IgA肾病系膜细胞-足细胞轴在肾小管间质病变中的作用

目的探讨IgA肾病系膜细胞-足细胞轴在肾小管间质病变中的作用。方法收集40例经肾穿刺活检明确诊断的IgA肾病,采用免疫组化法检测足细胞表面特异性标志物WT-1、肾组织TNF-α、IL-6的表达;采用酶联免疫吸附法测定患者尿TNF-α、IL-6的活性水平。结果 IgA肾病患者肾小球系膜增生评分与WT-1的表达呈负相关(r=-0.763,P0.05),与肾组织TNF-α及IL-6表达、尿液TNF-α及IL-6活性水平均呈正相关(r=0.674、r=0.853、r=0.819、r=0.763,P均0.05),IgA肾病患者随肾小管间质病变加重,肾组织WT-1的表达逐渐降低;而TNF-α、IL-6的表达逐渐升高,且尿液TNF-α、IL-6的活性水平逐渐升高(P0.05)。肾组织WT-1表达与肾组织IL-6、TNF-α表达及尿液IL-6、TNF-α活性水平均呈负相关(r=-0.603、r=-0.564、r=-0.563、r=-0.544,P均0.05),与尿渗透压呈正相关(r=0.821,P0.05),且与肾小管间质损害程度分级呈正相关(r=0.793,P0.01)。结论 IgA肾病系膜细胞增殖产生的细胞因子作用于足细胞并诱导足细胞凋亡、加重肾小管间质损伤。     

血清HBsAg阳性IgA肾病肾组织HBV抗原蛋白及血清和肾组织HBV-DNA检测分析

目的探讨乙型肝炎病毒（HBV）感染与IgA肾病发病的关系。方法32例肾活检冰冻切片组织HBsAg和HBcAg蛋白和42例HBsAg阳性的肾活检石蜡切片组织及其部分血清HBV-DNA的检测。结果HBsAg和HBcAg在IgA肾病肾活检组织的总阳性率为59．1％，在非IgA肾病中的总阳性率为63．6％，二者差异无统计学意义。42例肾活检组织中，仅发现有5例（11．9％）在肾活检组织中有HBV-DNA的存在。且5例均为大三阳患者，其病理诊断为系膜增生性肾小球肾炎2例，轻微肾小球病变1例，基底膜病变1例，IgA肾病仅1例。血清HBsAg阳性的患者，同时进行了42例肾活检组织的血清HBV-DNA检测，其中大三阳患者为12例，其血清HBV-DNA均为阳性，而这12例血清阳性的肾活检组织中仅有5例HBV-DNA为阳性，其余30例血清及肾活检组织中HBV-DNA为阴性。结论HBsAg和HBcAg蛋白在IgA肾病肾活检组织和非IgA肾病肾活检组织表达差异无统计学意义，表明HBV感染与IgA肾病并无直接关系。     

P-选择素与树突状细胞在IgA肾病肾小管间质病变中变化的临床意义

IgA肾病病理改变中广泛存在或伴随着肾小管间质损害 ,黏附分子介导的炎细胞浸润是肾小管间质免疫炎症反应重要前提。树突状细胞 (DC)是功能最强的专职抗原递呈细胞 ,其炎症组织迁移依赖于黏附分子P 、E 选择素介导。为此我们探讨了IgA肾病患者肾组织中P 选择素表达和DC分布变化 ,以及与肾小管间质病变之间的关系。选择经肾活检和临床资料确诊的4 5例IgA肾病患者 ,其中男 2 5例 ,女 2 0例 ,平均年龄 (4 5 .4± 1.5 )岁。根据肾小管间质病变程度分为 3组 :轻度组 2 9例 ,中度组 10例 ,重度组 6例 ;10例正常人肾组织为对照。用免疫组化法…     

IgA肾病肾组织内乙型肝炎病毒感染的发病机制研究

目的：探讨乙型肝炎病毒感染致IgA肾病肾损伤的发病机制。方法： 随机选取48例IgA肾病肾穿刺组织，参照Meadow病变分级标准分为Ⅰ-Ⅴ级5个实验组，应用Envision免疫组织化学方法检测各级肾组织内HBsAg和HBcAg；同时用直接IS-PCR技术检测其中18例IgA肾病肾组织内HBV DNA。结果： 48例IgA肾病肾组织内HBcAg和HBsAg总的阳性检出率分别为75.00%(36/48)和43.75%(21/48)；18例IgA肾病肾组织内HBV DNA阳性检出率为61.11%(11/18)；3者均表现为肾小管阳性检出率高于肾小球(P＜0.05)，但各级之间，HBcAg、HBsAg和HBV DNA检出率均无显著差异(P＞0.05)。结论： HBV参与了IgA肾病的发生，其导致肾组织损伤的机制可能主要是由细胞免疫或一系列细胞因子介导，并非病毒直接所致；肾小管上皮细胞可能是HBV感染的靶对象。     

细胞凋亡及其相关基因蛋白在阿霉素肾病中的作用

目的：探讨细胞凋亡及其相关基因蛋白在阿霉素（ADR）肾病发病中的作用及机制。 方法：复制ADR肾病模型，用超氧化物歧化酶（SOD）干预，用原位末端标记法、免疫组化法、原位杂交法分别检测细胞凋亡、基因蛋白表达、野生型p53 mRNA，同时测定肾皮质匀浆丙二醛（MDA）及谷胱甘肽过氧化物酶（GSH-Px）水平。结果：模型组皮质部肾小球、近曲及远曲肾小管细胞凋亡数和c-Myc蛋白及野生型p53转录水平、近曲和远曲肾小管细胞Bax蛋白表达、肾皮质MDA水平明显高于对照组（P<0.01），肾皮质GSH-Px活性、近曲和远曲肾小管细胞Bcl-2蛋白表达明显低于对照组（P<0.01）；SOD组肾小球和近曲、远曲肾小管细胞凋亡数明显少于模型组（P<0.01）。结论：ADR肾病病鼠的发病与肾小球、近曲和远曲肾小管细胞凋亡有关，氧自由基（OFR）可能通过上调诱导细胞凋亡的基因蛋白表达，下调抑制细胞凋亡的基因蛋白表达而引起细胞凋亡。     

实验性急性肾功能衰竭家兔肾组织学损伤与血液流变学的关系

目的 探讨实验性急性肾功能衰竭（ARF）兔肾组织形态学改变与血液流变学的关系。方法 用50%的甘油10ml/kg行兔后肢肌肉加压注射，建立ARF动物模型，观察不同时相血液流变学，肾组织形态学以及肾功能改变，结果 ARF组肾组织出现严重损害，主要为：肾小球轻度出血；肾小管广泛颗粒变性，部分肾小管上皮细胞坏死，管腔闭塞；大量管型形成；肾小管轻度出血；肾间质水肿，炎性细胞浸润；肾血管瘀血，肾功能明显下降，血液流变学指标明显异常。结论 1、甘油所致ARF兔血液流变特性明显异常；肾组织出现严重损害，2、ARF时血液流变特性恶化可能是造成肾组织形态学损害的直接原因之一。     

真武汤对阿霉素所致大鼠肾损伤的治疗作用

 目的：研究真武汤对阿霉素肾病模型（adriamycin nephropathy,AN）大鼠足细胞裂孔隔膜蛋白分子（podocin和nephrin）表达的影响,探讨其防治阿霉素肾病大鼠蛋白尿的机制。方法：采用常规生化、病理方法（包括HE染色、Masson染色及电镜）观察真武汤对AN模型所致纤维化大鼠肾功能、肾组织形态学变化及羟脯氨酸（Hyp）含量的改善作用;采用蛋白免疫印迹等方法探索真武汤对足细胞标志蛋白podocin和nephrin信号分子表达的影响。结果：模型组大鼠尿蛋白（TP）、血尿素氮(BUN)和血清肌酐（SCr）显著增加,肌酐清除率（CCr）显著下降（P<0.05）;Hyp显著增加（P<0.05）;肾组织podocin和nephrin蛋白表达水平显著降低（P<0.05）;肾小管萎缩,基底膜增厚;足突扁平、融合、消失。肾小球集中现象明显;部分肾小管扩张,肾小管上皮细胞变性,蛋白管型明显;肾间质纤维组织增生和较多炎症细胞浸润。与模型组相比,各治疗组TP、BUN和SCr均有一定程度的下降,CCr显著提高;Hyp明显下降（P<0.05）;真武汤组肾组织podocin和nephrin蛋白表达水平显著提高（P<0.05）;肾组织病变程度轻于模型组。结论：真武汤能减少阿霉素肾病大鼠肾组织羟脯氨酸含量,改善肾功能及减轻病理损伤。 真武汤降低模型大鼠蛋白尿的作用可能与其维持足细胞podocin和nephrin的表达有关。     

IgA肾病PTEN表达及其对肾间质纤维化的影响

目的： 探讨IgA肾病（IgAN）肾组织中染色体10上缺失的磷酸酶与张力蛋白同源物基因（PTEN）表达及其对肾间质纤维化的影响。方法: 选择IgAN患者47例,并详细收集资料;10例肾脏肿瘤切除后正常远端肾组织作对照。肾小管间质病变程度用Katafuchi等标准分为4组。应用免疫组化SP法检测PTEN、转化生长因子-β1（TGF-β1）、α-平滑肌肌动蛋白（α-SMA）、Ⅲ型胶原（Col Ⅲ）以及原位杂交法检测PTEN mRNA表达。结果: ①IgAN和对照肾组织中PTEN及PTEN mRNA表达部位主要在肾小管上皮细胞胞浆中,肾小球内无或仅有极少量表达。IgAN随肾小管间质病变程度加重,PTEN与PTEN mRNA表达逐渐减少,无病变组明显高于其它3组（均P<0.05）。②IgAN肾组织PTEN与PTEN mRNA表达正相关（P<0.05）;两者分别与eGFR、尿渗透压正相关（P<0.01）,与TGF-β1、α-SMA、ColⅢ、24 h尿蛋白排泄量、硬化肾小球数以及血管积分负相关（均P<0.01）。结论: IgAN中TGF-β1可能在基因转录水平下调PTEN表达,诱导肾小管上皮转分化以及细胞外基质的沉积,从而在肾小管间质纤维化过程中起到重要作用。     

IgA肾病肾组织纤连蛋白的表达

目的：探讨肾组织纤连蛋白的表达在ＩｇＡ肾病（ＩｇＡＮ）进展中的病理学及临床意义。方法：运用Ｓ－Ｐ法对６０例ＩｇＡＮ的肾活检标本进行纤连蛋白（ＦＮ）、增殖细胞核抗原（ＰＣＮＡ）的免疫组化染色，并与肾小球及肾小管间质损伤程度相对照，进行统计学处理。结果：肾小球系膜区ＦＮ与系膜细胞增生、肾小球损伤程度及肾小管间质损伤程度均呈正相关。基膜ＦＮ的出现与基膜内皮下沉着物、肾小球损伤程度及肾小管间质损伤程度均密切相关。结论：肾小球系膜区ＦＮ可作为肾活检组织病理中肾小球系膜的标志，尤其是基膜ＦＮ的聚集增多，是判断ＩｇＡＮ进展和预后不良的可靠指标     

1645例肾活检资料的流行病学特点及病理类型分析

目的：探讨江苏地区1645例肾活检资料的流行病学特点及病理类型。方法回顾性分析2009年1月~2013年6月经皮肾活检的1645例标本病理资料,并进行统计和总结。结果1645例肾活检穿刺成功1597例,成功率为97．1%;原发性肾小球疾病占78．56%,继发性肾小球疾病占18．71%,原发性肾小球疾病中以IgA肾病和系膜增生性肾小球肾炎比例较高,继发性肾小球疾病以狼疮性肾炎最多,其次为过敏性紫癜性肾炎和糖尿病肾病;苏南地区高血压性肾损伤和系膜增生性肾小球肾炎较苏北地区多见,而急性肾小管坏死和过敏性紫癜性肾炎则低于苏北地区。结论原发性肾小球疾病是江苏地区最常见的肾小球病,其中以IgA肾病发病率最高;继发性肾小球疾病以狼疮性肾炎多见;某些肾脏疾病发病率存在地域差异。     

The immunochemical characterization of the light chains in the mesangial IgA deposits in IgA nephropathy

The immunochemical characterization of the light chains of the mesangial immunoglobulin A (IgA) deposits were studied in 45 patients with IgA nephropathy. Kappa and lambda light chains were detected with direct immunofluorescence (IF) method, using monospecific rabbit anti-human anti-kappa and anti-lambda anti-sera. The glomeruli of 42 renal biopsies studied were strongly positive for lambda light chain, while only 25 specimens were positive for kappa light chain. Sixty-five percent of the biopsies showed a predominance of lambda light chain IF staining in the mesangial deposits. This IF pattern is unique as compared with similar studies on renal biopsies from patients with systemic lupus erythematosus, idiopathic membranous nephropathy, and normal postmortem renal tissue. The results indicate that mesangial IgA deposits in IgA nephropathy consist mainly of IgA with lambda light chains despite the fact that the normal ratio of kappa to lambda light-chain-containing immunoglobulin in human serum is two to one.     

Atubular glomeruli in patients with chronic pyelonephritis

In an animal model of chronic nephropathy a large proportion of the apparently normal glomeruli have been shown to be small and without connection to a proximal tubule. The present study examines the degree to which atubular glomeruli are also present in human renal disease. Eleven patients with chronic pyelonephritis (CP) and seven controls were investigated. The number of glomeruli connected to a normal proximal tubule was determined in serial sections and the volumes of individual glomeruli estimated with stereological methods. Only glomeruli with little or no sclerosis were investigated. The volume fractions of proximal tubules and interstitial tissue were estimated using point counting. The results showed that 50% of glomeruli in the CP group were connected to a normal proximal tubule, whereas 35% of the glomeruli were without any recognizable connection to a proximal tubule (atubular glomeruli). The remaining 15% were connected to an atrophic tubule. The mean volume of the glomeruli without a connection to a normal proximal tubule was only half that of glomeruli with a normal proximal tubule. No significant difference was found between the mean glomerular volume in the two groups, but the intraindividual variation of glomerular volumes was larger in the CP group. A significant negative correlation was found in the CP group between the percentage of glomeruli without connection to a normal proximal tubule and the volume fraction of proximal tubules. A significant positive correlation was found between the percentage of glomeruli that were not connected to a normal proximal tubule and the volume fraction of the interstitial tissue. This study shows that atubular glomeruli, which only can be identified in serial sections, constitute a large proportion of glomeruli in chronic pyelonephritis. Their existence could be a major reason for the irreversibility of nonglomerular chronic renal diseases.     

Tissue-type plasminogen activator and its inhibitor in human glomerulonephritis.

We carried out an immunohistochemical study of tissue-type plasminogen activator (PA) and urokinase-type PA, and their inhibitors, PA inhibitor-1 and PA inhibitor-2, using renal biopsy specimens obtained from 86 patients with various forms of glomerulonephritis. The controls were four normal renal tissue specimens. On immunofluorescent observation, granular staining for tissue-type PA was found to be distributed along the glomerular capillary walls. The fluorescence was weak in the normal renal tissue and occasionally intense in the tissues of patients with IgA nephritis, minimal change nephrotic syndrome, and lupus nephritis. PA inhibitor-1 was abundant in the glomerular epithelial cells and scarce in the mesangial area and glomerular capillary lumens of the normal renal tissues. This was confirmed by immunoelectron microscopy using gold staining. The fluorescence of PA inhibitor-1 was weaker in some specimens of nephritic tissues than in the normal renal tissues. Urokinase-type PA and PA inhibitor-2 were negative within the glomeruli in all the specimens. In the glomerulonephritic tissues which were fibrin deposition-positive, tissue-type PA expression in the glomeruli tended to be strong. An association between fibrin deposition and PA inhibitor-1 staining was not clear. These data suggest that expression of tissue-type PA in the glomeruli increases in association with fibrin deposition.     

Glomerulonephritis and superoxide dismutase]

To study the role of reactive oxygen species (ROS) in chronic renal disease, we studied the localization of Cu, Zn-superoxide dismutase (SOD) in glomeruli of patients with IgA nephropathy by immunohistochemistry on 37 kidney specimens consisting of 32 IgA nephropathy and normal parts of the 5 resected kidneys with renal tumors serving as controls. To evaluate the change in renal function, creatinine clearance (Ccr) was assessed at the time of biopsy and 1 year after the biopsy. In the normal kidney, Cu, Zn-SOD was localized in the tubular cells, and not in the glomeruli. In the kidney with IgA nephropathy, Cu, Zn-SOD was detected on the epithelial side of the glomerular capillary wall in addition to the tubular cells. The extent of localization of this enzyme was compared with the clinical findings at the time of biopsy. When Cu, Zn-SOD was stained strongly in the glomeruli, the histological change of the glomeruli was milder, and the renal function appeared to be more preserved; the decrease in Ccr one year after the renal biopsy was inhibited. These findings suggest that Cu, Zn-SOD has beneficial actions for renal function as anti-oxidative factors.     

Detection of polymeric IgA in glomeruli from patients with IgA nephropathy.

A study on the detection of polymeric IgA in glomeruli from renal biopsy specimens in patients with IgA nephropathy is described. Renal biopsy specimens were obtained from patients with IgA nephropathy. These specimens were stained with FITC-labelled anti-human J chain antisera and then examined with a fluorescent microscope. The J chain was observed in the glomerular mesangium by immunofluorescent staining. In parallel studies, renal biopsy specimens were treated with citrate buffer (pH 3.2) and the 'eluate' was neutralized by sodium hydroxide. The eluate was labelled with iodine-125, and the radiolabelled 'eluate' was fractionated by sucrose density-gradient ultracentrifugation. Polymerized IgA in the 'eluate' obtained from patients with IgA nephropathy was found to sediment predominantly as 9S to 11S using a sucrose density gradient analysis. Polymeric IgA in the fractions of the density gradient analysis was determined by anti-human IgA and anti-human J chain antisera. It was demonstrated that IgA and J chain were eluted from the glomeruli in some patients with IgA nephropathy. It is concluded that IgA deposited in the glomeruli is composed of dimers and/or larger polymers of circulating IgA in some patients with IgA nephropathy.     

Urinary IgA in IgA nephropathy and Henoch-Schoenlein purpura

To determine the concentrations and molecular forms of urinary IgA in IgA nephropathy and Henoch-Schoenlein purpura, we studied 29 patients with these IgA-associated renal diseases (IgAN). Control groups comprised 10 patients with other diverse renal diseases and 11 healthy volunteers. Urinary IgA and IgG concentrations were higher in IgAN than in either control group and correlated positively with the serum creatinine concentration as well as the urinary protein excretion (P<0.01). However, IgA/IgG ratios did not differ among the three groups. Polymeric IgA (p-IgA) in the urine predominated only in normals; in IgAN and patients with other renal diseases, monomeric IgA (m-IgA) occurred almost exclusively. Serum IgA concentrations were generally normal in IgAN; four patients had concentrations greater than 500 mg/dl. Although the fraction of p-IgA in serum (median, 18%) was increased above normal (5–10%) in 13 of 16 (81%) subjects, neither the concentration of IgA or IgG nor the amount of p-IgA correlated with the serum creatinine concentration. These data suggest that the molecular form and concentration of urinary IgA are not discriminating for IgAN and are independent of these characteristics of serum IgA.     

EVALUATION OF THE STAINING FINDINGS OF IMMUNOFLUORESCENCE IN UNFIXED OR FIXED RENAL BIOPSY SPECIMENS FROM PATIENTS WITH IGA NEPHROPATHY and MEMBRANOUS NEPHROPATHY

A study on the evaluation of staining findings of immunofluorescence in unfixed or fixed renal biopsy specimens is described. Renal biopsy specimens obtained from ten patients with IgA nephropathy and membranous nephropathy were embedded in gelatin or paraffin matrix. Renal biopsy specimens embedded in paraffin matrix were digested with 0.05% protease. The specimens were stained with FITC-conjugated anti-human IgA, IgG, IgM or C3 antisera at 4°C overnight. IgA, IgG or IgM were markedly observed in glomeruli using unfixed materials embedded in gelatin matrix or 10% neutral buffered formalin fixed materials embedded in paraffin matrix from patients with IgA nephropathy and membranous nephropathy. There was no significant difference in the intensity or distribution of IgA, IgG or IgM deposition among the two different conditions of immunofluorescence in patients with such diseases. Although the deposition of IgA using unfixed materials embedded in gelatin matrix was prominently coarse granular or lumpy in glomeruli from patients with IgA nephropathy, that of IgA using 10% formalin fixed materials embedded in paraffin matrix was fine granular and/or interrupted linear in glomeruli. It was suggested that the immunofluorescence in renal biopsy specimens embedded in paraffin matrix after digestion with protease is useful for the evaluation of immunoglobulins in glomeruli from patients with IgA nephropathy or membranous nephropathy. ACTA PATHOL. JPN. 35 : 315–321, 1985.     

ATUBULAR GLOMERULI AND GLOMERULAR CYSTS—A POSSIBLE PATHWAY FOR NEPHRON LOSS IN THE HUMAN KIDNEY?

Glomerular tufts were removed and scanning electron microscopy was used to study the interior of Bowman's capsule, in order to identify atubular glomeruli. Normal renal cortex was studied from six kidneys removed for tumour and six renal transplants removed for end-stage rejection. Atubular glomeruli occurred in normal renal cortex in less than 1 percent of glomeruli, but were more common in transplant nephropathy, representing up to 61 percent of glomeruli. Glomerular cysts were identified which also lacked a tubular connection. Both atubular glomeruli and glomerular cysts contained a contracted glomerular capillary tuft and in both, Bowman's capsule was lined mostly by parietal podocytes. It is suggested that atubular glomeruli may be precursors of the glomerular cysts. The glomerular tuft may produce filtrate which exits the glomerulus via the parietal podocytes on Bowman's capsule. In normal human kidney, the formation of atubular glomeruli by disconnection from the tubule may represent an alternative pathway for the gradual nephron loss that is associated with ageing. This process may be amplified in disease: disconnection from the tubule may be an important part of irreversible nephron damage in chronic allograft nephropathy.     

Rheumatoid factor and glomerulonephritis

It is presently unknown whether rheumatoid factors have a pathogenic role in the development of various types of glomerulonephritis with immune deposits. Three isotypes of rheumatoid factors (RFs), which are autoantibodies to IgG, were measured using the solid-phase fluorescence immunoassay in sera from patients with diffuse proliferative lupus nephritis (DPLN), membranous lupus nephritis (MLN), IgA nephropathy (IgAN) and idiopathic membranous nephropathy (MN). RF activity of immunoglobulins deposited in the glomeruli from these patients was also studied by examining the binding of the FITC-conjugated human IgG and Fc portion of IgG to the glomeruli of renal biopsy specimens. IgG, IgA and IgM RFs were significantly increased in sera from patients with DPLN, and the increase was significantly lower in patients with MLN, IgAN and MN. Human IgG bound to immunoglobulin on the glomeruli only in DPLN, but not in MLN, IgAN or MN. The Fc portion of IgG was demonstrated to be involved in this reaction. It was suggested that RFs and IgG may play a major role in immune deposits on the glomeruli in DPLN and may be involved in the development of DPLN; however, this is not likely in MLN, IgAN or MN.     

Mitochondrial enlargement and basement membrane thickening of renal proximal tubules, possible initiators of microalbuminuria in non-insulin-dependent diabetics (NIDDM).

To clarify the morphological changes in renal proximal tubules at the onset of diabetic nephropathy, we observed 177 biopsy samples from patients with Non-Insulin-Dependent Diabetics (NIDDM) using light and electron microscopy. Group I had no proteinuria (p.u.), group II had p.u. < or = 0.5 g/day, group III had p.u. > 0.5 g/day, group IV had serum creatine level (Cr) > 1.5 mg/dl. Twenty age-matched normal patients and 80 patients with IgA nephropathy were used as controls. In groups I and II, the following features were significantly different from those in the controls: spherical enlargement of mitochondria (MT) in proximal tubule cells, hypertrophy of proximal tubule cells and their nuclei, and thickening of both the proximal tubule basement membrane (TBM) and the glomerular basement membrane (GBM). Among the histological changes observed in group I, the thickness of the GBM and TBM indicated that the disease would lead to diabetic nephropathy. MT enlargement was positively correlated with nuclear and cytoplasmic enlargement of the proximal tubule cells in diabetic patients (p < 0.05), but was not correlated with other morphological changes or disease prognosis. Glomerular nodular lesions, glomerular sclerotic change, and cortical tubulointerstitial fibrosis became evident in groups III and IV. From the above, we concluded that MT enlargement and thickening of the TBM are possible causes of reduced active transport in the proximal tubules, causing microalbuminuria in diabetics, and initial impairment of post-tubule transport.