两种系膜增殖性肾炎大鼠模型的建立比较

目的　探讨快速建立系膜增殖性肾炎大鼠模型的方法。方法　 30只SD大鼠随机分 3组。正常对照组 ;葡萄球菌内毒素 (SEB)静脉注射 ,牛血清白蛋白 (BSA)隔日口服及免疫佐剂皮下注射组 ;在第二组基础上加作肝左叶切除组。 14周后分别作尿蛋白 ,IGA免疫荧光及病理组织检查。结果　肝左叶切除组蛋白尿出现的时间早于非肝切除组 ,但两组在 14周后蛋白尿定量无明显统计学差异 (P >0 0 5 ) ,对照组尿蛋白定性始终均为阴性。光镜检测轻至中度系膜扩张 ,伴系膜细胞少量增生 但两组比较无明显统计学意义 (P >0 0 5 ) ,肝左叶切除组IgA免疫荧光强于非肝切除组。结论　葡萄球菌肠内毒素静脉注射、BSA隔日口服、免疫增强剂或在上述基础上加肝左叶切除的方法都能诱发SD大鼠系膜增殖性肾炎模型。肝左叶切除组蛋白尿出现时间早 ;但于 14周后尿蛋白量无显著差异。     

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-1琢mRNA 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。     

Nephrin expression is increased in anti-Thy1.1-induced glomerulonephritis in rats

Nephrin is an important constituent of the glomerular filtration barrier and alteration of its expression is associated with severe proteinuria. In this study we show that injection of an anti-Thy1.1 antibody in rats not only induces a mesangioproliferative glomerulonephritis associated with increased proteinuria, but also leads to a sustained increase of nephrin mRNA and protein expression in renal glomeruli over a time period of 29 days. In contrast, podocin and CD2AP, two proteins shown to interact with nephrin in the slit diaphragm, are acutely downregulated at days 3-7 and, thereafter, recovered again to normal levels after 29 days. Interestingly, immunofluorescence staining of kidney sections at day 10 of the disease shows a highly heterogeneous pattern, in that some podocytes show complete absence of nephrin, whereas others show highly accumulated staining for nephrin compared to control sections, which in total results in an increased level of nephrin per glomerulus. In summary, our data show that in the course of mesangioproliferative glomerulonephritis in rats, an upregulation of nephrin expression occurs with a concomitant transient downregulation of podocin and CD2AP which may account for a highly dysregulated filtration barrier and increased proteinuria.     

血清Cys C、RBP和尿mALB检测对肾小球滤过功能及肾功能损伤诊断分析

目的:探讨血清胱抑素C(Cys-C)、视黄醇结合蛋白(retinol binding protein,RBP)及尿尿微量白蛋白(m Alb)检测对肾小球肾炎的滤过功能及肾功能损伤的诊断价值。方法:以我院2017年12月至2018年12月收治的42例肾小球肾炎患者作为观察组、39例健康体检合格者为对照组,比较两组血清Cys-C、RBP及m Alb水平,观察组不同预后者肾功能指标水平,观察组不同预后者血清Cys C、RBP及尿m ALB水平,两组血清Cys C、RBP及尿m ALB检出率。结果:观察组血清Cys C、RBP及尿m ALB水平均显著高于对照组[(2.73±0.72)mg/L vs(0.85±0.21)mg/L,(104.72±21.36)mg/L vs(37.69±8.91)mg/L,(39.51±4.02)mg/L vs(7.08±1.26)mg/L](P0.05);10例CRF/CKD和病情缓解或无进展24h尿蛋白水平无显著差异(P0.05),血肌酐、尿素氮显著高于病情缓解或无进展组[(105.76±12.37)μmol/L vs(92.09±11.01)μmol/L,(17.02±2.73)μmol/L vs(7.15±1.06)μmol/L](P0.05);CRF者血清Cys C、RBP及尿m ALB水平均显著高于病情缓解或无进展者[(3.91±0.56)mg/L vs (2.08±0.35)mg/L,(182.36±31.23)mg/L vs(47.08±9.35)mg/L,(258.68±70.27)mg/L vs (16.03±3.24)mg/L](P0.05);血清Cys C、RBP及尿m ALB阳性检出率分别为83.33%、76.19%、73.80%,联合检测率为92.85%,联合检出率显著高于单项指标检出率(P0.05)。结论:血清Cys C、视黄醇结合蛋白及尿m ALB三者联合检测有效率较高,可用作评估肾小球肾炎患者的滤过功能及肾功能损伤,观察患者预后。     

TGFbeta intercepts nuclear glycogen synthase kinase 3beta to inhibit PDGF-induced DNA synthesis in mesangial cells

Here, we demonstrate a mechanism of TGFbeta-mediated inhibition of PDGF-induced DNA synthesis in mesangial cells. TGFbeta significantly inhibited nuclear Akt phosphorylation without any effect on PDGF-stimulated phosphorylation of PDGFR at PI 3 kinase binding site (Tyr-751). Remarkably, TGFbeta inhibited cyclin D1 and cyclin E expression with concomitant decrease in CDK2 activity induced by PDGF. More importantly, we demonstrate that TGFbeta significantly abolished Akt-mediated serine-9 phosphorylation of glycogen synthase kinase 3beta (GSK3beta), thus prevented its inactivation. Expression of inactive GSK3betaK85R mutant increased cyclin D1 expression and DNA synthesis similar to PDGF. These results provide the first evidence that TGFbeta intercepts Akt kinase activity in the nucleus to block inactivation of GSK3beta, leading to attenuation of PDGF-induced CDK2 activity and DNA synthesis.     

Chemically modified siRNA prolonged RNA interference in renal disease

BACKGROUND: We previously demonstrated that transfection of synthetic short interfering RNAs (siRNAs) targeting against TGF-beta1 could be effective and therapeutic in silencing TGF-beta1 expression in glomerulus, thereby ameliorated the progression of matrix expansion in anti-Thy-1 model of glomerulonephritis. However, a major concern in applying RNAi to gene therapy is the prolonged existence of silencing potential in vivo. METHOD: We examined the duration of siRNA stability in kidney and muscle, and checked the tissue distribution of siRNase, eri-1. Thereafter, we tested the effect of chemically modified siRNA called siSTABLE on progressive glomerulosclerosis model. RESULTS: A single introduction of siRNA for EGFP (siEGFP) or its expression vector into kidney resulted in the reduction of masangial EGFP expression only for up to two weeks, while transfection of siEGFP into the pretibial muscle silenced EGFP expression unexpectedly for more than 90 days. These observations could be explained by the different expression of eri-1 between kidney and muscle. In addition, transfection of ERI-1-resistant siSTABLE for TGF-beta1 significantly reduced glomerular matrix deposition in progressive glomerulosclerosis model.Conclusion: Treatment with siRNA resistant to eri-1 may be effective and promising strategy for progressive renal disease.     

肾小球肾炎血清BUN、RBP的诊断价值分析

摘要 目的：探讨肾小球肾炎血清中尿素氮（BUN）、视黄醇结合蛋白（RBP）的表达及意义。方法：选择2018年1月至2021年1月大华医院和我院接诊的75例肾小球肾炎患者为本研究对象,设为试验组,另选择同期在我院体检的55例作为对照组,分析血清BUN、RBP水平变化情况,及其诊断价值。结果：试验组患者血清BUN、RBP水平（15.52±1.51 mmol/L、97.87±21.28 mg/L）显著高于对照组（5.07±0.97 mmol/L、42.17±10.25 mg/L）,差异显著（P＜0.05）;重度肾小球肾炎患者血清BUN、RBP水平（19.01±1.20 mmol/L、118.83±21.24 mg/L）均显著高于轻度（11.56±1.07 mmol/L、71.24±19.87 mg/L）、中度肾小球肾炎患者（14.89±1.12 mmol/L、95.75±22.13 mg/L）,差异显著（P＜0.05）;中度肾小球肾炎患者血清 BUN、RBP水平（14.89±1.12 mmol/L、95.75±22.13 mg/L）均显著高于轻度肾小球肾炎患者（11.56±1.07 mmol/L、71.24±19.87 mg/L）,差异显著（P＜0.05）;BUN诊断肾小球肾炎的AUC为0.866,95%CI为0.807~0.926,截断值为8.45 mmol/L;RBP诊断肾小球肾炎的AUC为0.957,95%CI为0.927~0.987,截断值为57.29 mg/L;联合检测诊断肾小球肾炎的AUC为0.991,95%CI为0.980~1.000,单独检测分别和联合检测曲线下面积比较均具有显著差异（Z=4.11、2.150,P＜0.05）;联合检测的特异度、准确度分别为93.41%、94.15%。结论：在肾小球肾炎患者中血清BUN、RBP的表达和病情严重程度之间存在着密切关系,可作为诊断肾小球肾炎的重要指标。     

缬沙坦对高血压合并肾小球肾炎患者Scr和GFR水平的影响

目的:探讨缬沙坦治疗高血压并肾小球肾炎临床疗效及对Scr(血清肌酐)、GFR(肾小球滤过率)水平变化的影响及临床意义。方法:选取我院收治的126例高血压合并肾小球肾炎患者为研究对象,随机分成A、B两组,各63例。A组给予低剂量(80mg/d)缬沙坦,B组给予高剂量(160 mg/d)缬沙坦。记录两组患者治疗前后SBP,Scr,GFR等临床指标变化情况及不良反应。结果:治疗前两组各项指标对比无明显差异(P0.05);治疗后,A,B两组SBP均较治疗前显著下降(P0.05);A组Scr呈升高趋势(P0.05),B组Scr呈轻微下降趋势(P0.05);A组、B组GFR变化不明显(P0.05)。A组不良反应率为4.76%,B组为9.52%,差异不显著(P0.05)。结论:对高血压合并肾小球肾炎患者给予高剂量缬沙坦,降压效果及耐受性好,毒副作用低,患者血清肌酐及肾小球虑过率水平无明显变化,值得临床推广。     

骨化三醇对系膜增生性肾炎大鼠TGF-β1的影响

目的：探讨骨化三醇对系膜增生性肾小球肾炎模型大鼠TGF-β1的作用。方法：应用抗大鼠胸腺细胞免疫血清（Anti-thy-mocyte serum,A TS）抗体一次性尾静脉注射复制大鼠MsPGN模型,将实验大鼠分为三组对照组（A组）、模型组（B组）、骨化三醇治疗组（c组）,分别与1、3、7、14天每组各处死6只大鼠,进行PAS染色,分析肾皮质损伤程度,采用ELISA法检测TGF-β1在血清中的变化,Envision免疫组化法检测TGF-β1在肾小球中的表达情况。结果：肾炎组大鼠血清及肾小球中TGF-β1表达明显高于对照组（p〈0.01）;骨化三醇治疗组大鼠血清及肾小球中TGF-β1的表达明显显著减轻（p〈0.01）。结论：骨化三醇可以抑制MsPGN模型大鼠中TGF-β1的表达,从而早期延缓肾小球疾病纤维化的发展。     

Alterations of podocytes in a murine model of crescentic glomerulonephritis

Recent observations suggest a central role of podocytes in crescent formation. In experimental glomerulonephritis podocytes disrupt the parietal epithelial layer and attach on its basement membrane, thus forming bridges between the tuft and Bowmans capsule, and they are a major constituent of crescents. In order to explain these findings we hypothesize that inflammation triggers motility in podocytes. In the present study we asked whether podocytes display alterations which suggest a migratory behavior in glomerulonephritis. Glomerulonephritis was induced in mice by injection of a rabbit serum against the glomerular basement membrane. The kidneys were perfusion-fixed 6 days later and examined by light and electron microscopy as well as by immunohistochemistry. In glomerulonephritis the apical cytoplasm of podocytes displayed numerous actin-containing microprotrusions. Cortactin, a protein involved in the regulation of actin polymerization, was predominantly expressed in foot processes of podocytes in control mice. It was redistributed to the cell body in glomerulonephritis. In untreated mice 1-integrin was restricted to the foot processes. In glomerulonephritis it was additionally found in the cytoplasm and in the apical cell membrane. Recycling of integrins is a crucial event in initiation of cell migration. ICAM-1 and CD44, the ligation of which induces migratory behaviors, were absent from healthy podocytes but expressed by some podocytes in glomerulonephritis. Thus, in glomerulonephritis podocytes display some characteristic features of migrating cells. This might explain their ability to break through the parietal epithelium and to become a constituent of early crescents.