儿童原发性肾病综合征中血管生成素样蛋白3的表达  

Expression of ANGPTL3 in children with primary nephrotic syndrome

作　　者：饶佳[1] 徐虹[1] 孙利[1] 赵仲华[2] 张秀荣[2] 

机构地区：[1]复旦大学附属儿科医院肾科,上海200032 [2]复旦大学医学院病理学系

出　　处：《中华肾脏病杂志》2006年第5期286-290,共5页Chinese Journal of Nephrology

基　　金：上海市科委重点项目(034119809)

摘　　要：目的研究血管生成素样蛋白3(angiopoietin-like 3 protein,ANGPTL3)在儿童原发性肾病综合征(PNS)患者肾组织中表达分布及其参与蛋白尿发生的机制。方法 ANGPTL3分别与足细胞核标记抗原(WT1)、基底膜标记抗原类肝素硫酸蛋白多糖perlecan进行双标记法免疫荧光染色。应用免疫组化的方法检测ANGPTL3和pedecan在不同病理类型的69例PNS及血尿患儿,包括微小病变(MCD)31例、膜性肾病(MN)6例、局灶节段硬化性肾小球肾炎(FSGS)6 例、IgA肾病16例、薄基底膜肾病(TBMN)10例以及2例正常对照肾组织中表达,并以IMS彩色图像分析系统量化为免疫组化指数。在MCD病例中将尿蛋白肌酐比值分别与肾组织中 ANGPTL3和perlecan肾小球内染色强度及电镜下平均足突宽度(FWP)进行相关分析。对不同病理诊断时间(发病至肾穿刺)分组患儿肾小球ANGPTL3和perlecan的表达进行比较。结果 (1) ANGPTL3在正常肾组织呈现微弱的沉积,而在不同病理类型的肾病综合征患儿的肾组织的肾小球和肾小管存在不同程度的表达。肾小球内ANGPTL3表达量在MCD(7．49±1．96)、MN(6．27± 0．98)中显著高于正常对照(0．02±0．001)、TBMN(0．02±0．001)及FSGS(3．14±0．49)(均P< 0．05)。在IgA肾病(系膜增生型)中,蛋白尿组肾小球中ANGPTL3表达量显著高于单纯血尿组 (1．90±0．81比0．03±0．01,P<0．05)。(2)在MCD肾组织中,WT1及perlecan荧光双标记染色显示,ANGPTL3在足细胞胞浆及沿肾小球血管袢表达。(3)ANGPTL3在肾小球表达量分别与尿蛋白肌酐比值及电镜下平均足突宽度正相关(r为0．86、0．84,P均<0．05),并与perlecan在肾小球内表达量负相关(r为-0．83,P<0．05)。(4)不同发病年限的MCD患儿肾组织中肾小球 ANGPTL3及perlecan的表达无显著性差异。结论在不同程度的蛋白尿及不同足突融合程度的肾组织中存在ANGPTL3的表达差异。在MCD中,ANGPTL3主要在足细胞胞浆表达,肾小球中 Objective To examine the expression of angiopoietin-like protein（ANGPTL）3 in kidneys from children with primary nephrotic syndrome. Methods Immunohistochemistry for ANGPTL3 was performed in kidney biopsies from patients with nephrotic syndrome or hematuria, including MCD （n=31）, MN（n=6）, FSGS （n=6）, TBMN （n=10）, IgA nephropathy（IgAN） with mesangial proliferation （n=16）. Normal renal tissue of 2 cases with nephrectomy for tumor were used as control. According to the episode, four groups were divided （＂〈 1 month 1- 〈 6 months 6-12 months＂ and ＂〉12 months＂）. The expression was quantitatively examined with IMS color image analysis system, using positive index （PI） as sediment degree of ANGPTL3 in glomeruli or tubules. Immunofluorescence for ANGPTL3 co-labeling with WT1 and perlecan was applied to show the distribution of ANGPTL3. Results （1） The PI levels of ANGPTL3 in glomeruli of MCD（7.49± 1.96） and MN （6.27±0.98） were significandy higher than those of TBMN（0.02±0.001）, FSGS （3.14±0.49） or normal control（0.02±0.001） respectively （all P 〈 0.05）. The PI level of ANGPTL3 in glomeruli of IgAN patients accompanied with proteinuria was significantly higher than that of IgAN patients accompanied with hematuria （1.90±0.81 vs. 0.03±0.01, P 〈 0.05）. （2） Co-labeling with WT1, a marker of podocyte, showed that ANGPTL3 was expressed in the cytosol of WT1 positive cells under immunofluorescent microscope. Co-labeling with perlecan, a molecule secreted by podocyte and expressed in GBM, showed a strong positive selectively distribution at the periphery of glomerular capillary loops. （3） The PI level of ANGPTL3 in glomeruli was positively correlated with the ratio of urinary protein and creatinine , positively with F＇PW（r = 0.86, 0.84; P 〈 0.05）, and negatively with the PI level of porlecan in glomeruli （r = -0.83; P 〈 0.05）.（4） No difference was found among the four groups for the expression of ANGPTL3 nor porleca

关 键 词：血管生成素样类 肾病综合征 肾病 脂性 类肝素硫酸蛋白聚糖 

分 类 号：R726.91[医药卫生—儿科]