应用高脂饮食联合维生素D3建立慢性系膜增殖性肾炎血管病变大鼠模型

目的 探讨应用高脂饮食建立慢性系膜增殖性肾炎血管病变模型的方法.方法 雄性Wistar大鼠行单侧肾切除后随机分为单纯肾切除组、单纯肾炎组、单纯高脂组、肾炎高脂组.单纯肾炎组、肾炎高脂组在单侧肾切除后3d尾静脉注射OX7抗体(100 mg/kg),1周后尾静脉连续注射OX7抗体(每次100 mg/kg,1次/周,共3次),单纯肾切除组和单纯高脂组在同一时间尾静脉注射PBS,注射抗体后第2天单纯高脂组、肾炎高脂组腹腔注射维生素D3(6万U/kg,1次/4周),同时给予高脂饲料.分别于第4、8、10周观察各组大鼠的一般情况、体重、血压、尿蛋白、血浆白蛋白、血脂、血钙、肾功能以及肾脏病理改变.结果 模型组(肾炎高脂组)大鼠第8周肾小球外的小动脉出现管壁增厚,管腔变小,平滑肌细胞减少,细胞排列紊乱,纤维组织增生.第10周单纯肾炎组和单纯高脂组肾小球外小动脉管壁轻度增厚,管腔变化不明显,模型组血管病变积分明显高于单纯肾炎组和单纯高脂组(P＜0.05).结论 通过对慢性抗Thy1肾炎大鼠加用高脂饲料并腹腔注射维生素D3的方法,可以成功建立慢性系膜增殖性肾炎血管病变模型.     

蝉蜕、僵蚕对大鼠系膜增生性肾炎作用的实验研究

目的:研究蝉蜕、僵蚕对大鼠系膜增生性肾小球肾炎(MsPGN)的治疗作用。方法:采用改良慢性血清病法制备的MsPGN模型,随机分为模型对照组、蝉蜕高剂量组、蝉蜕低剂量组、僵蚕高剂量组、僵蚕低剂量组,另设正常对照组。分别在用药5周、8周后检测大鼠24h尿蛋白;8周后处死大鼠,行血液生化指标检测和肾组织HE染色观察,免疫组化法检测TGF-β1的表达水平。结果:与模型对照组比较,用药5周后蝉蜕、僵蚕高剂量组显著降低大鼠24h蛋白尿(P<0.01);8周后各治疗组大鼠肾组织TGF-β1表达量,24h蛋白尿和血清胆固醇均显著下降,蝉蜕高、低剂量组血清白蛋白均有所升高,差异具有显著性意义(P<0.01)。肾组织形态学观察显示,蝉蜕、僵蚕高剂量组个别区域肾小球系膜细胞轻度增生,系膜区轻度增宽,管腔无挤压现象,较模型组明显改善。结论:蝉蜕、僵蚕均能有效降低MsPGN大鼠24h尿蛋白,改善脂质代谢,其作用机制可能与抑制TGF-β1的过度表达有关。     

螺内酯对单肾切除糖尿病大鼠肾脏保护作用研究

目的:研究螺内酯对单肾切除糖尿病SD大鼠的肾脏保护作用及机制。方法:48只雄性SD大鼠随机分为单肾切除对照组C组、单肾切除糖尿病模型组D组、单肾切除糖尿病螺内酯干预组S组(50mg/kg/day灌胃)。给药6周后观察各组大鼠生化指标改变,肾组织形态学改变,RT-PCR及Western Blot检测肾组织中HGF水平的变化。结果:给药6周后,D组与C组相比,血糖、24h尿蛋白定量、尿素氮、血肌酐和肾重/体重显著增高(P<0.05),螺内酯治疗后可降低糖尿病大鼠的24h尿蛋白定量和肾重/体重,但对其它指标无明显影响;形态学显示,D组与C组相比肾小球系膜区细胞外基质增多及系膜细胞增生,肾小球基底膜增厚,足细胞足突融合、消失;螺内酯治疗后上述病理改变均有不同程度的减轻;RT-PCT、Western Blot结果显示D组和S组HGF表达均增加(P<0.05),而D组HGF表达明显低于螺内酯治疗组。结论:螺内酯可能通过上调HGF的水平从而起到保护糖尿病大鼠肾脏作用。     

螺内酯对单肾切除糖尿病大鼠肾脏保护作用研究

目的：研究螺内酯对单肾切除糖尿病SD大鼠的肾脏保护作用及机制。方法：48只雄性SD大鼠随机分为单肾切除对照组C组、单肾切除糖尿病模型组D组、单肾切除糖尿病螺内酯干预组S组（50mg/kg/day灌胃）。给药6周后观察各组大鼠生化指标改变,肾组织形态学改变,RT-PCR及Western Blot检测肾组织中HGF水平的变化。结果：给药6周后,D组与C组相比,血糖、24h尿蛋白定量、尿素氮、血肌酐和肾重/体重显著增高（P〈0.05）,螺内酯治疗后可降低糖尿病大鼠的24h尿蛋白定量和肾重/体重,但对其它指标无明显影响;形态学显示,D组与C组相比肾小球系膜区细胞外基质增多及系膜细胞增生,肾小球基底膜增厚,足细胞足突融合、消失;螺内酯治疗后上述病理改变均有不同程度的减轻;RT-PCT、Western Blot结果显示D组和S组HGF表达均增加（P〈0.05）,而D组HGF表达明显低于螺内酯治疗组。结论：螺内酯可能通过上调HGF的水平从而起到保护糖尿病大鼠肾脏作用。     

低氧诱导因子在肾炎大鼠肾小管间质中的表达及氯沙坦的保护作用研究

目的:观察低氧诱导因子在肾炎大鼠肾小管间质中表达的变化以及氯沙坦对肾炎的保护作用。方法:雄性Wistar大鼠32只,随机分成假手术组、肾炎模型组、氯沙坦小剂量组、氯沙坦大剂量组各8只,假手术组不做肾脏切除,其他组在右肾切除后尾静脉注射标记抗体。给药组则按量分别灌胃给药,8周时处死。结果:大剂量给药组的血肌酐(Scr)、收缩压和24 h尿蛋白量较模型组显著降低,且小剂量给药组的血肌酐(Scr)和24h尿蛋白量较模型组也显著降低。小剂量组和大剂量组大鼠的肾间质面积较肾炎模型组均显著降低。HIF-1αmRNA大量表达于肾小管上皮细胞胞质和间质细胞胞质,且与模型组比较,小剂量给药组的HIF-1αmRNA显著降低;大剂量给药组较肾炎模型组HIF-1αmRNA的含量也显著降低。结论:氯沙坦可能可以通过影响低氧诱导因子(HIF-1α)的表达发挥对肾脏的保护作用。     

活性维生素D3对大鼠系膜增生性肾炎PCNA的影响

目的:研究活性维生素D3对大鼠系膜增生性肾小球肾炎的作用及机制.方法:应用抗大鼠胸腺细胞免疫血清(Anti-thymocyte serum,ATS)抗体一次性尾静脉注射复制大鼠系膜增生性肾小球肾炎(Mesangial Proliferative Glomerulonephritis,MsPGN)模型,将实验大鼠分为三组对照组(A组)、模型组(B组)、活性维生素D3治疗组(C组),分别与1、3、7、14天每组各处死6只大鼠,并采用Envision免疫组化法、RT-PCR法,观察PCNA蛋白、PCNAmRNA及中性粒细胞在肾小球中的表达情况.结果:肾炎组增殖细胞核抗原(Proliferating cell nuclear Antigen,PCNA)蛋白、PCNAmRNA及肾小球中中性粒细胞计数明显高于对照组(p＜0.01),并随着时间延长逐渐增高,到第7天达到高峰,第14天有下降的趋势.活性维生素D3治疗组PCNA蛋白、PCNAmRNA及肾小球中中性粒细胞计数的表达均较低(p＜0.01),其中第3、7、14天表达均显著减少(p＜0.05).PCNA蛋白、PCNAmRNA、中性粒细胞计数各指标之间呈均成正相关.结论:活性维生素D3可以抑制系膜细胞的增生,其作用机制可能是抑制炎症细胞浸润、PCNAmRNA及其蛋白的表达.     

缬沙坦胶囊联合肾炎康复片治疗糖尿病微量蛋白尿的研究

目的：研究缬沙坦联合肾炎康复片治疗糖尿病微量蛋白尿的疗效及安全性。方法：采用随机、对照原则将60例有微量蛋白尿期糖尿病患者分为缬沙坦对照组（A组）和缬沙坦联合肾炎康复片治疗组（B组）,每组各30例。A组给予缬沙坦160mg,每日1次口服;B组给予肾炎康复片（薄膜衣片）每次5粒,每日3次口服和缬沙坦160mg,每日1次口服。疗程均为12w。比较2组治疗前、治疗第4、8和12w尿白蛋白排泄率（UAER）及治疗前、后肾功能、24h尿蛋白定量等生化指标的变化及肌酐50％倍增率。结果：2组治疗后血压、Scr、CRP指标均低于治疗前（P〈0．05）,而血FBG、PBG、HbAlc、Alb、BUN、24h尿蛋白定量指标变化无差异（P〉0．05）;B组治疗后与对照组比较,B组有较低Scr、CRP值（P〈0．05）,而余生化指标及血压变化无差异（P〉0．05）,B组第8、12w时尿蛋白排泄率下降幅度低于A组（P〈0．05）,B组治疗中12w的肌酐50％倍增率低于A组（P〈0．05）。结论：缬沙坦胶囊联合肾炎康复片治疗糖尿病微量蛋白尿是有效安全的。     

缬沙坦胶囊联合肾炎康复片治疗糖尿病微量蛋白尿的研究

目的:研究缬沙坦联合肾炎康复片治疗糖尿病微量蛋白尿的疗效及安全性。方法:采用随机、对照原则将60例有微量蛋白尿期糖尿病患者分为缬沙坦对照组(A组)和缬沙坦联合肾炎康复片治疗组(B组),每组各30例。A组给予缬沙坦160mg,每日1次口服;B组给予肾炎康复片(薄膜衣片)每次5粒,每日3次口服和缬沙坦160mg,每日1次口服,疗程均为12w。比较2组治疗前、治疗第4、8和12 w尿白蛋白排泄率(UAER)及治疗前、后肾功能、24 h尿蛋白定量等生化指标的变化及肌酐50%倍增率。结果:2组治疗后血压、Scr、CRP指标均低于治疗前(P<0.05),而血FBG、PBG、HbAlc、Alb、BUN、24h尿蛋白定量指标变化无差异(P>0.05);B组治疗后与对照组比较,B组有较低Scr、CRP值(P<0.05),而余生化指标及血压变化无差异(P>0.05),B组第8、12 w时尿蛋白排泄率下降幅度低于A组(P<0.05),B组治疗中12 w的肌酐50%倍增率低于A组(P<0.05)。结论:缬沙坦胶囊联合肾炎康复片治疗糖尿病微量蛋白尿是有效安全的。     

黄芪甲苷联合骨髓间充质干细胞对糖尿病肾病大鼠的肾脏保护作用研究

目的:研究黄芪甲苷(Astragaloside Ⅳ, AS-Ⅳ)联合骨髓间充质干细胞(Bone marrow mesenchymal stem cells, BMSCs)对链脲佐菌素(Streptozocin, STZ)诱导的糖尿病肾病(Diabetic nephrology, DN)大鼠肾脏的保护作用。方法:8周龄正常雄性SD(Sprague Dawley, SD)随机选取8只作为正常对照组,其余大鼠给予腹腔注射60 mg/kg剂量STZ建立糖尿病模型。造模成功的大鼠随机分为模型组、BMSCs组和BMSCs+AS-Ⅳ组,每组8只。BMSCs+AS-Ⅳ组给予AS-Ⅳ口服灌胃治疗。BMSCs组与BMSCs+AS-Ⅳ组给予尾静脉注射BMSCs,正常对照组和模型组尾静脉注射相同剂量的无血清DMEM培养基。于第10周末收集大鼠24小时尿液,并留取肾脏组织,测定24 h尿白蛋白排泄,观察肾脏病理改变,并采用免疫组织化学方法检测肾组织中α-SMA、Desmin、Nephrin、NOX4的表达。结果:与模型组相比,BMSCs组与BMSCs+AS-Ⅳ组大鼠的肾脏病理损伤明显改善,肾小球系膜细胞增生,系膜基质增多;肾组织中NOX4、Desmin和α-SMA蛋白表达降低,Nephrin蛋白表达增加,24 h尿白蛋白排泄均明显减轻,且BMSCs联合AS-Ⅳ治疗组效果更显著(P0.05)。结论:AS-Ⅳ联合BMSCs治疗可缓解糖尿病肾病大鼠肾脏病理损伤,改善DN大鼠肾组织的氧化应激,改善肾脏足细胞转分化,从而减轻蛋白尿排泄,具有肾脏保护作用。     

CXCR4基因转染人脐带间充质干细胞移植治疗糖尿病肾病的实验研究

摘要 目的：探讨过表达CXCR4的人脐带间充质干细胞（human umbilical cord mesenchymal stem cell, hUC-MSCs）移植后对糖尿病肾病的治疗作用。方法：构建CXCR4的慢病毒表达载体，并建立过表达 CXCR4 的人脐带间充质干细胞（CXCR4-MSCs）。采用8周龄健康雌性SD大鼠75只，其中15只为正常对照组，60只为实验组。实验组糖尿病成模后一个月，将糖尿病实验大鼠60只随机分为4组：①移植CXCR4-MSCs组(CXCR4基因转染MSCs组)，即CXCR4组；②移植null-MSCs组(空质粒未转染CXCR4基因的MSCs组)，即null-MSCs；③移植MSCs组( MSCs组)；④PBS组(未移植任何的MSCs，单纯PBS注射，PBS组)。将CXCR4-MSCs、null-MSCs及MSCs消化离心，取含1×106个细胞悬液经尾静脉分别注入CXCR4-MSCs组、null-MSCs组及MSCs组大鼠体内，PBS组注射l mL PBS。干细胞治疗8周后，处死五组大鼠。各组大鼠处死前放代谢笼留取24 h尿，计算尿量，保存送检。处死前尾静脉采血检测血糖、称体重并记录。观察血糖、肾脏肥大指数、肾重、体重、24小时尿蛋白排泄量，并观察肾脏组织病理学改变。结果：60只SD雌性大鼠糖尿病模型成功率达100％，至实验8周糖尿病大鼠总共死亡14只，存活率达76.67％。实验开始后的8周，所有CXCR4组、Null-MSCs组、MSCs组、PBS组大鼠与正常组比较，体重均明显减轻(P<0.01)，血糖明显升高(P<0.01)。MSCs治疗后8周，除正常组外，其余各组大鼠血糖、肾重、肾重/体重比、24小时尿蛋白均显著增高，体重显著降低(P<0.05)；与PBS组相比，CXCR4组、null-MSCs组，MSCs组大鼠的肾重、肾重/体重比、24小时尿蛋白均明显降低(P<0.05)，体重无明显增加，血糖无明显降低(P>0.05)。CXCR4组大鼠的肾重、肾重/体重比、24小时尿蛋白较除正常组外的各组均明显降低(P<0.05)。糖尿病成模后，给予大鼠尾静脉注射干细胞悬液或等量培养液，注射后8周，除正常组外，其余各组PAS染色可见大鼠肾小球肥大，肾小球基底膜增厚、系膜增生、系膜基质增多，部分肾小球出现明显硬化，符合糖尿病肾病中期病理表现。CXCR4组大鼠肾小球系膜基质增生较其余各组大鼠减少(P<0.05)。结论：转染CXCR4的MSCs可改善糖尿病肾病。     

糖尿病肾病合并非糖尿病肾病的临床病理分析

目的：分析糖尿病肾病合并非糖尿病肾病的临床病理特点。方法：选取我院肾内科收治的临床诊断为糖尿病肾病的患者56 例,肾脏穿刺进行肾脏活体组织检查,通过病理诊断将患者分为两组,分别为糖尿病肾病组和糖尿病肾病合并非糖尿病肾病组, 比较两组患者的糖尿病病程、糖化血红蛋白、血压、血肌酐、血尿素氮、血尿酸、血清白蛋白、尿蛋白定量、血尿、视网膜病变。结果： 经肾脏组织活检,56例患者中NDRD 患者24 例(42.9%),DN患者32 例(57.1%);对24 例NDRD患者进行病理类型分类,其中IgA 肾病33.0%,膜性肾病25.0%、系膜增生性肾小球肾炎20.2%、高血压肾损害8.3%、微小病变4.2%、局灶节段硬化性肾炎4.2%、新 月体性肾小球肾炎4.2%。与DN组比较,NDRD 组糖尿病病程、糖化血红蛋白、血尿、视网膜病变均有差异(P＜0.05);而血肌酐、血 尿素氮、血尿酸、血清白蛋白、尿蛋白定量均无明显差异(P＞0.05)。结论：临床诊断的糖尿病肾病患者中有很大一部分实际上为糖 尿病肾病合并非糖尿病肾病,且以IgA 型肾病比较多见,糖尿病病程、糖化血红蛋白、血尿、视网膜病变对鉴别二者具有一定的指 导意义。     

The long pentraxin PTX3 is synthesized in IgA glomerulonephritis and activates mesangial cells

The long pentraxin PTX3 has been recently involved in amplification of the inflammatory reactions and regulation of innate immunity. In the present study we evaluated the expression and role of PTX3 in glomerular inflammation. PTX3 expression was investigated in the IgA, type I membranoproliferative, and diffuse proliferative lupus glomerulonephritis, which are characterized by inflammatory and proliferative lesions mainly driven by resident mesangial cells, and in the membranous glomerulonephritis and the focal segmental glomerular sclerosis, where signs of glomerular inflammation are usually absent. We found an intense staining for PTX3 in the expanded mesangial areas of renal biopsies obtained from patients with IgA glomerulonephritis. The pattern of staining was on glomerular mesangial and endothelial cells. Scattered PTX3-positive cells were also detected in glomeruli of type I membranoproliferative glomerulonephritis. The concomitant expression of CD14 suggests an inflammatory origin of these cells. Normal renal tissue and biopsies from patients with the other glomerular nephropathies studied were mainly negative for PTX3 expression in glomeruli. However, PTX3-positive cells were detected in the interstitium of nephropathies showing inflammatory interstitial injury. In vitro, cultured human mesangial cells synthesized PTX3 when stimulated with TNF-alpha and IgA and exhibited specific binding for recombinant PTX3. Moreover, stimulation with exogenous PTX3 promoted mesangial cell contraction and synthesis of the proinflammatory lipid mediator platelet-activating factor. In conclusion, we provide the first evidence that mesangial cells may both produce and be a target for PTX3. The detection of this long pentraxin in the renal tissue of patients with glomerulonephritis suggests its potential role in the modulation of glomerular and tubular injury.     

Ultrastructural alterations in glomerulopathy associated with hydatidosis in sheep.

In the present study we investigated the ultrastructural alterations occurring in the renal glomeruli of sheep with hydatidosis. Renal samples from 39 sheep, 34 with hydatidosis and 5 without parasitosis, were examined by transmission electron microscopy. Additionally, biochemical analysis was performed by determining serum concentrations of creatinine, urea, total protein and albumin. The ultrastructural alterations identified were the presence of dense mesangial, subendothelial and intra-membranous deposits, mesangial cell proliferation with areas showing segmental sclerosis and interposition of mesangial cells with the formation of a neomembrane. Biochemical analysis revealed a significant increase in total serum protein in the experimental group compared with the control. Our results demonstrated that glomerulonephritis associated with hydatidosis in sheep can be classified into four categories: minimal lesions, mesangial glomerulonephritis, segmental and focal glomerulonephritis and membranoproliferative glomerulonephritis, being membranoproliferative and mesangial glomerulonephritis the most predominant categories.     

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A 20-year-old man with a 10-year history of glomerulonephritis presented with a purpuric rash on his legs. A renal biopsy specimen obtained when he was 11 years old had shown mesangial glomerulonephritis; staining 9 years later for IgA had negative results. A second renal biopsy, performed when the rash was present, revealed mesangial glomerulonephritis and mesangial deposits of IgA; biopsies of the involved skin showed leukocytoclastic vasculitis. In this case isolated glomerulonephritis appeared to change to a multisystem illness, with a different immunologic character, through one of several possible pathogenetic mechanisms.     

慢性抗Thy1抗体肾炎模型的研究进展

慢性抗thy1抗体肾炎模型由于其疾病过程与人类IgA肾病及其他的系膜增生性肾炎的疾病进程相似,可以用作研究慢性肾脏病的发病机制以及新的治疗策略,本文就慢性抗thy1抗体肾炎模型的建立方法、病理演变过程、发病机制及治疗进展做一综述.     

Detection of urinary podocytes and nephrin as markers for children with glomerular diseases

The purpose of this study was to detect the urinary podocytes and its related protein, nephrin, in the urine of the children with glomerular disease in order to analyze the relationship of the clinical testing with the significance of the glomerular disease. A total of 65 children with nephrotic syndrome were selected for this study. The podocytes and nephrin were detected in the urinary sediment by indirect immunofluorescence, enzyme-linked immunosorbent assay, and Western blotting. The urinary podocytes and nephrin positive rates were 53.8% and 50.8%, respectively, in the children with glomerular disease. The serum total protein and albumin decreased in the podocyte-positive children, while the urine total protein at 24 h, urinary albumin/creatinine ratio, blood urea nitrogen, and serum creatinine were significantly elevated as compared to those of the podocyte-negative patients. Furthermore, the results were the same in the patients with positive nephrin as compared to that of the patients with negative nephrin. The podocyte number and nephrin level were significantly higher in the lupus nephritis group as compared to those of the other groups. Likewise, the podocyte number and nephrin level dramatically increased in the focal segmental glomerulosclerosis group as compared to those of the mesangial proliferative glomerulonephritis and minimal change disease groups. In addition, the podocyte numbers and nephrin expression were significantly higher in severe proteinuria group as compared to those of the mild proteinuria group. The urinary nephrin expression was positively related to podocyte and urinary albumin/creatinine ratio. We concluded that the detection of the urinary podocytes and nephrin could be taken as markers for children with glomerular disease, reflecting the type of the disease. Therefore, this can be used as a noninvasive method to evaluate the severity of the kidney disease in children.     

Downregulated expression in high IgA (HIGA) mice and the renal protective role of meprinbeta

This study discusses the critical role of the metalloproteinase meprinbeta in the progression of glomerulonephritis. Using a microarray technique, the gene expression profiles in glomeruli isolated from high serum IgA (HIGA) mice with a purity of 97% or greater were examined. HIGA mice are a valid model of human IgA nephropathy (IgAN), with the typical pathological features of this condition, including a consistently high serum IgA level as well as dominant mesangial IgA deposition and mesangial enlargement. Among the many upregulated/downregulated genes after the development of IgAN, the downregulation of meprinbeta was intriguing. The expression level of the meprinbeta gene at 40 weeks of age was 52% of that observed at 8 weeks of age (prior to the development of IgAN), although in the control BALB/c mice, a 2.19-fold elevation was seen. These results were also confirmed by semi-quantitative RT-PCR and immunostaining analyses. As meprinbeta is a subunit of metalloproteinase meprins (meprin A, meprin B) and meprins are capable of proteolytically degrading extracellular matrix (ECM) components and proteolytically processing bioactive peptides, the downregulation of meprinbeta may contribute to the progression of glomerulonephritis and the eventual glomerular scarring. This working hypothesis was examined using an in vivo meprinbeta inhibition study. The inhibition of meprins by actinonin exacerbated some parameters of renal injury in mice afflicted with anti-glomerular basement membrane (anti-GBM) antibody-associated nephritis. These in vitro and in vivo results suggest that meprinbeta may play a protective role against the progression of renal injury through the degradation of ECM and bioactive peptides.     

Molecular basis of IgA nephropathy

IgA nephropathy (IgAN) is the most common glomerulonephritis worldwide and remains an important cause of end-stage renal failure. However, the basic molecular mechanism(s) underlying abnormal IgA synthesis, selective mesangial deposition with ensuing mesangial cell proliferation and extracellular matrix expansion remains poorly understood. Notably, the severity of tubulointerstitial lesions better predicts the renal progression than the degree of glomerular lesions. The task of elucidating the molecular basis of IgAN is made especially challenging by the fact that both environmental and genetic components likely contribute to the development and progression of IgAN. This review will summarize the earlier works on the structure of the IgA molecule, mechanisms of mesangial IgA deposition and pathophysiologic effects of IgA on mesangial cells following mesangial deposition. Recently, a series of important advances in the area of communication between the glomerular mesangium and renal tubular cells have emerged. These novel findings regarding the molecular pathogenesis of IgAN will be helpful in designing future directions for therapy.