糖皮质激素抑制实验性肾炎大鼠肾组织中核因子-κB活化及单核细胞趋化蛋白-1表达

目的：探讨核因子-κB(NF-κB)活化在肾小球肾炎发病机制中的作用及糖皮质激素对NF-κB活化的调节作用。方法：肾毒血清肾炎应用兔抗鼠肾小球基底膜肾毒血清制备。模型组：注射肾毒血清后不加任何治疗,第14 d处死;糖皮质激素干预组(治疗组)：于肾毒血清注射后第1～14 d给予地塞米松每日0.125 mg/kg腹腔注射。采用免疫组化及医学图像分析系统观察肾小球及肾小管中NF-κB活化和单核细胞趋化蛋白-1(MCP-1)的表达,并分析其与蛋白尿和肾小球细胞数之间的关系。结果：模型组肾小球及肾小管中NF-κB活化较正常对照组显著上调,分别为(38.27±8.83)% vs (1.82±0.68)%和(68.46±12.94)% vs (16.89±4.47)%,P均<0.01;模型组肾小球及肾小管中MCP-1表达分别为(24.37±7.06)个/gcs和(54.78±11.49)%,较正常对照组显著升高(P<0.01),肾小球中NF-κB活化和MCP-1表达与单核细胞浸润和蛋白尿密切相关;糖皮质激素干预组肾小球及肾小管中NF-κB活化和MCP-1表达均显著下调。结论：NF-κB活化在肾小球肾炎发病机制中发挥重要作用,抑制NF-κB活化可能是糖皮质激素抗肾炎作用的机制之一。     

核转录因子-κB反义核酸对肾小球硬化细胞炎性反应的干预

目的检测大鼠肾组织核转录因子-κB（NF-κB）,血清IL-6、TNF-α的表达,探讨NF-κB与多柔比星肾病大鼠肾小球细胞炎性反应的关系及反义核酸的干预作用。方法SD雄性大鼠30只分为5组,A组（假手术组）,B组（硬化组）,C组（正义核酸组）,D组（无义核酸组）,E组（反义核酸组）,每组6只。采用右肾切除加尾静脉注射多柔比星法作肾小球硬化动物模型。第8周各组大鼠予不同药物干预,给药3 d及7 d,分别检测各组大鼠24 h尿蛋白,HE染色观察其肾组织形态学变化,利用图像分析系统计算肾小球硬化指数（GI）,应用免疫组织化学、酶联免疫吸附法检测其肾组织NF-κB p65,血清IL-6、TNF-α的表达。采用SPSS 10.0软件进行统计学分析。结果在不同的时间点,E组尿蛋白、GI、NF-κB p65、IL-6、TNF-α的表达与B组比较均显著降低,在7 d时间点,反义核酸的干预效应较3 d时间点强。结论NF-κB与肾小球硬化、细胞炎性反应存在密切关系。NF-κB反义核酸通过肾动脉给药可直接抑制肾组织NF-κB的翻译、表达,抑制肾小球系膜细胞的炎性反应,从而延缓肾小球硬化进程。     

肾病患儿尿中亚硝酸盐排泄水平分析

NO是由L-精氨酸在NO合成酶作用下产生的,具有很强的反应活性的自由基气体,参与了神经传导、血压的调控。在肾脏NO可调节入球动脉张力及肾小球系膜细胞的增殖,近期研究还表明NO参与了大鼠肾炎模型的损伤过程,如由抗胸腺细胞抗体诱导的大鼠系膜增殖性肾炎,抑制NO合成酶可减少尿蛋白排泄量及系膜基质的扩张等。NO性质极为活泼,体内可迅速分解为NO_2~-/NO_3~-,后者性质稳定,可代表体内NO代谢情况,本文以镉柱比色     

儿童系膜增殖性肾小球肾炎α-平滑肌肌动蛋白表达的观察及临床意义

本文应用免疫组化技术及真彩医学图像分析系统观察 32例系膜增殖性肾小球肾炎患儿α 平滑肌肌动蛋白 (α ＳＭＡ)的表达 ,以期为抑制系膜细胞的过度增殖及肾小球疾病的治疗提供理论依据。病例选择 :选择 1997～ 1999年南京医科大学儿肾研究中心经肾穿刺活组织检查诊断的系膜增殖性肾小球肾炎 32例 ,男 2 2例、女 10例 ;年龄 3～ 12岁 ,平均 (8 1± 3 2 5 )岁 ;临床诊断为肾病综合征 2 1例 ,单纯性血尿 6例 ,单纯性蛋白尿 5例 ;病程 0 5个月至 5年。其中轻度系膜增殖 13例、中度 9例、重度 10例。临床诊断标准、治疗方案及疗效判断标准按“…     

罗格列酮对大鼠系膜增生性肾炎的治疗作用

目的 研究罗格列酮对大鼠系膜增生性肾炎(MsPGN)的治疗作用并探讨其可能的作用机制,寻找治疗系膜增生性肾炎的新途径.方法 大鼠随机分为3组:李白对照组(A组)、模型对照组(B组)、罗格列酮治疗组(C组).B、C组以四氯化碳及牛血清白蛋白灌胃制备系膜增生性肾炎模型,制模成功后.A、B组予生理盐水,治疗组予罗格列酮分别灌胃均8周,每周测各组大鼠24 h尿蛋白排泄量及尿红细胞计数.实验结束时,测血清白蛋白、血糖、肝肾功能、肾脏系膜细胞及系膜基质增生情况(光镜、PAS染色)及各组大鼠肾脏转化生长因子-β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)表达水平.结果 治疗组大鼠24 h尿蛋白排泄量、尿红细胞汁数、TGF-β1及α-SMA表达水平、肾小球PAS染色灰度值较模型组均明显减少,血清白蛋白明显升高,差异均有统计学意义.3组之间血精及肝肾功能比较差异无统计学意义.结论 罗格列酮对大鼠系膜增生性肾炎具有治疗作用,其作用机制之一可能是通过抑制TGF-β1及其下游因子表达.     

系膜细胞来源的白细胞介素-13对系膜细胞细胞因子基因表达的研究(英文)

目的　白细胞介素 1 3(IL 1 3)是新近发现的一种抗炎性细胞因子 ,其在肾小球肾炎中的作用尚不清楚 ,该研究探讨脂多糖 (LPS)对体外培养的人肾小球系膜细胞 (HMC)表达IL 1 3作用以及IL 1 3对HMC促炎性细胞因子、趋化因子和促纤维化因子基因表达的影响。方法　体外培养HMC ,加入不同浓度的LPS和 (或 )IL 1 3后 ,用逆转录 -聚合酶链反应和ELISA检测HMCIL 1 3mRNA表达和细胞培养上清液中IL 1 3蛋白含量 ;应用核酸酶保护法检测HMC肿瘤坏死因子 α(TNF α)、白介素 - 1α(IL 1α)、白介素 - 1 β(IL 1 β)、单核细胞趋化蛋白 1(MCP 1 )、白介素 8(IL 8)、转化生长因子 - β1 (TGF β1 )mRNA的表达。 结果　未予LPS刺激的HMC不表达IL 1 3mRNA和蛋白 ;LPS呈剂量依赖性和时间依赖性诱导HMC表达IL 1 3mRNA和分泌IL 1 3蛋白。HMC受LPS刺激后 1 2h即可表达IL 1 3mRNA ,4 8h达高峰 ,72h仍维持在较高的水平。HMC受LPS刺激后 2 4h ,其培养上清液中检测到IL 1 3蛋白 ,4 8h和 72h进一步增加。外源性IL 1 3呈剂量依赖性地抑制LPS诱导的系膜细胞TNF α ,IL 1α ,IL 1 β ,MCP 1 ,IL 8,TGF β1mRNA的表达。应用抗IL 1 3抗体中和内源性IL 1 3后 ,上述炎症因子表达增强。结论　IL 1 3是HMC自分泌因子。IL 1 3可抑制LPS诱导     

增生性肾炎患者系膜细胞增生与细胞周期调控蛋白的相关研究

目的 观察增生性肾炎患者肾小球系膜细胞（MC）在不同增生程度时细胞周期调控蛋白表达水平的变化。方法 采用免疫组织化学技术,检测34例肾不小球肾炎患者肾刺标本中细胞正性调控蛋白周期素D1（cyclinD1)、负性调控蛋白P21^CIP1(P21)、P27^KIP1(P27)和增殖细胞核抗原（PCNA）表达。结果 在正常对照及非增生性肾炎组中,cyclin D1、P21不表达,P27呈高表达。在增生性肾炎组中,cyclin D1、P21和P27阳性表达主要在系膜区,随着系膜增生程度的加重,肾小球cyclinD1、P21表达增加,并与PCNA呈正相关（P均＜0．01）;P27则表达减少并与PCNA呈负相关（P＜0．05）;肾小球P27表达还与血肌酐之间呈显著性负相关（P＜0．01）。结论 在人类增生性肾炎中,cyclin D1、P21和P27参与MC的增生;P27在肾组织的表达量可能与疾病进展密切相关。     

小儿肾小球疾病肾活检:附29例次资料分析

本文报道对27例肾小球疾病患儿行29次经皮肾活检,成功28次。其中肾病综合征14例,其它肾小球疾病13例。病理结果系膜增殖肾炎14例,微小病变4例,IgA肾病4例,膜性肾病、膜增殖性肾炎、弥漫增殖性肾炎、新月体肾炎及弥漫肾小球硬化各1例。肾活检对明确肾病的病理类型、确诊肾脏疾病、指导治疗、判断预后有较大意义。小儿肾活检只要准备充分、操作熟练是安全的。本组除4例肉眼血尿外,无其它严重并发症。本文还提出具体操作注意点。     

黄芪对实验性IgA肾病大鼠肾小管间质损害及NF-κB MCP-1表达的影响

目的：观察黄芪对实验性IgA肾病大鼠肾间质损害及肾组织核转录因子-κB （NF-κB）、单核细胞趋化蛋白（MCP-1）表达的影响,探讨黄芪防治IgA肾病肾小管间质损害的作用机制。方法：28只SD大鼠分为模型组、干预组、对照组3组。模型组和干预组复制IgA肾病模型,干预组给予黄芪颗粒剂口服,并以正常SD大鼠为对照组。应用全自动生化分析仪检测尿红细胞、24 h尿蛋白定量、尿N-乙酰-β-D-氨基葡萄糖苷酶（NAG）活性;应用免疫荧光法检测肾组织冰冻切片IgA免疫复合物沉积情况,利用免疫组织化学方法,观察大鼠肾组织NF-κB p65,MCP-1表达的影响;半定量评分法计算病理积分以观察肾脏病理损害程度。结果：①干预组大鼠的尿红细胞、尿蛋白、尿NAG酶含量较模型组明显降低,差异有统计学意义(P<0.01);②模型组大鼠肾组织NF-κB、MCP-1的表达与干预组、对照组相比均明显增高(P<0.01),差异有统计学意义;③干预组大鼠肾脏病理评分较模型组明显降低,差异有统计学意义(P<0.01 )。结论：黄芪能减轻模型组大鼠的尿红细胞数、尿蛋白和尿NAG酶活性;能减轻IgA肾病模型大鼠肾小管间质病理损害,其减轻肾小管间质损害作用可能与下调NF-κB,MCP-1表达有关。     

吗替麦考酚酯治疗以蛋白尿为主的儿童紫癜性肾炎疗效观察

目的 评价吗替麦考酚酯(MMF)分散片联合小剂量激素治疗以蛋白尿为主的儿童难治性紫癜性肾炎的临床疗效及安全性.方法 选取19例以蛋白尿为主的难治性紫癜性肾炎儿童作为研究对象,其中5例在MMF治疗之前接受肾活检(轻度系膜增殖性肾炎2 例,中度系膜增殖性肾炎伴新月体形成1例,重度系膜增殖性肾炎1例,局灶节段性肾小球硬化1例).泼尼松0.5 ～ 1.0 mg/(kg·d),平均0.74 mg/(kg·d),尿蛋白转阴2周后逐渐减量,半月减5 ～ 10 mg至5 ～ 10 mg/d维持;MMF分散片20 ～ 25 mg/(kg·d),2次/d.随访1 ～ 1.7年.治疗期间定期复查血常规、尿常规、肾功能、肝功能以及24 h尿蛋白定量等,治疗3 个月后进行疗效及安全性评价.结果 MMF分散片联合小剂量激素治疗前、后24 h 尿蛋白定量和血清白蛋白差异有统计学意义(P ＜ 0.01).治疗3 个月后19 例中完全有效17 例,部分有效2 例.1例应用MMF后2个月合并水痘,1例出现了胃肠道症状,2例一过性白细胞减少.结论 吗替麦考酚酯分散片联合小剂量泼尼松能有效缓解以蛋白尿为主的儿童紫癜性肾炎尿蛋白,改善病情,且近期不良反应少,安全性高,依从性好.     

Effects of podocyte injury on glomerular development

To study the effects of podocyte injury on glomerular maturation and underlying mechanisms of such effects, puromycin aminonucleoside (PAN) was given to neonatal mice at 1 d post partum (1 dpp). Mice with PAN injection had smaller kidney weight (KW) and body weight (BW) at all times and smaller KW/BW at 4, 8, and 12 dpp versus normal saline (NS) controls. Electron microscopy (EM) revealed nearly complete podocyte foot process effacement and segmental microvillous transformation as early as 2 dpp, preceding proteinuria. PAN-injected kidneys showed significantly fewer glomerular capillary loops and decreased glomerular maturation index, as well as less CD31+ endothelium in cortical glomeruli at 12 dpp versus NS controls. Glomerular mesangial injury and glomerulosclerosis along with proteinuria were noted in PAN-injected kidneys starting from 30 dpp. Systolic blood pressure was increased significantly by 60 dpp in PAN mice. PAN mice also had significantly decreased Flk-1 and Tie2 mRNA expression and increased angiopoitein-1 (Ang-1) expression, without change in vascular endothelial growth factor (VEGF) at 2 dpp versus NS. Our study shows that podocyte injury in neonatal mice kidneys alters the expression of key capillary growth modulators in glomeruli, leading to abnormal development of glomerular capillaries, with subsequent development of proteinuria, hypertension, and glomerulosclerosis.     

Screening for renal disease in school children: experience in Japan

A program of urine screening for asymptomatic hematuria and proteinuria in school children has been conducted since 1973 by the Ministry of Education in Japan with great success in the early detection of asymptomatic renal disease. Prevalence of isolated persistent hematuria in school children was approximately 05%. Prevalence of isolated proteinuria and proteinuria with associated hematuria was 0.08% of primary school children and 0.4% of junior high school children. In order to know whether this nationwide program for 13 years causes some changes in the epidemiology of chronic glomerular disease in Japan, a multicenter survey was conducted on the number of patients with renal diseases. 70–80% of IgA and non-IgA mesangial proliferative glomerulonephritis and 65 to 80% of MPGN were detected by mass urine screening at school. Severe glomerular lesions were more frequently observed in children with chance proteinuria and hematuria, as well as IgA and non-IgA mesangial proliferative glomerulo nephritis who have severe proteinuria. Mild glomerular change was more frequent in patients with MPGN, IgA and non-IgA mesangial proliferative nephritis who were detected by our screening program rather than those seen with some of the nephritic signs and symptoms at diagnosis The above evidence appears that screening program may open the way for the early management of these diseases, especially for which treatment is already established.     

Mechanism of the unique susceptibility of deep cortical glomeruli of maturing kidneys to severe focal glomerular sclerosis

The alterations in single glomerular hemodynamics, glomerular size, and development of glomerular sclerosis after subtotal nephrectomy were assessed in approximately 32-d-old young and greater than 3-mo-old adult Munich-Wistar rats. In 6 wk, young rats developed more pronounced glomerular sclerosis and more marked elevation in blood urea nitrogen. The deep (versus superficial) cortical region of young rats was characterized by having a greater number of glomeruli with advanced sclerosis. Single nephron glomerular filtration rate of superficial glomeruli of the young increased to a much greater extent than whole kidney glomerular filtration rate, whereas there were comparable post-subtotal nephrectomy increases in whole kidney glomerular filtration rate in these two age groups, indicating that the deep glomeruli were exposed to a lesser hemodynamic load than were the superficial. Since the remnant nephrons of young and adult rats achieved equally high glomerular pressures and comparably large glomerular size shortly after subtotal nephrectomy, the unique susceptibility of young glomeruli to sclerosis is attributed to the intrinsic property of these glomeruli, rather than the abnormal hemodynamics or stimuli promoting hypertrophy and mesangial matrix accumulation imposed upon the glomeruli.     

板层蛋白在病理肾组织中的分布及其mRNA的表达

目的 研究板层蛋白在肾小球疾病时质或量的改变,探讨它们在肾脏中的生理功能和病理学意义。方法 本研究以板层蛋白α1,α2,β1,β2,γ1结构链为观察对象,用免疫组织化学和原位杂交技术研究它们在48例肾小球疾病病理肾活检组织中的表达和分布是否发生质或量的改变。结果LNα1,α2,β1和γ1的mRNA在伴系膜增生的肾小球内表达有不同程度的增加,增生的系膜细胞是主要的细胞来源。在肾小球系膜增殖节段和硬化病灶附近伴有LNα1和γ1的蛋白增多和LNα2,β1蛋白的异常表达,而LNβ2链在相应节段的系膜区和GBM表达出现不同程度的下降。结论在系膜增生的肾小球疾病中,增生的系膜细胞是肾小球内板层蛋白量的积聚和异常合成的重要细胞来源。在肾小球疾病中,板层蛋白的合成出现量和质的变化,这可能是肾小球疾病进展和恶化的物质基础。     

整合素连接激酶表达与儿童紫癜性肾炎肾小球损害的关系

目的研究表明整合素连接激酶(integrin-lin kedkinase,ILK)在肾脏疾病的发生、发展中起着重要作用。该研究旨在探讨ILK与儿童紫癜性肾炎(HSPN)肾小球损害的关系。方法188例HSPN患儿按照国际儿童肾脏病组织(ISKDC)关于HSPN肾脏病理分级分成5组,即≤Ⅱa级组、Ⅱb级组、Ⅲa级组、Ⅲb级组、≥Ⅳ级组。对照组为薄基膜肾病患儿。采用免疫组织化学方法检测对照组及HSPN患儿肾小球ILK的表达,分析肾小球ILK表达与肾小球病理损害、尿蛋白排泄量之间的关系。结果①ILK在肾小球的表达:对照、≤Ⅱa级、Ⅱb级、Ⅲa级、Ⅲb级、≥Ⅳ级组6组患儿肾小球ILK的阳性表达面积分别为:(3.35±1.01)%、(4.88±1.13)%、(9.64±1.36)%、(11.27±1.68)%、(17.42±3.0)%、(20.62±2.32)%,6组间比较,差异有非常显著性(P<0.01)。②随着患儿尿蛋白排泄量的增加,肾小球ILK表达逐渐增强,尿蛋白阴性组、尿微量白蛋白组、显著尿蛋白组及大量尿蛋白组间的ILK表达差异有非常显著性(P<0.01)。结论ILK与儿童HSPN肾小球病理损害及蛋白尿的发生密切相关。     

MICROANGIOPATHIC GLOMERULOPATHY IN CHILDREN WITH SICKLE CELL ANEMIA

We studied kidney biopsy specimens from three children with sickle cell anemia and microangiopathic glomerulopathy. One child also had cyanotic congenital heart disease. Laboratory evaluation revealed proteinuria and normal serum creatinine in all and normal serum complement in two of the three children at the time of biopsy. In all biopsies, glomeruli were enlarged with diffuse hypercellularity and focal segmental mesangial interposition; capillary loop lumens were congested with sickled erythrocytes. Immune labeling identified segmental immunoglobulin G, C3, and properdin over the glomerular capillary loop walls in each case. Ultrastructurally, the subendothelial zone of the glomerular basement membrane was widened with new lamina densa formation with focal mesangial interposition. The glomerular lesion we describe in these children may be due to endothelial injury related to the altered erythrocytes, glomerular hemodynamics, and the hypercoagulable state characteristic of sickle cell disease.     

不同病理类型儿童狼疮性肾炎肾小球细胞凋亡的变化

目的检测不同病理类型狼疮性肾炎(lupusnephritis,LN)患儿肾组织细胞凋亡与增殖,分析LN患儿肾组织细胞凋亡与增殖的关系。方法用原位末端标记法(TUNEL)检测21例Ⅳ型、6例Ⅴ型LN患儿肾组织及9例对照肾组织中的细胞凋亡,用免疫组织化学法检测其细胞增殖(PCNA,增殖细胞核抗原),同时用免疫组织化学及图像分析法检测细胞凋亡相关基因蛋白PDCD5、Caspase-3的表达。结果1.Ⅳ型LN患儿肾小球凋亡细胞数(0.93±0.28)、增殖细胞数(9.97±1.90)及P/A值(11.15±2.49)均明显高于Ⅴ型LN患儿相应指标(0.66±0.11),(4.82±1.46)和(7.25±1.71),P<0.05,P<0.01和P<0.01。2.Ⅳ型和Ⅴ型LN患儿肾组织PDCD5、Caspase-3表达:PDCD5在Ⅳ型LN患儿肾小球的表达强度(0.08±0.02)与V型LN患儿肾小球的表达强度(0.07±0.03)无差异,P均>0.05;Caspase-3在Ⅳ型LN患儿肾小球的表达强度(0.25±0.05)明显高于Ⅴ型LN患儿肾小球的表达强度(0.16±0.03),P<0.01。结论Ⅳ型LN患儿相对于细胞增殖的程度而上调的细胞凋亡远不如Ⅴ型LN患儿;Caspase-3参与LN患儿肾小球细胞凋亡,而PDCD5未起主要作用或其促凋亡作用被抑制。     

Role of bone marrow cells in the healing process of mouse experimental glomerulonephritis

Recent studies have shown bone marrow (BM) cells to differentiate into a variety of cell types and to thereby participate in the reconstitution of damaged organs. In the present study, we examined the extent to which BM-derived cells are incorporated into glomeruli during recovery from experimentally induced nephritis. To investigate the localization of BM cells in glomeruli, chimeric mice were prepared by transplanting BM cells from green fluorescent protein (GFP) transgenic mice into wild-type mice. Five weeks later, glomerulonephritis was induced by intravenous injection of Habu snake venom. Groups of mice were then killed every few days for 42 d, and harvested kidney samples were subjected to immunohistochemical and immunoelectron microscopic analyses with the aim of detecting the presence of GFP(+) cells within glomeruli. Chimeric animals injected with Habu venom developed proliferative glomerulonephritis within 1-3 d. The lesion gradually subsided and the glomerular structure returned to normal within 42 d. Consistent with the disease course, large numbers of GFP(+) cells were present within glomeruli on d 1-3, but most had disappeared by d 7. Nevertheless, some GFP(+) cells did remain within glomeruli showing mesangial proliferative changes, and were found to express thrombomodulin (TM), a specific endothelial cell marker. These GFP-TM-double-positive cells accounted for a mean of 1.31-2.24% of the total glomerular nuclei from d 7 through d 42, levels that remained stable for at least 12 mo. It thus appears that BM cells can give rise to endothelial cells that participate in the remodeling of glomeruli.