尿巨噬细胞移动抑制因子浓度与狼疮肾炎肾组织损害的关系

目的观察狼疮肾炎(LN)患者尿巨噬细胞移动抑制因子(MIF)浓度是否升高及其与LN肾组织MIF表达、肾脏病理及功能损害的关系,试图寻找一种反映狼疮肾损害的非创伤性指标。方法用ELISA方法测定LN患者血、尿MIF浓度。用免疫组织化学双染技术观察肾组织MIF表达及巨噬细胞浸润情况。了解尿MIF水平与狼疮肾组织MIF表达、巨噬细胞浸润、狼疮肾组织活动指数、肾脏功能及组织学损害的关系。结果LN患者肾组织MIF表达及尿MIF浓度较正常人明显增高,尤以增生及炎症明显的Ⅲ、Ⅳ型LN为明显,尿MIF浓度与肾组织MIF表达、巨噬细胞浸润、狼疮肾组织活动指数、肾小管损害显著相关,但与血MIF水平、蛋白尿程度及肾功能损害无显著相关。结论LN患者尿MIF浓度明显增高,并与肾组织MIF表达及狼疮肾组织活动情况显著相关,可作为一种监测狼疮肾活动及肾损害的非创伤性指标。     

核因子-κB在狼疮肾炎肾组织中的表达及其意义

目的了解狼疮肾炎(LN)肾组织中核因子(NP)-κB的表达并探讨其与LN肾脏病理改变和肾功能损害的关系.方法以NF-κB亚基P65单抗为抗体,采用微波免疫组织化学染色(APAAP法)检测LN肾组织NF-κB表达,并进一步分析其与肾小球内C-myc蛋白表达、LN活动指数、肾脏病理和功能损害的关系.结果狼疮肾炎肾组织中NF-κB表达较正常肾组织显著增高,以WHOⅣ型为最显著.NF-κB在肾小球和肾小管均有表达,但小管表达更显著.LN肾组织NF-κB阳性细胞数与肾小球C-myc蛋白表达量、肾组织活动指数、肾脏病理改变和肾功能损害显著相关.结论NF-κB可能参与了LN的发病机制,肾组织中NF-κB的表达可作为反映狼疮肾组织活动病变和进行性肾损害的参考指标.     

骨调素对巨噬细胞在新月体肾炎肾脏组织局部浸润的影响

目的观察骨调素（ＯＰＮ）在大鼠实验性新月体肾炎肾脏组织中的表达及功能性抑制ＯＰＮ的活性对巨噬细胞在肾脏局部浸润的影响。方法ＯＰＮ在肾脏组织蛋白和基因水平的表达分别应用免疫组化双标记及原位杂交技术。结果在实验性新月体肾炎肾脏组织内ＯＰＮ的表达明显升高，并与肾脏局部巨噬细胞的浸润、肾脏病理损害程度及肾功能的下降明显相关。应用抗ＯＰＮ抗体治疗以后可显著抑制ＯＰＮ在肾脏组织的表达，同时巨噬细胞在肾脏组织局部的浸润也明显减少，肾脏病理损害减轻、肾功能明显改善。结论ＯＰＮ的高表达是实验性新月体肾炎发病过程中巨噬细胞在肾脏组织局部浸润的重要介导因子，抑制ＯＰＮ的表达和功能可显著抑制肾脏局部巨噬细胞的聚集及改善肾功能。     

核因子—kB在狼疮肾炎肾组织中的表达及其意义

目的：了解狼疮疮肾炎（ＬＮ）肾组织中核因子（ＮＦ）－ｋＢ的表达并探讨其与ＬＮ肾脏病理改变和肾功能损害的关系。方法以ＮＦ－ｋＢ亚基Ｐ６５单抗为抗体,采用微波免疫组织化学染色（ＡＰＡＡＰ法）检测ＬＮ肾组织ＮＫ－ｋＢ表达,并进一步分析其与肾小球内Ｃ－ｍｙｃ蛋白表达、ＬＮ活动指数、肾脏病理和功能损害的关系。结果狼疮肾炎肾组织中ＮＦ－ｋＢ表达较正常肾组织显著增高,以ＷＨＯⅣ型为最显著。ＮＦ－ｋＢ在肾小球和肾     

狼疮肾炎患者肾组织CD134(OX40)表达的免疫组织化学分析

目的：了解狼疮肾炎(LN)肾组织中CD134(OX40)的表达并探讨其与LN肾脏病理改变和肾功能损害的关系。方法：用免疫组织化学方法对40例LN患肾组织CD134的表达进行检测，并对其与LN的肾脏病理改变和肾功能损害的相关性进行分析。结果：LN患肾组织中，CD134表达显上调，尤其以WHO Ⅳ型LN更为明显。除系膜细胞、内皮细胞和远曲小管CD134表达明显增强外，肾间质毛细血管和大血管内皮细胞亦有CD134表达阳性的浸润细胞。LN肾组织加以表达与LN肾脏病理活动指数和肾功能损害显相关。结论：LN患确实存在CD134共刺激分子的异常表达，这在LN的发生和发展中可能起着重要的作用。     

细胞周期抑制蛋白p21 cipl在狼疮肾炎肾组织中的表达及其意义

目的 了解狼疮肾炎（LN）中细胞周期抑制蛋白p21^cip1的表达并探讨其与肾脏病病理改变，细胞增生及LN肾组织活动性的关系。方法 采用微波免疫组织化学染色法检测LN肾组织p21^cipl表达，并进一步分析其与肾小球增殖细胞核抗原（PCNA）阳性细胞数、LN活指数、细胞增生等肾脏病理改变的相关关系。结果 正常肾小球细胞无或仅有p21^cipl表达，LN肾小球细胞p21^cipl的表达显著上调。尤以非Ⅳ型为显著，LN肾组织p21^cipl阳性细胞数，PCNA阳性细胞数及LN肾组织活动指数显著负相关，结论 p21^cipl在LN肾小球细胞中呈高表达，且与肾小球细胞增生程度密切相关，p21^cipl可能参与LN发病的过程。     

白介素-1受体桔抗剂对实验性新月体肾炎骨调素表达的调节作用

目的探讨白介素-1受体拮抗剂（IL-lra）对实验性新月体肾炎肾脏组织骨调素（OPN）表达的调节作用，进一步阐明IL-1在实验性新月体肾炎发病机制中的作用。方法加速型实验性新月体肾炎模型应用兔抗鼠肾小球基底膜肾毒血清制备。第1组为模型组：注射肾毒血清后第0～7天，不加任何干扰治疗；另外2组为治疗组及治疗对照组：从第7～14天，分别给予IL-lra和生理盐水持续静脉点滴。结果模型组OPN的表达显著增高。生理盐水治疗组，OPN在肾小球和小管-间质细胞的表达更高，并和巨噬细胞的局部浸润及肾功能密切相关。而IL-lra治疗组，OPN的表达显著下调，巨噬细胞浸润明显减少，部分逆转肾功能。结论应用IL-lra抑制IL-1的活性可显著下调骨调素的表达和保护肾功能，提示IL-lra的治疗作用可能是通过抑制骨调素的表达，减少巨噬细胞在肾脏局部浸润的机制。     

CD44及其配体骨调素在大鼠实验性新月体肾炎中的表达及作用

目的探讨ＣＤ４４及其配体骨调素（ＯＰＮ）在实验性新月体肾炎发展过程中的表达和作用。方法应用兔抗鼠肾毒性血清制作新月体肾炎模型，免疫组化双标记及原位杂交技术检测ＣＤ４４、ＯＰＮ及其它指标。结果ＣＤ４４和ＯＰＮ在新月体肾炎的发展过程中表达明显升高，并和巨噬细胞在肾脏组织的局部浸润、新月体的形成及肾功能的损害密切相关。结论在实验性新月体肾炎模型，ＣＤ４４和ＯＰＮ在肾小球实质细胞和小管间质细胞显著表达，并可能在新月体肾炎的发病过程中发挥重要的作用。     

颗粒膜蛋白的表达与狼疮肾炎的关系研究

目的 了解血浆黏附分子颗粒膜蛋白（ＧＭＰ－１４０）水平及其在肾组织的表达与狼疮肾炎（ＬＮ）活动、肾脏病理及痛功能损害的关系。方法 用酶联免疫吸附法（ＥＬＩＳＡ）测定血浆ＧＭＰ－１４０水平,并用免疫组织化学染色法,观察正常肾和狼疮组织ＧＭＰ－１４０表达。结果 ＬＮ患者较正常人以及活动ＬＮ较非活动ＬＮ,血ＧＭＰ－１４０水平显著升高;ＬＮ肾组织ＧＭＰ－１４０表达较正常肾组织明显增强,且狼疮肾组织ＧＭＰ－     

乙型肝炎病毒相关性肾炎患者肾组织Notch1受体表达与临床病理的相关性

目的 观察Notch1受体在乙型肝炎病毒相关性肾炎（HBV-GN）患者肾组织中的表达分布特点,探讨其与肾组织病理及临床表现的相关性。 方法 以2008年至2010年经肾活检确诊的HBV-GN患者48例为研究对象。用免疫组化方法观察HBV-GN肾组织中Notch1受体的分布特征;免疫荧光双标观察Notch1受体与HBsAg的分布关系;分析其与病理类型、肾小球病变、肾小管病变及临床指标的关系。 结果 Notch1受体主要分布在肾小管上皮细胞及间质区域,呈棕红色颗粒状,肾小球内也有少量表达。HBV-GN组患者肾组织Notch1受体阳性积分显著高于血清HBsAg阳性或阴性的原发性肾小球肾炎患者及正常肾组织对照。系膜增生性肾小球肾炎（MsPGN）组和膜增生性肾小球肾炎（MPGN）组Notch1受体阳性积分较高,但各病理类型组间Notch1积分差异无统计学意义。Notch1受体分布与HBsAg分布一致,其强度与肾间质纤维化（r = 0.473,P = 0.001）、小管萎缩（r = 0.690,P = 0.000）、炎细胞浸润（r = 0.616,P = 0.000）等肾小管间质病变呈正相关;患者肾功能与Notch1受体阳性积分呈负相关（r = -0.393,P = 0.006)。 结论 HBV-GN患者肾组织Notch1受体的表达增强,主要分布在肾小管上皮细胞及间质,与HBsAg分布一致;其表达水平与肾间质病变和肾功能变化相关。Notch1受体表达异常可能参与了HBV-GN的肾脏病理的进展。     

Renal expression of matrix metalloproteinases in human ANCA-associated glomerulonephritis.

BACKGROUND: Expression of matrix metalloproteinases (MMPs) by infiltrating and intrinsic renal cells is increased in inflammatory conditions, and may correlate with disease activity of glomerulonephritis. We analysed renal expression of MMPs, tissue inhibitor of metalloproteinase-1 (TIMP-1) and markers of neutrophil and monocyte infiltration in renal biopsies of patients with active anti-neutrophil cytoplasmic antibody (ANCA)-associated glomerulonephritis. METHODS: Immunohistochemical expression of MMP-2, -3, -9, TIMP-1, the neutrophil- and monocyte-derived MMP activators cathepsin G, neutrophil elastase and myeloperoxidase (MPO), and the monocyte marker CD14 was determined in renal biopsies of active proteinase 3 (PR3)-ANCA (n = 7) and MPO-ANCA (n = 6) associated glomerulonephritis, and in normal renal tissue (n = 4). Double labelling experiments of MMPs and TIMP-1 were performed with MPO and CD68, labelling neutrophils and macrophages. RESULTS: MMP-2-, MMP-3-, MMP-9- and TIMP-1-positive cells were detected in ANCA-associated glomerulonephritis in glomeruli with active inflammation (cellular crescents or fibrinoid necrosis), only occasionally in normal appearing glomeruli, and not in sclerotic glomeruli and positive cells were found in the tubulo-interstitium. MMPs and TIMP-1 were expressed predominantly by MPO-and CD68-positive cells. In normal renal tissue, no expression was detected, with the exception of weak mesangial staining for MMP-2. In ANCA-associated glomerulonephritis, glomerular MMP-2, -9 and TIMP-1 correlated with glomerular cathepsin G expression, while the number of MMP-9-expressing cells per glomerulus correlated with the percentage of crescentic glomeruli. Tubulo-interstitial expression of MMPs correlated with all markers of neutrophil and monocyte infiltration, and interstitial MMP-9 and TIMP-1 expression correlated with renal function at the time of renal biopsy. CONCLUSIONS: Expression of glomerular and interstitial MMP-2, -3, -9 and TIMP-1 is increased in active ANCA-associated glomerulonephritis and correlates with inflammatory activity.     

Local macrophage proliferation correlates with increased renal M-CSF expression in human glomerulonephritis.

BACKGROUND: Macrophage accumulation is a prominent feature in many forms of glomerulonephritis. Local proliferation of macrophages within the kidney has been described in human and experimental glomerulonephritis and may have an important role in augmenting the inflammatory response. The current study examined the relationship between local macrophage proliferation and renal expression of macrophage colony-stimulating factor (M-CSF). METHODS: A total of 118 renal biopsies of patients with a wide range of glomerulonephridities were examined for M-CSF protein and macrophage proliferation (KP1+PCNA+cells) by single and double immunohistochemistry staining, respectively. RESULTS: Biopsies of thin membrane disease (TMD) with histologically normal kidney showed M-CSF protein expression by 33% of cortical tubules, while glomerular M-CSF expression was limited to resident macrophages and some podocytes. Glomerular M-CSF expression increased significantly in proliferative forms of glomerulonephritis, with M-CSF staining of infiltrating macrophages, podocytes and some mesangial cells. Segmental areas of strong M-CSF expression, particularly in crescents, co-localized with KP1+PCNA+ proliferating macrophages. There was also an increase in tubular M-CSF expression in most types of glomerulonephritis. Tubular M-CSF staining was strongest in areas of tubular damage and co-localized with KP1+ macrophages, including KP1+PCNA+ proliferating macrophages. Many interstitial macrophages and alpha-smooth muscle actin-positive myofibroblasts showed strong M-CSF staining. Statistical analysis showed a highly significant correlation between M-CSF expression and local macrophage proliferation in both the glomerulus and tubulointerstitium. Glomerular and tubular M-CSF expression gave a significant correlation with renal dysfunction. CONCLUSIONS: Glomerular and tubulointerstitial M-CSF expression is up-regulated in human glomerulonephritis, being most prominent in proliferative forms of disease. This correlated with local macrophage proliferation, suggesting that increased renal M-CSF production plays an important role in regulating local macrophage proliferation in human glomerulonephritis.     

白细胞介素13基因在人肾组织的表达及其意义

目的 探讨白细胞介素13（IL-13）基因在原发性非IgA系膜增生性肾炎（MsPGN）、原发性局灶节段性肾小球硬化症（FSGS）和狼疮肾炎（LN）患者肾组织的表达及意义。方法 采用原位杂交技术（ISH）检测34例上述3种肾脏病患者及4例对照组肾组织中IL-3 mRNA表达量的变化,分析IL-3基因表达量与肾功能变化的关系。结果 IL-13 mRNA在FSGS、MsPGN和LN患者肾组织内表达均增高,而对照者肾组织内无明显IL-3 mRNA表达。LN患者肾小管间质区IL-3 mRNA表达水平与狼疮活动相关,MsPGN及PSGS患者肾小管间质区IL-13 mRNA表达量与肾功能变化及24h尿蛋白量相关。结论 IL-13可能直接参与人类肾小球疾病的免疫反应过程。     

Anti-macrophage migration inhibitory factor reduces transforming growth factor-beta 1 expression in experimental IgA nephropathy.

BACKGROUND: In human glomerulonephritis, including immunoglobulin-A nephropathy (IgAN), glomerular expression of macrophage migration inhibitory factor (MIF) is found to correlate with progressive renal injury. We have shown previously that polymeric IgA is capable of inducing MIF production in cultured human mesangial cells, suggesting a role in inducing inflammatory injury in IgAN. Herein, we examined whether IgA deposition and the subsequent renal injury can be ameliorated with anti-MIF treatment in an experimental murine model of IgAN. METHODS: Glomerular IgA deposition was induced in 4-week-old BALB/c mice by intravenous injection of immune complexes consisting of dinitrophenyl-conjugated bovine serum albumin (DNP-BSA) and IgA MOPC-315 myeloma anti-DNP antibodies. To determine the therapeutic effect of anti-MIF, mice were given anti-MIF (5 mg/kg) or isotypic control antibody intravenously 2 h before the immune complexes administration. The mice were sacrificed 48 h after injection of DNP-IgA. Proteinuria and haematuria were determined and the kidneys were removed for histopathology, immunostaining and immunoblotting. The effect of exogenous MIF on production of TGF-beta 1 by cultured mesangial cells was also examined. RESULTS: IgA deposits were detected in glomeruli of all mice receiving the immune complexes while no glomerular deposit was detected in the control mice. Microscopic haematuria and mesangial hypercellularity were present in mice of the three experimental groups and were absent in the control group. Proteinuria was absent in all groups. Anti-MIF treatment also resulted in decreased renal expression of TGF-beta 1. Moreover, the reduction in TGF-beta 1 expression was confined mainly to glomerular mesangium. An in vitro culture experiment demonstrated that MIF increased TGF-beta 1 production in a time- and dose-dependent fashion. MIF-induced TGF-beta 1 synthesis was abolished by incubating cells with neutralizing antibody against MIF. CONCLUSIONS: Our finding shows that anti-MIF treatment can ameliorate kidney injury and reduce glomerular TGF-beta 1 expression in an experimental model of IgAN.     

Clinical significance of costimulatory molecules CD80/CD86 expression in IgA nephropathy

BACKGROUND: IgA nephropathy (IgAN) is the most common form of human glomerulonephritis. Tubulointerstitial inflammation with infiltration of mononuclear cells plays an important role in the progression of IgAN. Activation of T cells requires costimulatory signals through binding of CD28 receptor with cognate ligands (CD80/CD86) located on antigen-presenting cells (APC). To assess the clinical significance of this regulatory pathway participation in the pathogenesis of IgAN, a comprehensive immunohistologic evaluation was conducted on renal tissue of IgAN in different phases of progressive injury. METHODS: Thirty-three cases of IgAN and ten cases of non-IgA mesangial proliferative glomerulonephritis (PGN) with minor tissue damage as controls were investigated. Monoclonal antibodies were used to assess the expression of CD80, CD86, CD68, CD14, CD45RO, human leukocyte antigen-DR (HLA-DR), and intercellular adhesion molecule-1 (ICAM-1) in renal tissues. Clinical and expression data were compared at the time of renal biopsy. RESULTS: CD80+ and CD86+ cells were observed more in IgAN patients with progressive renal injury than in mild cases and controls. CD80 was limited to tubular epithelial cells and was complemented by HLA-DR expression. CD86 was expressed in the glomerulus, periglomerular area, and peritubular interstitium. Activated T cells (CD45RO+), monocytes (CD14+), macrophages (CD68+), and CD86 showed similar distributions. Positive correlations were found between CD86+ cells and CD45RO, CD14, and CD68 positive cells and between CD80+ tubuli and peritubular interstitial CD45RO+ cells. The number of interstitial CD86 positive cells and the percentage of CD80+ tubuli were correlated with renal function. Most CD86+ cells were monocyte/macrophages. CONCLUSION: This study suggested that CD80 and CD86 activate T cells in IgAN, CD80/CD86 expressions correlated with renal function at the time of renal biopsy, and monocyte/macrophages and tubular epithelial cells act as APC.     

Role of stem cell factor and mast cells in the progression of chronic glomerulonephritides.

BACKGROUND: Mast cells (MCs) have been implicated in the pathogenesis of atherosclerosis and tissue fibrosis. However, the role of MC in the development of renal fibrosis has not been fully elucidated. Stem cell factor (SCF; the ligand for MC c-kit receptor) is thought to attract and activate MCs. METHODS: The intensity of MC infiltration and SCF expression in renal biopsies from 56 patients with different forms of primary and secondary glomerulonephritis and five controls were investigated by immunohistochemistry, using a monoclonal anti-human MC tryptase antibody and a polyclonal antihuman SCF antibody. RESULTS: A large number of MCs were detected in the renal interstitium of the diseased kidneys. Immunostainable SCF was detected in tubular as well as interstitial cells. MC infiltration was significantly higher in glomerulonephritis (16.9 +/- 10.2 cells/field) compared with controls (2.8 +/- 2.1 cells/field, P = 0.03). Similarly, immunostainable SCF was 0.6 +/- 0.3% for controls and 3.3 +/- 2.1% in the glomerulonephritis group (P = 0.02). MC infiltration was highly correlated with SCF expression in diseased kidneys (r = 0.93, P = 0.0001). Double immunostain showed them to colocalize in some interstitial cells. Analysis of MC proliferation [proliferating cell nuclear antigen (PCNA) positivity] and apoptosis (in situ end labeling of DNA) showed these cells to be terminally differentiated. Both MCs and SCF were correlated with interstitial fibrosis (R = 0.71 for MC and R = 0.62 for SCF, P = 0.0001) and interstitial alpha-smooth muscle actin (R = 0.69 for MC and R = 0.60 for SCF P = 0.0001). Using regression analysis, the number of MC infiltration was found to be a very powerful determinant of interstitial fibrosis in the glomerulonephritis group (R2 = 91.4%). CONCLUSION: MCs as an infiltrating hematopoietic cell and its growth factor (SCF) seem to be up-regulated in glomerulonephritis, and may play a role in the development of renal fibrosis.     

Abnormal regulation of insulin-like growth factor gene expression in peripheral blood mononuclear cells from patients with IgA nephropathy.

We investigated insulin-like growth factor (IGF)-I and -II mRNA expression in peripheral blood mononuclear cells (PBMC) and T cells obtained from 31 patients with IgA nephropathy (IgAN), 43 patients with other types of glomerulonephritis and 16 health age-matched controls. The majority of patients with IgAN showed elevated IGF-I and -II mRNA expression in PBMC, while no IGF-I and -II mRNA expression was detected in PBMC obtained from patients with other types of glomerulonephritis or normal controls. In T cells obtained from IgAN, other types of glomerulonephritis and normal controls, however, IGF-I and -II mRNA expression was not detected. A positive correlation was noted between IGF-I and -II mRNA levels and urinary protein excretion. IGF-I and -II mRNA expression also correlated with the histopathological findings in the renal tissue of patients with IgAN. Sixty-nine percent of patients with more than 1.0 g/day proteinuria showed strong [more than (++)] IGF-I and -II mRNA expression in their PBMC. Eighty-one and 76% of patients with grade III and IV histopathological findings, respectively, showed strong IGF-I and -II gene expression in their PBMC. We also studied the clinical course of 11 patients with IgAN during hospitalization. The IGF-I and -II mRNA levels in these patients decreased gradually, as did proteinuria, after treatment. These studies suggest that abnormal regulation of IGF-I and -II gene expression in PBMC may be associated with the progression of IgAN and may be useful as an indicator of disease activity.     

肾小球疾病中转化生长因子-β_1与肌成纤维细胞的表达及意义

目的：探讨肾小球疾病中转化生长因子-β1（TGF-β1）与肌成纤维细胞的α-平滑肌肌动蛋白（α-SMA）表达及其对肾硬化的影响作用。方法：将原发性肾小球肾炎患者42例根据不同病理类型分为4组,利用肾活检组织免疫组织化学染色观察TGF-β1、α-SMA、胶原Ⅲ（Col-Ⅲ）表达与患者血肌酐（Scr）和内生肌酐清除率（Ccr）之间的相关性,根据镜下免疫组织化学染色在肾小球、肾小管间质中的着色深浅程度及分布,用MetuMorph图像处理系统,测出灰度按半定量方法,进行统计学分析。结果：不同病理类型的肾小球肾炎有不同的表达,随硬化程度的加重,其表达显著增加。TGF-β1表达主要分布于肾小球的系膜基质区、新月体中和小管间质的肾小管上皮细胞胞浆、间质成纤维细胞胞浆中、间质基质区和炎性细胞大量浸润区。α-SMA的表达在MsPGN组最高,而FSGS,SGN肾小球上几乎无α-SMA的表达,在肾间质中α-SMA的表达,随病变程度的加重,其表达显著增加。肾硬化中TGF-β1表达量与患者Scr和Ccr之间呈正相关,α-SMA、Col-Ⅲ在肾小管间质的表达量与患者Scr和Ccr之间呈正相关。结论：在肾硬化形成过程中,TGF-β1是重要的促发因素,肾硬化病变的初期,炎症反应使巨噬细胞和成纤维细胞等活化,分泌TGF-β1增加,TGF-β1进一步刺激（肾小管上皮细胞）TEC和成纤维细胞等,使表型转化为Myo-FB,Myo-FB合成和释放TGF-β1,进入恶性循环,最终导致肾硬化形成。