IgA肾病肾组织α—平滑肌肌动蛋白的表达

ＩｇＡ肾病肾组织α－平滑肌肌动蛋白的表达陈振斌梁平郑智勇余英豪庄永泽谢福安孙慧敏陈莲云肾小球系膜区损伤是许多肾小球疾病包括ＩｇＡ肾病（ＩｇＡＮ）的中心环节，其中系膜细胞具有不同的表型及许多功能，我们对此进行探讨，并观察肾间质α－平滑肌肌动蛋白（ＳＭＡ...     

低亲和力钠依赖二羧基转运蛋白随增龄表达变化的研究

目的 研究大鼠及人类肾脏低亲和力钠依赖(Na~+/)二羧基转运蛋白(NaDC1)随增龄的表达变化规律并探讨其在肾脏衰老变化中的意义。方法 采用Northern杂交、Western印迹及免疫组化等方法对大鼠出生后1d、7d、1个月、3个月、12个月、24个月及少年、中青年、老年正常人肾脏 NaDC1的表达变化。结果 大鼠NaDC1 mRNA表达呈现随肾组织发育成熟表达逐渐增强,1个月达高峰,后随增龄表达下降的趋势(P<0.05)。Western印迹显示NaDC1蛋白随鼠龄增高表达逐渐增强,3个月达高峰,后表达渐降的趋势。免疫组化结果显示大鼠及人类肾组织NaDC1分别表达于近端小管刷状缘,大鼠生后1d表达最弱,后表达渐强,到3个月达高峰(5.30±1.52比1.40±0.43,P<0.01),后呈现渐降的趋势(P<0.05)。人类肾组织NaDC1表达呈现随增龄渐降的趋势。结论 大鼠及人肾组织进入衰老时,NaDC1 mRNA及蛋白表达均呈现下降的趋势,可作为对肾脏衰老观察的指标之一。     

人钠/二羧酸协同转运蛋白3基因调控肾小管上皮细胞线粒体膜电位的研究

目的 观察人钠／二羧酸协同转运蛋白3(hNaDC3，)对人肾脏近曲小管上皮细胞(HKC)线粒体膜电位的变化及其对细胞能量代谢的影响。方法 应用亚克隆技术构建正义pcDNA3-hNaDC3和反义pcDNA3-AhNaDC3两个真核表达载体，通过脂质体LipofectAMINE将pcDNA3-hNaDC3及pcDNA3-AhNaDC3转染至HKC细胞。克隆筛选后，用RT—PCR、Northern印迹及Western印迹鉴定外源基因的整合和表达。荧光探针JC-1观察各细胞系线粒体膜电位的变化。结果 外源hNaDC3基因稳定整合到HKC细胞基因组中，并获得高、低表达。转染正义hNaDC3cDNA的HKC细胞线粒体膜电位降低，JC-1在线粒体内形成单体，发出绿色荧光；而转染反义hNaDC3cDNA的HKC细胞线粒体膜电位略微升高，JC-1形成聚合体，发出红色荧光。结论 hNaDC3过表达引起线粒体膜电位降低，反义hNaDC3则使线粒体膜电位略微升高。提示NaDC3可能通过使线粒体膜电位下降，参与了细胞能量代谢。     

载脂蛋白H在肾病综合征患儿肾组织表达的研究

目的研究载脂蛋白H(ApoH)在原发性肾病综合征(PNS)患儿肾组织中的分布及表达变化,探讨其在PNS进展过程中的作用。方法利用荧光定量RT-PCR和免疫组化检测78例不同病理类型的PNS患儿肾组织ApoH的mRNA及蛋白表达,并对肾组织中的肾小管损伤、间质炎症细胞浸润进行评分。肾活检前检测血白蛋白(Alb)、血脂、Scr、尿视黄醇结合蛋白(RBP)、尿蛋白定量。14例正常肾组织为对照。结果(1)正常和PNS患儿肾组织中均有ApoH蛋白表达,主要分布于皮质近肾小球的近端肾小管上皮细胞;正常及PNS患儿肾组织均有ApoHmRNA表达,且肾组织ApoH蛋白与mRNA表达呈显著正相关,r=0.264,P<0.05,提示肾脏本身能够合成ApoH.(2)不同病理类型PNS患儿肾组织ApoHmRNA及蛋白表达存在差异,微小病变NS(MCNS)、膜性肾病(MN)组ApoHmRNA及蛋白表达水平分别为4.95±0.40、4.73±0.60和12.06±2.04、12.35±0.61,与对照组(5.44±1.56和12.69±1.89)相比虽有减少,但差异无统计学意义,P>0.05;系膜增生性肾小球肾炎(MsPGN)、局灶性节段性肾小球坏死(FSGS)组ApoHmRNA及蛋白表达水平分别为3.30±0.28、2.82±0.36和10.13±3.09、10.12±1.02,明显低于MCNS、MN及对照组,差异均有统计学意义,P<0.05;(3)激素耐药组ApoHmRNA表达低于激素敏感组,分别为4.27±0.30、     

IgA肾病患者肾组织中增殖细胞核抗原表达

用免疫组化方法研究了２０例原发性ＩｇＡ肾病患者肾组织中增殖细胞核抗原（ＰＣＮＡ）表达。结果显示ＩｇＡ肾病患者肾组织中ＰＣＮＡ表达增加，肾小球内ＰＣＮＡ阳性细胞数、肾小管和间质中ＰＣＮＡ阳性细胞百分数均与肾组织学损害程度呈正相关；临床－病理研究显示肾小管中ＰＣＮＡ阳性细胞百分数分别与２４小时尿蛋白量、血清肌酐浓度（Ｓｃｒ）呈正相关；肾间质中ＰＣＮＡ阳性细胞百分数亦与２４小时尿蛋白量呈正相关。肾脏细胞增殖程度作为一项判断肾小球肾炎组织学损害程度和预后的指标值得进一步研究。     

大鼠低亲和力钠依赖二羧酸转运蛋白对枸橼酸及草酸转运电生理特性的研究

目的 探讨大鼠低亲和力钠依赖二羧酸转运蛋白(SDCT1)对枸橼酸及草酸的转运特点。方法克隆出大鼠SDCT1全长cDNA基因。应用爪蟾卵母细胞异源性表达SDCT1，并使用双电极电压钳法记录通道电流。通过改变灌流液中枸橼酸、草酸的浓度，以及两种底物转运时钠离子的浓度及pH值，对其特性进行了研究。结果SDCT1对枸橼酸及草酸的转运均为底物浓度及钠离子依赖。pH值变化显著影响2者的转运。但低pH对草酸的转运影响要远大于枸橼酸。结论在临床实践中，虽然升高尿液pH值可以抑制枸橼酸的重吸收，发挥其抑制钙盐沉积的功能。但过度地碱化尿液也会使草酸的重吸收受到抑制，使草酸大量存留在尿中而导致结行。保持适当的尿酸碱度是必要的。     

原发性IgA肾病肾组织及血中转化生长因子-β1表达的相关性

目的：探讨转化生长因子-β1（TGF-β1）在肾组织的表达与原发性IgA肾病（IgAN）各病理类型关系及其与血中的TGF-β1之间的关系。方法：分析了2003年1月～2008年6月的67例原发性IgAN患者肾组织中TGF-β1的表达与其各病理类型关系,及其与血中TGF-β1之间的关系。结果：IgA肾病患者各病理分级肾组织中TGF-β1的表达量差异有统计学意义（P〈0.01）,随病理分级增高,TGF-β1表达量越多。IgA肾病患者肾组织中TGF-β1的表达量与血中TGF-β1显著相关（r=0.918,P〈0.01）。结论：原发性IgA肾病患者可通过检测血中的TGF-β1来反映肾组织中TGF-β1水平,从而预测其肾脏病理等级,有助于预测其预后,避免反复肾活检。     

原发性肾病综合征患者IL-18的血浆水平及其在肾组织的表达

目的：初步探讨白细胞介素18(IL—18)在原发性肾病综合征(PNS)发生发展中的作用。方法：采用酶联免疫吸附(ELISA)法测定11例正常人及24例PNS患者血浆IL—18水平，同时用免疫组织化学方法检测6例正常肾组织和上述24例PNS患者肾组织IL—18的表达量。结果：PNS患者血浆IL—18水平与正常对照组IL较无统计学意义；而且各种病理类型间的差异也没有统计学意义(P均＞0．05)；而肾小球及肾小管—间质IL—18表达量却均显著高于正常对照组(P均＜0．01)。不同病理类型PNS肾小球区IL—18表达量存在差异，以膜增生性肾小球肾炎(MPGN)表达量为最高，其次为系膜增生性肾小球肾炎(MesPGN)，而膜性肾病(MD)、局灶节段性肾小球硬化(FSGS)和轻微病变(MCD)的表达量则相对较低，并且肾小球区IL—18表达量与24h尿蛋白排泄量(24h　UPQ)至正相关，与血浆白蛋白浓度(Alb)至负相关(r分别为0．669和－0．727，P均＜0．01)；肾小管—间质区IL—18表达量与小管—间质损害程度至正相关(r＝0．484，P＜0．05)。结论：肾组织IL—18高表达可能参与PNS的发病过程，而又可能以自分泌或/和旁分泌方式起作用。     

CTGF在IgA肾病肾组织中的表达与临床病理关系

20世纪60年代,免疫荧光技术的发展让我们认识了IgA肾病,40年后的今天,长期的临床观察、大量的基础研究,让我们更加重视这一疾病。目前,人们正致力于寻找有效生物标志物并进行整合以从发病机制的不同环节预测IgA肾病的进展和预后,结缔组织生长因子（CTGF）已成为这一研究领域中新热点。     

肾结石大鼠钠/二羧基转运蛋白1的表达及枸橼酸钾防治机制的研究

目的 研究肾组织钠／二羧基转运蛋白1（SDCT1）与低枸橼酸尿的关系以及枸橼酸钾的干预作用，探讨肾结石发病的分子机制和防治措施。方法 雄性Wistar大鼠分为对照组、肾结石组及枸橼酸钾干预组。血、尿枸橼酸和草酸采用酶法测定，Northern blot检测大鼠肾组织SDCT1mRNA水平的改变，免疫组织化学观察SDCT1在肾组织的分布及表达变化。结果 与对照组比较，肾结石组第3天尿草酸水平显著升高，枸橼酸水平显著降低，同时肾组织SDCT1mRNA及其蛋白水平上调。第7天SDCT1mRNA及其表达产物增加更为显著，同时尿枸橼酸水平进一步降低，尿钙排泄显著增加，87.5%大鼠有中－大量的草酸钙结石形成。第14天上述改变更为明显，结石形成率达100％。枸橼酸钾干预组各时间点尿草酸水平与肾结石组差异无显著性意义，但尿枸橼酸水平显著高于肾结石组及对照组，肾组织SDCT1mRNA及蛋白表达显著低于肾结石组，与对照组差异无显著性意义；结石形成率显著低于肾结石组；肾小管扩张、炎细胞浸润等病变也明显减轻。结论 肾组织SDCT1表达上调可能是低枸橼酸尿的重要原因，与肾结石的形成有密切关系。枸橼酸钾可下调肾结石大鼠肾组织SDCT1的表达，对肾结石的形成具有明显的干预作用。     

IgA serology in recurrent and non-recurrent IgA nephropathy after renal transplantation

BACKGROUND: This study investigated whether abnormal circulation of macromolecularIgA and IgA with altered glycosylation or electrical chargeplays a role in the recurrence of IgA nephropathy (IgAN) aftertransplantation. STUDY DESIGN: A total of 92 renal transplant patients were enrolled; 52 IgANpatients and 40 with other non-IgAN. The IgAN group included10 patients showing IgA mesangial deposits in the grafted kidneys(recurrent group) and 10 who did not (immunohistochemicallyproven non-recurrent group). In addition another 22 IgAN transplantpatients were clinically free of recurrent disease. METHODS: The analyses included macromolecular IgA (IgAIC) detected bythe conglutinin assay (K), heavy IgA precipitated in 2.5% polyethyleneglycol (PEG), IgA-fibronectin aggregates (IgA/F Aggr), mixedIgA/IgGIC, IgA binding to mesangial matrix components (fibronectin,laminin, type IV collagen) or polycations (poly-L-lysine) andIgA with altered glycosylation (Jacalin-binding assay). RESULTS: After transplantation, IgAN patients displayed significantlyhigher mean levels for each variable measured than non-IgAN(ANOVA, P <0.05). By stepwise regression analysis, the bindingof IgA to fibronectin had the highest coefficient. By comparingdata in recurrent and clinically non-recurrent IgAN, we observedthat two groups could be distinguished by the results of thetwo assays for macromolecular IgA (conglutinin IgAIC and IgA-fibronectinaggregates) and IgA with increased affinity for type IV collagen(P <0.05). When the selected group of immunohistochemicallyproven non-recurrent IgAN was compared to the recurrent one,a statistically significant difference was found only for thebinding of IgA to type IV collagen (P<0.05). Data from thistest were significantly related with proteinuria (P<0.05)and microscopic haematuria (P <0.04). CONCLUSION: Even though the IgA serology of renal transplant IgAN patientsshows peculiar features and recurrent and non-recurrent IgANdiffer in many aspects, the prevalence of positive data in thetwo groups had no predictive value. This suggests that the recurrenceof IgAN is modulated by factors affecting the interaction betweencirculating abnormal IgA and mesangial cells and/or matrix.     

转铁蛋白受体在IgA肾病肾组织中的表达

目的阐明转铁蛋白受体（cD71、TfR）在IgA肾病（IgAN）肾组织中表达的分布特点及其与IgA表达的关系,探讨其在IgAN免疫发病机制中的作用。方法120例肾活检患者根据临床表现和肾活检病理诊断分为原发性IgAN组（原发组）44例、非IgAN性IgA沉积组（继发组）38例、无IgA沉积肾病组（肾病组）38例。用免疫荧光双套色法标记的抗体,在激光共聚焦荧光显微镜下观察四甲基罗丹明（TRITC）标记的CD71和异硫氰荧光素（FITC）标记的IgA在肾组织上的表达。结果CD71的表达强度与IgA的沉积程度相一致。CD71与IgA在原发组肾小球上高表达;在继发组低表达;在对照组微弱表达或不表达。激光共聚焦荧光显微镜下观察到CD71表达与IgA表达呈现共位现象。结论CD71在IgAN患者肾小球呈现高表达,其表达与IgA分子呈现共位状态,提示CD71可能参与了IgAN免疫发病过程。     

Recurrence of IgA nephropathy in renal transplants

SUMMARY: The present article provides an update from the literature on the clinical significance of recurrent IgA disease in renal transplant patients, and it identifies possible reasons for the differences between the data that have been published. the clinical features of recurrent IgA nephropathy are described here, along with suggestions for the management of these patients.     

肾组织中乙型肝炎病毒抗原阳性和阴性IgA肾病的临床与病理对照

目的比较ＩｇＡ肾病（ＩｇＡＮ）肾组织中乙型肝炎病毒抗原（ＨＢＡｇ）阳性和阴性患者的临床与病理特点。方法收集经免疫组化检测证实的肾组织ＨＢＶ抗原阳性组２６例和阴性组５９例ＩｇＡＮ患者的临床和病理及随访资料。结果与阴性组相比，阳性组肾病综合征者多见，血红蛋白、血ＩｇＡ水平和肌酐清除率显著下降，免疫荧光以ＩｇＡ＋ＩｇＧ＋ＩｇＭ型者多见，肾组织病理损害以Ⅳ级为多，肾小球硬化、肾小管病变、间质炎症和纤维化程度均显著严重。两组在临床表现、实验室检查、病理改变及预后方面均有显著性差异，阳性组临床病理改变更严重、预后更差。结论两组是性质相差较大的病变，对阳性组进行抗病毒治疗尤为重要     

Pathogenesis of idiopathic IgA nephropathy

Despite a prodigious amount of work on the physiology of IgA production in man, and many studies on the immunopathology of IgA nephropathy, ranging from the immunogenetics to the immune response to chemical characteristics of the IgA, we are hardly any nearer to defining the pathogenesis of this disease. One of the main changes in our understanding has been to recognise that the bone marrow, now known to produce normally one-third of the body's IgA, overproduces this immunoglobulin in IgA nephropathy. This alters the previous notion that IgA nephropathy was due simply to IgA production in the mucosa, although a mucosal component is not excluded. Certain characteristics of the IgA in the diseased kidney and the circulation have been defined: it is of subclass IgA1 and has a higher proportion of light chains and negative charge than in normal subjects. The specificities of the IgA, either in the kidney or in complexes, have not helped to clarify the pathogenesis. They have been found for a wide range of endogenous and exogenous antigens, suggesting that the antibody activity represents polyclonal B cell activation. These findings have not helped to confirm the prevailing theory that IgA nephropathy is an immune complex disease. Other theories put forward are that IgA nephropathy is an autoimmune disease, glomerular components or IgA itself being among the candidate antigens, or that there is primary dysregulation of the IgA immune system. At this stage of development in our understanding of this common nephropathy, it is important to guard against the assumption that idiopathic IgA nephropathy is one disease and is the result of a single pathogenetic mechanism.     

Recurrent IgA nephropathy after renal transplantation despite immunosuppressive regimens with mycophenolate mofetil.

BACKGROUND: Transplantation offers an excellent option for patients with immunoglobulin-A nephropathy (IgAN) with severe renal dysfunction. However, IgAN frequently recurs in allografts treated with azathioprine. We examined the impact of mycophenolate mofetil immunosuppression on recurrence of IgAN. METHODS: We reviewed the charts of patients transplanted for IgAN at our institution in the cyclosporin era. Patients were excluded from further analysis if follow-up was <12 months or if immunosuppression at engraftment did not include azathioprine or mycophenolate mofetil. Laboratory data, medications and allograft biopsy findings were compiled. RESULTS: 152 kidney transplantations met the study criteria. At engraftment, 61 allografts were treated with azathioprine and 91 with mycophenolate mofetil. By 3 years post-transplant, IgAN developed in six of 60 (10.0%) azathioprine-treated allografts and five of 62 (8.1%) mycophenolate mofetil-treated allografts (P = 0.76). Overall, 13 azathioprine-treated and seven mycophenolate mofetil-treated allografts showed recurrence. As expected in this retrospective study, the duration of observation was longer in the azathioprine group. The interval between engraftment and diagnosis of recurrent disease was also longer. Survival of allografts with recurrent IgAN was similar in the two groups. Survival of allografts with recurrent IgAN was worse than for allografts without recurrence or allografts transplanted into patients with non-IgAN renal failure. Neither switching azathioprine to mycophenolate mofetil nor using an angiotensin-converting enzyme inhibitor or angiotensin-II type 1 receptor blocker ameliorated the clinical course after a biopsy documented recurrent IgAN. CONCLUSIONS: Mycophenolate mofetil, compared with azathioprine, did not lessen the recurrence of IgAN or its clinical impact.