with Streptavidin/Peroxidase(SP),using Catepsins D,PGP 9.5,S-100 and Peripherin antibodies.The components of ENS in digital images(×400)were measured with Image J software(NIH).Results The staining characteristics in normal ENS were;Catepsins D only presented in ganglion cell,but didn’t expressed in nerve plexus;the expressions of PGP 9.5 and Peripherin could he shown in sub- mucosal plexus,myenteric plexus and in the ganglion cells;S-100 expressed in submucosaI and myen- teric plexus,but didn’t express within ganglion cells.Quantitative analysis for HD and AHD;few ganglion cells could be found in nerve plexus of HD;only 1-2 ganglion cells could be detected in plexus of HG;the ganglion cells were increased markedly in IND.Comparing with normal group,cell and nu- cleus diameter of ganglion were obviously reduced in HG,but were increased in IND(P<0.01).Con- clusions Image analysis of immunohistochemical staining enables us to analyze accurately morphologi- cal difference between the different pathological changes in HD and AHDs.更多还原