肝细胞生长因子 转化生长因子-β_1在IgA肾病肾组织的表达及意义

目的对IgA肾病肾组织进行肝细胞生长因子(HGF)、转化生长因子(TGF)-β1水平测定,探讨其与IgA肾病肾脏病理分型、Katafuchi评分及临床指标的关系。方法①76例IgA肾病患者肾脏组织被分为轻度系膜增生(A组)、中到重度系膜增生(B组)、局灶增生或增生硬化性IgA肾病(C组)3组;②采用Katafuchi的半定量标准评分,计算相应的肾小球总积分、系膜增生程度积分、球性硬化积分、血管积分;③检测患者的血压、尿蛋白定量(24h)、血清肌酐(Scr)、内生肌酐清除率(Ccr)、血清白蛋白(Alb)和脂蛋白a[Lp(a)];④免疫组织化学及反转录-聚合酶链反应(RT-PCR)方法测定患者肾组织切片中HGF、TGF-β1水平,收集数据进行统计分析。结果①随着病变的加重,各病理指标相应积分及血压、Lp(a)、尿蛋白定量(24h)、Scr逐渐增加,Ccr、Alb逐渐降低;②与A组比较,B组肾组织TGF-β1明显增多(P<0.01),C组显著降低(P<0.05);与A组比较,B、C组HGF面积显著降低(P<0.01);③肾组织HGF阳性面积与Ccr、Alb呈正相关(r=0.76、r=0.60),与肾组织TGF-β1阳性面积无相关,与血压、、Scr、Lp(a)、肾小管间质积分、肾小球总积分、系膜增生程度积分、球性硬化积分、血管积分呈负相关(r分别=-0.69、-0.75、-0.70、-0.67、-0.74、-0.69、-0.35、-0.39、-0.32,P<0.01)。结论①各临床病理指标与病理分级有相关性,提示动态监测临床病理指标对判断IgAN的预后有意义;②IGF-β1参与肾纤维化过程;随肾脏病变的加重HGF含量降低,可导致其纤维化作用减弱。     

Protection of rhein on IgA nephropathy mediated by inhibition of fibronectin expression in rats

Objective:

To investigate the protective effects of rhein on IgA nephropathy (IgAN) in the rat model.

Materials and Methods:

Twenty-eight female sprague dawley rats were divided randomly into four groups, namely control, IgAN, rhein-prevented and rhein-treated. The pathologic changes on renal tissue were observed by the H and E, staining and the amount of urinary red blood cells and 24-h urinary protein excretion were measured. The glomerular deposition of immune globulin A (IgA) was measured by immunofluorescence staining. Fibronectin (FN) and α-smooth muscle actin (α-SMA) expression on renal tissue were measured via immunohistochemistry.

Results:

The model of IgAN was established according to Bovine serum albumin-Lipopolysaccharide-Carbon tetrachloride protocol, which was evidenced by histological structural lesions of glomeruli, IgA deposition and urinary measurement. Histological examination of kidney sections from both rhein-prevented group and rhein-treated group showed that glomerular hypertrophy, mesangial expansion, excessive extracellular matrix, and renal capsule dilation were markedly ameliorated compared with IgAN group. Moreover, rhein treatment significantly reduced IgA deposition in glomerulus, the volume of urinary red blood cells and 24-h urinary protein excretion. More importantly, increased FN expression in IgAN was back to normal level in rhein-prevented and rhein-treated group, which was along with the reduction of α-SMA expression in renal tissues.

Conclusions:

These findings indicate that rhein prevents the development of glomerulosclerosis and halts the progression of IgAN via inhibition of FN and α-SMA expression.KEY WORDS: IgA nephropathy, rat, rhein     

IgA肾病肾血管病变与临床病理之间的关系

目的：探讨IgA肾病（IgAN）肾血管病变的临床和病理学意义。方法：将177例经肾活检确诊的原发IgAN患者分为肾血管病变组及无肾血管病变组,比较两组临床指标及病理改变与血管病变之间的关系。结果：177例IgAN患者中有肾血管病变者占65．5％。与无肾血管病变组相比,肾血管病变组血压升高、尿蛋白增多、血肌酐升高、病理分级加重,肾小球补体C3沉积更明显。结论：IgAN肾血管病变与临床和病理变化显著相关,可以作为判断预后的一项重要病理指标。     

Toll-like receptor 4 is involved in a protective effect of rhein on immunoglobulin A nephropathy

Objectives:The objective was to investigate the protective effects of rhein on renal histology change and the effects of rhein on renal tissue toll-like receptor (TLR) 4, TLR9, transforming growth factor-β1 (TGF-β1) expression in immunoglobulin A nephropathy (IgAN) rats.Results:The biochemical parameters results of IgAN model rats showed that the 24-h UP excretion and ALT concentration were much higher, and TP concentration was much lower than those of the control group (P < 0.05). Granule-like or mass-like IgA depositions in the mesangial area, glomerular hypercellularity, hyperplasia of mesangial matrix, and tubulointerstitial fibrosis were found in IgAN group. Rhein-prevented and rhein-treated both improved the biochemical parameters and relieved renal pathological injury. The expressions of renal tissue TLR4, TGF-β1, but not TLR9 were significantly elevated in IgAN model rats (P < 0.05). Rhein-prevented and rhein-treated both inhibited TLR4 and TGF-β1 expressions.Conclusion:Rhein significantly improved the serum and urine biochemical parameters, and attenuated the glomerular pathological changes and tubulointerstitial fibrosis in IgAN rats. The mechanism may involve inhibition of renal TLR4 and TGF-β1 secretion.KEY WORDS: Immunoglobulin A nephropathy, rhein, toll-like receptor, transforming growth factor-β1     

贵州省IgA肾病临床与病理特征相关性分析

目的 研究贵州省IgA肾病(IgAN)患者的临床表现与病理类型两者之间的关系,以探讨贵州省IgAN的发病率,进一步评价肾活检在肾脏病诊治中的作用及临床意义.方法 方法对730例有肾穿刺活检适应证的患者行B超介导下肾活检,并作光镜、免疫荧光、电镜检查,对其中确诊为lgA肾病者作病理及临床特点分析.结果 (1)贵州省lgA...     

Detection of immunoglobulin depositions by immunofluorescence and/or immunoperoxidase (P-AP method) in patients with IgA nephropathy

Deposition of immunoglobulins in the glomeruli by immunoperoxidase (IP) and/or immunofluorescence (IF) using unfixed materials obtained from 25 patients with IgA nephropathy (IgAN) was evaluated. The results indicated that the deposition of IgA, IgM or IgG in the glomerular capillary walls using a peroxidase anti-peroxidase (P-AP) method was more prominently detected than by using IF. The results of double staining indicated that both P-AP and IF were able to stain mesangial and capillary depositions of IgA was more widely detected by P-AP than by IF. It is concluded that the P-AP method has the advantage of indicating the deposition of immunoglobulins in glomerular capillary walls, and evaluating the histopathological significance together with IF in patients with IgAN.     

慢性肾功能不全患者肾穿刺活检的风险与价值研究

目的回顾性分析慢性肾功能不全（CRI）患者经皮肾穿刺活检（PRB）的风险与价值。方法对符合条件的50例CRI患者进行PRB,行光镜、免疫组化染色和选择性电镜检测,观察肾穿刺组织肾小球个数、病理类型、诊断以及穿刺并发症。结果肾组织标本合格率为96%。病理类型前三位为增生性肾小球硬化症21例（42%）,IgA肾病（IgAN）9例（18%）,狼疮性肾炎（LN）8例（16%）。PRB后修改诊断5例（10%）,明确病因3例（6%）,诊断修正率为16%,根据病理结果决定治疗方案16例（32%）,治疗方案修正率为26%。并发症以镜下血尿最常见,共48例（98%）,肾周小血肿11例,肾周大血肿2例,腹膜后血肿1例。结论原发性肾脏疾病肾穿后病理结果多为慢性病变,大部分患者的诊断治疗方案不受肾穿刺病理结果的影响,且肾周血肿出现机率较多且较大,偶可合并腹膜后血肿,但狼疮性肾炎表现为慢性肾衰时其肾脏病理的活动指数仍较高,少数病例肾脏病理显示新月体肾炎,仍需应用免疫抑制剂以改善预后。     

血清乙型肝炎病毒阴性肾小球肾炎与乙型肝炎病毒感染关系的探讨

目的了解乙型肝炎病毒(HBV)感染与肾小球肾炎的相关性,探讨肾组织中HBV抗原的来源及其与病理变化的关系,以有助于进一步研究HBV相关性肾炎的发病机制。方法应用免疫组织化学(SP)法检测96例血清HBV感染标志阴性,5例血清HBV感染标志阳性肾炎肾组织中的HBV抗原[HBV表面抗原(HBsAg)、HBV核心抗原(HBcAg)],用原位杂交方法检测32例肾炎组织HBV抗原阳性的(包括5例血清HBV感染标志阳性者)中的HBV-DNA。5例肾组织中检出HBV抗原的膜性肾病(其中1例血清HBV感染标志阳性)作透射电镜观察。结果发现血清HBV感染标志阴性肾炎肾组织中HBV标志物沉积与在血清HBV标志阳性肾炎中一致:HBV抗原除在肾小球沉积外,肾小管常有阳性表达,并且肾小管HBcAg阳性率高于其在肾小球中的表达(43.6%vs23.8%)。肾小管HBcAg阳性组的肾组织病变程度明显严重于肾小管HBcAg阴性组,且差异有显著性(P<0.05)。27例血清学阴性肾炎中8例肾组织中HBV-DNA阳性,主要沉积于肾小管上皮细胞质。电镜观察显示,血清HBV标志阴性膜性肾病表现为基底膜弥漫增厚,电子致密物在上皮下沉积及上皮足状突细胞融合;血清HBV标志阳性膜性肾病表现为基底膜不均匀增厚,基膜内小而散在的高密度电子致密物沉积。结论血清HBV感染与肾炎[膜性肾小球肾炎(MGN)、膜增生性肾小球肾?     

Determination of tissue type plasminogen activator in the glomeruli of patients with IgA nephropathy

The detection, by immunofluorescence, of tissue type plasminogen activator (t-PA) in the glomeruli of 50 patients with IgA nephropathy was described. Patients with t-PA deposits revealed a higher frequency in the glomerular deposition of fibrin and/or fibrinogen than did patients without t-PA deposits. There also was a distinct relationship between the level of serum t-PA and the degree of renal tissue injury in patients with glomerular t-PA deposits. It was concluded that the deposition of t-PA along with fibrin and/or fibrinogen might reduce the effectiveness of fibrinolysis in the glomeruli of patients with IgA nephropathy.     

Comparison of endothelin-A and endothelin-B receptor distribution visualized by radioligand binding versus immunocytochemical localization using subtype selective antisera

Molecular studies have predicted the existence in human tissue of splice variants and modifications to the amino acid sequence of endothelin receptors that may modulate function. Endothelin-A (ETA) receptors were visualized by ligand binding and autoradiography to the renal vasculature throughout the cortex and medulla, including the large arcuate arteries, adjacent veins and arterioles. Lower binding densities were visualized to the vasa recta and glomeruli. A similar pattern of staining was revealed by ETA selective antisera, with the higher resolution demonstrating ETA receptors confined to smooth muscle cells. Staining was also detected to the vasa recta and glomeruli. Ligand binding revealed a more heterogeneous endothelin-B (ETB) receptor distribution with high densities concentrated in the medulla. Three different site-directed ETB antisera demonstrated a similar pattern of staining to the endothelium lining all renal vessels but not to the smooth muscle. Staining was also detected to glomerular endothelial cells as well as epithelial cells lining the renal tubule, particularly the collecting ducts, consistent with high binding densities observed in the medulla by autoradiography. There was no evidence for a differential distribution in either ETA or ETB receptors visualized by the two techniques that might have indicated modified receptors or further subtypes in the human kidney.     

山茱萸提取物对慢性肾炎小鼠的药理作用及蛋白质组学效应的实验研究

目的　实验观察山茱萸提取物 (FCE)对慢性肾炎小鼠的治疗作用。方法　用右旋糖酐 (Dextran)作为抗原 ,诱发小鼠IgA肾炎 ,再用FCE进行实验性治疗 ,观察IgA肾炎小鼠尿蛋白含量、血清尿素氮 (BUN)和肌酐 (SCr)的含量 ,肾脏做病理组织检查 ,并使用双向电泳 (2 DE)技术提取分离小鼠肾脏总蛋白 ,分析、比较双向电泳图谱 ,寻找正常组与模型组、治疗组小鼠肾脏蛋白质组学差异。结果　小鼠注射抗原后 ,出现蛋白尿 ,血清BUN、SCr出现异常升高 ,用FCE治疗 ,显著降低了尿蛋白含量 (P <0 0 1)。FCE给药按 10g·kg-1和 5g·kg-1剂量能明显降低IgA肾炎小鼠血液中BUN和SCr含量 (P <0 0 1) ,并有一定利尿作用。病理组织检查发现FCE对Dextran造成的肾小球病变有保护作用 ,免疫荧光显微镜下可以观察到在IgA肾炎小鼠肾脏系膜区有IgA沉积。 2 DE获得较好的小鼠肾脏双向电泳图谱 ,并观察到治疗组与对照组有差异蛋白 ,如蛋白点 2 10 5 (Mr=2 8 0 32 ,pI=5 710 1)、2 10 8(Mr=2 8 0 15 7,pI =5 7383)在正常组弱表达 ,而在模型组高表达 ,给予FCE治疗后 ,相对含量下调 ,趋于正常 ;蛋白质点32 19(Mr=2 3 2ku ,pI=6 2 ) ,32 1(Mr=2 5 3ku ,pI=6 1)仅出现于正常组而未在模型组出现。结论　FCE具有改善肾功能、缓解肾脏损伤的     

原发性免疫球蛋白A肾病患者临床、病理研究

目的 探讨原发性免疫球蛋白A肾病[Primary Immunoglobin A nephropthy,IgAN(IgA肾病)]牛津分类四项病理指标与临床预后的关系.方法:回顾性分析157例IgA肾病患者的临床、病理资料.根据IgA肾病牛津分类建议的四个病理指标(肾小球系膜细胞增生、肾小球内皮细胞增生、节段性硬化或粘连、...     

ONO-AE-248诱发中性粒细胞非凋亡性程序化死亡的蛋白质组学分析方法的建立

目的:建立双向电泳技术和质谱技术在探索ONO-AE-248诱发的中性粒细胞(neutrophil,PMN)非凋亡性程序化死亡的分子机理研究中的应用,为深入认识“非凋亡性程序化死亡“这种新的死亡方式提供新的有效的研究途径.方法:将采用梯度离心法分离的,人正常PMN,分为正常对照组和实验组,实验组用ONO-AE-248刺激12小时建立非凋亡性坏死模型,提取对照组和实验组全蛋白,双向电泳分离蛋白质,采用PDQuest 2-DE软件分析找出两组间差异表达的蛋白质斑点,用基质辅助激光解吸电离飞行时间串联质谱(MALDI-TOF-MS)进行鉴定.结果:双向电泳图谱显示:ONO-AE-248诱导的人PMN非凋亡性程序化细胞死亡与PMN的自发性凋亡的蛋白质组存在差异显著,质谱鉴定初步得到其中12种差异表达的蛋白质.结论:双向电泳和质谱鉴定技术可以有效的分离和分析ONO-AE-248诱发的PMN非凋亡性坏死的蛋白质组,为进一步探索“非凋亡性程序化死亡“方式提供了新的方法和途径,为丰富细胞死亡方式带来了新的前景.     

Origin of urinary fibrin/fibrinogen degradation products in glomerulonephritis.

To elucidate the origin of the fibrin/fibrinogen degradation products (F.D.P.) occurring in the urine in glomerulonephritis 28 patients with glomerulonephritis were examined for renal fibrinolytic activity, F.D.P. in urine and serum, and blood fibrinolytic activators and blood fibrinolytic activators and inhibitors. Unlike the glomerful of healthy kidneys, which were fibrinolyticly inactive, those of kidneys with glomerulonephritis constantly showed fibrinolytic activity. The presence or absence of fibrin in the glomeruli was almost always accompanied by, respectively, the presence or absence of urinary F.D.P., which suggested a renal origin of urinary F.D.P. in glomerulonephritis. The low fibrinolytic activity of the blood and the absence of F.D.P. in the serum of these patients make it unlikely that the urinary F.D.P. in glomerulonephritis result from glomerular filtration.     

Renal toxicity after chronic inhalation exposure of rats to trichloroethylene

Male Long-Evans rats were exposed to 0 (controls) or 500 ppm trichloroethylene (TRI) for 6 months, 6 h daily, and 5 days a week. The TRI metabolites trichloroethanol (TCE) in blood and trichloroacetic acid (TCA) in urine were measured. Specific parameters related to the renal damage were determined in urine [biomarker for glomerular damage: high molecular weight proteins (HMW), albumin (ALB); for proximal tubular damage: N-acetyl-beta-D-glucosaminidase (NAG), low-molecular-weight-proteins (LMW)]. Significantly increased concentrations of NAG and LMW in urine of exposed rats were detected. No DNA-strand breaks in kidney cells could be detected using the comet assay, and histological examinations were performed. Histological alterations were observed in glomeruli and tubuli of exposed rats. The release of biomarkers for nephrotoxicity suggested alterations preferably in the proximal tubules of the exposed rats.     

Serum beta2-microglobulin in the assessment of renal function.

This study was undertaken to compare the serum beta2-microglobulin (beta2-m) as a test of renal function with the plasm creatinine and the glomerular filtration rate as estimated by the 24 hour endogenous creatinine clearance and the single injection 51Cr EDTA clearance method. Of the 33 patients with a variety of renal diseases and the four healthy volunteers studied, an excellent correlation was found between the serum beta2-m concentration (measured by radioimmunoassay) and the plasma creatinine, the creatinine clearance and the 51Cr EDTA clearance. When a more simple and less expensive method becomes available for the measurement of serum beta2-m it could prove a useful test of renal function. The assay of beta2-m in the urine could prove valuable for assessing whether proteinuria is glomerular or tubular in origin.     

Renal digoxin clearance: Dependence on plasma digoxin and diuresis

Summary The renal handling of digoxin in animals involves glomerular filtration, tubular secretion and tubular reabsorption, while only glomerular filtration and tubular secretion have been described in humans. The influence of plasma digoxin and urine flow on the renal handling of digoxin was investigated in 6 healthy volunteers. Non-glomerular renal excretion of digoxin (tubular secretion minus tubular reabsorption) was inversely correlated with plasma digoxin concentration and directly with urine flow. Hence, the present study demonstrated the occurrence of tubular reabsorption in addition to glomerular filtration and tubular secretion of digoxin. The results suggest that renal clearance of digoxin should be increased by increased urine flow, which might be of importance during digoxin toxicity.     

Proteomic Analysis of Colonic Mucosal Tissue from Tuberculous and Ulcerative Colitis Patients

Changes in the expression profiles of specific proteins leads to serious human diseases, including colitis. The proteomic changes related to colitis and the differential expression between tuberculous (TC) and ulcerative colitis (UC) in colon tissue from colitis patients has not been defined. We therefore performed a proteomic analysis of human TC and UC mucosal tissue. Total protein was obtained from the colon mucosal tissue of normal, TC, and UC patients, and resolved by 2-dimensional electrophoresis (2-DE). The results were analyzed with PDQuest using silver staining. We used matrix-assisted laser desorption ionization time-of-flight/time-of-flight spectrometry (MALDI TOF/TOF) to identify proteins differentially expressed in TC and UC. Of the over 1,000 proteins isolated, three in TC tissue and two in UC tissue displayed altered expression when compared to normal tissue. Moreover, two proteins were differentially expressed in a comparative analysis between TC and UC. These were identified as mutant β-actin, α-enolase and Charcot-Leyden crystal protein. In particular, the expression of α-enolase was significantly greater in TC compared with normal tissue, but decreased in comparison to UC, implying that α-enolase may represent a biomarker for differential diagnosis of TC and UC. This study therefore provides a valuable resource for the molecular and diagnostic analysis of human colitis.     

男女新生儿脐带血血清蛋白质组的比较

为发现不同性别新生儿之间蛋白质表达谱的差异,利用胰蛋白酶对上述血清蛋白分别进行酶解,并通过反相高效液相色谱法(RP-HPLC)对肽段混合物进行了分离,进一步通过电喷射离子化串联质谱法(ESI-MS/MS)鉴定了分离的肽段。本研究中共鉴定出837种非冗余蛋白质,其中不同类型血清中共有的蛋白质有282种,另有239种和213种分别属于男性新生儿和女性新生儿特有蛋白质。对其中53种差异蛋白质进行了分析,初步探讨了它们在胎儿生长发育以及性别分化过程中的作用。     

Acute and subacute nephrotoxicity of 2-bromophenol in rats

The present report aims at providing broader information on the acute nephrotoxicity of 2-bromophenol (2-BP) (a bromobenzene (BB) metabolite), due to its action on the kidneys, after repeated administration. Investigations were performed on female rats. Following a single dose, the most pronounced changes involved: concentrations and rates of excretion of proteins in urine, the number of epithelial cells excreted in urine, creatinine and urea clearance and reduced glutathione in renal tissue. Immediate effects could be ascribed to both renal tubules and glomeruli, mirrored in the level of urinary proteins and intensified excretion of renal epithelial cells. Less pronounced changes of the indicator values were noted under repeated dosing of 2-BP. The results obtained in a single exposure study confirm earlier reports on the mild nephrotoxicity of 2-BP following exposure to high doses. However, the transition from single to repeated exposure does not result in enhanced nephrotoxicity.