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。     

尿Ⅳ型胶原浓度与IgA肾病肾组织损害的关系

目的:观察IgA肾病(IgAN)患者尿Ⅳ型胶原(Col-Ⅳ)浓度及其与肾组织Col-Ⅳ表达和肾脏病理之间的关系,以期寻找一种能反映IgAN肾损害的非创伤性临床检测指标.方法:ELISA法测定IgAN患者血、尿Col-Ⅳ浓度;免疫组化法检测肾组织Col-Ⅳ表达;应用计算机病理图像分析系统对肾小球基质基底膜面密度、小球细胞个数及肾小管间质病变程度进行半定量分析,观察尿Col-Ⅳ浓度与IgAN患者肾组织Col-Ⅳ表达及肾病理损伤的关系.结果:IgAN患者尿Col-Ⅳ浓度、肾组织Col-Ⅳ表达较健康人明显增高,尤其是增生硬化和间质纤维化显著者更为明显;尿Col-Ⅳ浓度与肾组织Col-Ⅳ表达、肾小球基质基底膜面密度、小管间质损害显著正相关,且在轻度细胞外基质积聚和小管间质损害时,尿Col-Ⅳ浓度即增高;与小球内细胞个数呈负相关,而与肾小球系膜区、毛细血管襻区免疫球蛋白IgA、IgG、IgM和C3沉积的量和沉积范围无关,与血Col-Ⅳ水平无显著相关.结论:IgAN患者尿Col-Ⅳ浓度明显增高,尿Col-Ⅳ水平可作为监测IgAN患者早期肾硬化的一项指标.     

白细胞介素17在IgA肾病肾小管间质的表达及意义

目的观察白细胞介素17（IL-17）在IgA肾病患者肾组织的表达,探讨IL-17在IgA肾病中的临床及病理意义。方法选择51例IgA肾病患者肾组织活体检查采用Katafuchi半定量积分标准和免疫组织化学技术进行病理学积分和检测IL-17的表达,同时检测51例IgA肾病患者中30例外周血单个核细胞（PBMCs）中IL-17、转化生长因子β1（TGF-β1）、IL-6、IL-23、IL-βmRNA的表达。结果正常肾组织中未见IL-17表达。51例IgA肾病患者中,21例肾小管上皮细胞胞质IL-17表达阳性,2例肾间质淋巴细胞IL-17表达阳性,总阳性率为45．10％;并且随着肾小管间质病变加重,IL-17的表达逐步增多。IgA肾病患者PBMCs中IL-17、IL-6、IL-23、IL-1βmRNA的表达较正常人显著升高（P〈0．01）,TGF-β1mRNA的表达与正常人比较无统计学差异（P〉0．05）,IL-17mRNA与TGF-β1、IL-β1mRNA的表达呈正相关（r=0．67、0．71,P〈0．05）。IgA肾病患者肾小管间质IL-17的表达水平与尿位相畸形红细胞计数、肾小管间质病理积分呈正相关（r=0．60、0．67,P〈0．05）,IL-17mRNA的表达与血肌酐呈正相关（r=0．66,P〈0．05）。结论IgA肾病患者肾小管间质及外周血中IL-17表达增高,并与临床及病理预后指标密切相关,提示IL-17可能是参与IgA肾病发病及进展的因素之一。     

活化白细胞黏附分子在IgA肾病中的表达及意义

目的:探讨活化白细胞黏附分子(CD166)在IgA肾病中的表达及与IgA肾病临床病理特征的关系.方法:用免疫组化二步法检测CD166蛋白在79例IgA肾病患者肾组织的表达.结果:CD166在IgA肾病患者肾小管、间质普遍表达,按Lee分级的级别增加逐渐增高.与单纯性血尿患者(A组)相比,混合蛋白尿、血尿患者(B组)肾小管间质CD166表达明显增高.肾小管间质CD166表达与患者病理分级、小管间质病变、单个核细胞浸润程度及血清C反应蛋白(CRP)水平正相关.结论:CD166在IgA肾病中表达异常,在IgA肾病小管、间质病变及疾病进展中起一定作用.     

转化生长因子β1和结缔组织生长因子在狼疮肾炎肾小管间质中的表达

目的 研究狼疮性肾炎肾小管间质转化生长因子β1（TGF－β1）和结缔组织生长因子（CTGF）的表达特点及其意义。方法 应用免疫组织化学方法比较原发性肾小球肾炎微小病变型（MCD）和狼疮肾炎（LN）肾小管间质中TGF－β1和CTGF的表达及分布以及浸润细胞，α－平滑肌肌动蛋白（SMA）的表达特点。结果 TGF－β1及CTGF可不同程度地表达于LN的肾小球和肾小管间质，且较MCD的表达普遍增高，肾间质TGF－β1表达量与CTGF的表达量之间的相关系数r=0.5316,P=0.023。LN患者肾间质内CD3阳性细胞，CD68阳性细胞，PCNA阳性细胞以及α－SMA的表达明显高于MCD患者。α－SMA的表达程度与肾小管间质纤维化程度存在正相关关系，r=0.436,P=0.032。结论 LN肾间质内TGF－β1和CTGF表达增高，可能与LN肾间质病变及炎性细胞的浸润有关。     

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。     

CD34在IgA肾病肾小管间质病变中的表达及意义

目的:研究白细胞分化抗原CD34在IgA肾病肾小管间质病变中的表达变化,探讨其在IgA肾病进展过程中的作用.方法:应用免疫组织化学双标记技术检测不同小管间质病变程度的IgA肾病患者肾组织内CD34和α-平滑肌激动蛋白(α-SMA)的表达.肾小管间质病变分级采用Katafuchi积分.详细收集每例患者肾活检时的24 h尿蛋白定量(TUPr)及内生肌酐清除率(Ccr),并与免疫病理结果进行相关分析.结果:无小管间质病变组未见CD34的表达,α-SMA仅表达于间质的动脉壁;轻度小管间质病变组CD34和α-SMA表达散在,主要见于小管间质受损部位;中、重度病变组较轻度病变组CD34和α-SMA的表达显著增加(P＜0.05);小管间质内CD34的表达与α-SMA的表达及TUPr、Ccr具有显著相关性(P＜0.05).结论:随小管间质病变进展而出现的CD34表达增多可能在IgA肾病中发挥重要作用.     

正常人与原发性肾小球肾炎及狼疮性肾炎患者血尿IL-18水平比较

目的:探讨IL-18在原发性肾小球肾炎(PGN)及狼疮性肾炎(LN)发生、发展中的作用,以及寻找有助于两类疾病的鉴别诊断和对肾组织炎症活动程度进行评估的指标。方法:应用酶联免疫吸附检测(enzyme-linkedimmunosorbent assay,ELISA)法测定16例正常人、21例原发性肾小球肾炎(PGN)患者和18例LN患者血浆和尿液IL-18水平的变化。结果:LN患者血浆及尿液IL-18水平显著高于正常对照组(P均<0.001)和PGN组(P均<0.05),而且WHOⅣ型LN患者血浆及尿IL-18水平均明显高于非Ⅳ型LN患者(P<0.05);PGN患者尿液IL-18水平也高于正常人(P<0.05),但血浆IL-18水平与正常人比较无统计学差异(P>0.05);LN患者血浆IL-18水平与SLEDAI呈正相关(P<0.01),而尿IL-18水平与狼疮性肾炎RHSAI是密切正相关(P<0.001),但尿IL-18水平与血浆IL-18水平相关性没有统计学意义(P>0.05)。结论:IL-18参与LN的全身免疫病理过程,但可能仅参与PGN肾组织局部炎症过程;血浆IL-18检测可能有助于区分LN和PGN,尿IL-18的检测可望作为一项估计LN肾组织病变活动程度的有用指标。     

Wnt/β-catenin在IgA肾病中的表达及意义

目的:探讨Wnt/β-catenin及其下游靶基因在IgA肾病中的表达,分析其与患者的临床病理参数的相关性。方法:根据肾小管间质病变程度分组,采用RT-PCR、Western blot和En Vision-免疫组化技术检测各组IgA肾病患者以及正常对照组患者肾活检组织中Wnt/β-catenin及其下游靶基因snail、基质金属蛋白酶-7(MMP-7)、α-平滑肌动蛋白(α-SMA)的表达,各组之间进行比较。结果:IgA肾病患者肾活检组织中β-catenin、snail、MMP-7、α-SMA mRNA和蛋白表达水平较正常对照组升高(P0.05)。β-catenin、snail在伴有轻度肾小管间质病变的IgA肾病患者肾活检组织中表达水平高于无肾小管间质病变的IgA肾病患者(P0.05),其后随着肾小管间质病变程度的加重逐渐下降。MMP-7、α-SMA在IgA肾病患者肾活检组织中的表达水平与肾小管间质病变的严重程度相一致。结论:Wnt/β-catenin及其下游靶基因在IgA肾病中表达异常,可能参与IgA肾病的发生发展。     

非胰岛素依赖性糖尿病肾病患者肾小管间质病变的观察

观察了２６例非胰岛素依赖性糖尿病（ＮＩＤＤＭ）肾病患者肾小管间质病变的特点，包括肾小管功能的改变，肾组织形态学改变及部分患者的尿表皮生长因子（ＥＧＦ），结果发现，１．ＮＩＤＤＭ肾病小管间质功能受损较为弥漫，既有近端肾小管细胞损伤，也有远端小管的损伤，在形态这上表现为肾小管萎缩，基底膜增厚，间质扩张，弥漫炎细胞浸润及不同程度的纤维，２．在肾小球滤过功能明显受损之前，小管间质病变即已出现，并随着肾功能     

Increased excretions of beta2-microglobulin, IL-6, and IL-8 and decreased excretion of Tamm-Horsfall glycoprotein in urine of patients with active lupus nephritis

Tubulointerstitial nephritis is a less frequently recognized but important complication of systemic lupus erythematosus. We have investigated the cytokine beta2-microglobulin (beta2M) and Tamm-Horsfall glycoprotein (THG) excretions in the urine of systemic lupus erythematosus patients to identify indices for evaluation of tubulointerstitial inflammation in lupus nephritis (LN). Daily urine was collected from 15 patients with active LN, from 12 patients with inactive LN, and from 17 normal subjects. The amounts of soluble interleukin (IL) 2 receptor, IL-6, IL-8, beta2M, and THG in urine were measured. Beta2M and THG were regarded as indicators of proximal and distal renal tubule function, respectively. The urinary excretions of IL-6 and IL-8 were significantly higher in patients with active LN than in those with inactive LN and in normal individuals. The excretion of soluble IL-2 receptor in all three groups of subjects was not significantly different. On the other hand, the excretion of beta2M in patients with LN was significantly higher than that in normal individuals. The excretion of beta2M in patients with active or inactive LN was not significantly different. The THG excretion was lower in patients with active LN and tubulointerstitial inflammation as compared with patients with inactive LN or normal individuals. Six patients underwent pulse cyclophosphamide therapy during the course of experiments. Five of them showed a decrease in IL-8 and IL-6 excretions in urine after the treatment. The excretions of beta2M and THG in urine, in addition to IL-6 and IL-8, can reflect the renal inflammatory activity in patients with lupus tubulointerstitial nephritis as well as in those having lupus glomerulonephritis.     

68例狼疮肾炎的肾小管间质病变探讨

目的探讨肾小管间质病变（ＴＩＬ）在狼疮肾炎（ＬＮ）的意义。方法分析了６８例ＬＮ资料，并对其中４２例作了较长期的随访。结果ＬＮ的ＴＩＬ发生率为７５％，ＴＩＬ明显者，肾小管功能显著下降，血肌酐、尿蛋白水平明显增加（Ｐ＜００１），肾小管间质与肾小球急、慢性病理改变之间均显著相关（Ｐ＜００１），同时，存在明显ＴＩＬ者，其生存率及肾存活率有下降趋势。结论ＴＩＬ与肾小球病变呈平行的正相关关系，且对ＬＮ预后有一定的影响。     

Gene expression of TWEAK/Fn14 and IP-10/CXCR3 in glomerulus and tubulointerstitium of patients with lupus nephritis

Aim: The role of the tumour necrosis factor‐like weak inducer of apoptosis (TWEAK)/Fn14 and interferon‐inducible protein (IP‐10)/CXCR3 axis in the pathogenesis of lupus nephritis were studied. Methods: The mRNA expression of TWEAK, Fn14, IP‐10 and CXCR3 were quantified in the glomerulus and tubulointerstitium of 42 patients with lupus nephritis (LN group) and 10 healthy controls. Results: As compared to controls, LN patients had higher glomerular expression of TWEAK and Fn14, but glomerular CXCR3 expression was lower in the LN group. Similarly, the LN group had higher tubulointerstitial expression of TWEAK and Fn14, but lower tubulointerstitial expression of CXCR3, than controls. Glomerular TWEAK expression of class V nephritis was significantly higher than class IV nephritis. Glomerular expression of CXCR3 significantly correlated with proteinuria (r = ?0.532; P = 0.019), whereas tubulointerstitial CXCR3 significantly correlated with serum creatinine (r = ?0.447; P = 0.029). Conclusion: In patients with lupus nephritis, there is an increase in intra‐renal expression of TWEAK and Fn14, and a decrease in CXCR3 expression. Intra‐renal expression of CXCR3 correlates with proteinuria and renal function. Our findings suggest that the TWEAK/Fn14 and IP‐10/CXCR3 axis may contribute to the pathogenesis of lupus nephritis.     

Effect of human anti-DNA antibodies on proximal renal tubular epithelial cell cytokine expression: implications on tubulointerstitial inflammation in lupus nephritis

This study aimed to investigate the effects of human anti-DNA antibodies (Ab) from patients with lupus on renal proximal tubular epithelial cells (PTEC), focusing on alterations in cell morphology and proinflammatory cytokine synthesis. Immunohistochemistry showed increased tubulointerstitial IL-6 expression and IgG deposition in renal biopsies from patients with diffuse proliferative lupus nephritis, not observed in controls or membranous lupus nephritis, which correlated with the severity of inflammatory cell infiltration. Sera from patients with lupus nephritis contained IgG that bound to cultured PTEC. Such binding increased with disease activity and correlated with the level of anti-DNA Ab. Incubation of PTEC with anti-DNA Ab that were isolated during active (active Ab) or inactive (inactive Ab) disease induced IL-6 synthesis, both apically and from the basolateral aspect. This was accompanied by altered cell morphology, increased cell proliferation (P < 0.05), and lactate dehydrogenase release (P < 0.05). The binding of inactive Ab and active Ab to PTEC resulted in differential and sequential upregulation of TNF-alpha, IL-1beta, and IL-6 secretion (P < 0.05). Early induction of TNF-alpha was observed with active Ab; the two then acted synergistically to induce IL-6 secretion. Exposure of PTEC to inactive Ab was associated with modest induction of TNF-alpha, which was not involved in downstream induction of other proinflammatory peptides. These data suggest distinct immunopathogenetic mechanisms during disease flare or remission. Conditioned media from human mesangial cells acted synergistically with anti-DNA Ab to induce cytokine secretion in PTEC. Results from these studies underscore the pivotal role of PTEC in the pathogenesis of tubulointerstitial inflammation and fibrosis in lupus nephritis.     

Glomerular and tubulointerstitial miR-638, miR-198 and miR-146a expression in lupus nephritis

Aim: MicroRNAs (miRNAs) play important roles in the pathogenesis of autoimmune diseases. We studied the intra‐renal expression of miRNA targets that were reported to be differentially expressed in peripheral blood or urine between lupus nephritis (LN) patients and normal controls. Methods: We quantified the expression of in glomerulus and tubulointerstitium of miR‐146a, miR‐155, miR‐198 miR‐638 and miR‐663 in 42 patients with LN and 10 healthy controls. Results: As compared with controls, LN patients had lower glomerular expression of miR‐638 (P < 0.001) but higher tubulointerstitial expression of this target (P = 0.001). Both glomerular and tubulointerstitial expression of miR‐198 were higher in LN patients than controls (P < 0.001). For miR‐146a, LN patients only had higher expression in glomerulus (P = 0.005) but not in tubulointerstitium. Tubulointerstitial miR‐638 expression was significantly correlated with proteinuria (r = 0.404; P = 0.022) and disease activity score (r = 0.454; P = 0.008), while glomerular miR‐146a expressions were correlated with estimated GFR (r = 0.453; P = 0.028) and histological activity index (r = 0.494; P = 0.027). Conclusion: We found that intra‐renal expression of miR‐638, miR‐198 and miR‐146a are differentially expressed between LN patients and normal controls. Furthermore, the degree of change in glomerular miR‐146a and tubulointerstitial miR‐638 expression correlated with clinical disease severity. The results suggested that these miRNA targets may play a role in the pathogenesis of lupus nephritis.     

狼疮肾炎患者血清和尿白细胞介素-18水平检测及其临床意义

目的：检测狼疮肾炎(LN)患外周血清和尿液白细胞介素—18(IL—18)水平并探讨其临尿意义。方法：血清和尿液IL-18含量采用酶联免疫吸附方法(ELISA)。结果：LN组血清IL—18水平显高于正常对照组(P＜0、01)，活动期LN血清IL—18水平显高于缓解期患(P＜0．01)3LN组尿IL—18水平显高于正常对照组(P＜0．05)，活动期LN患尿IL—18水平显高于缓解期患(P＜0．05)。LN患血清IL—18水平与SLEDAI、抗dsDNA抗体成正相关关系，与补体C3呈负相关关系，而与血白蛋白、血肌酐无相关关系；活动期LN患尿IL—18水平与狼疮肾组织活动性指数(AI)、24h尿蛋白排泄量均呈正相关关系。结论：LN血清和尿IL—18水平显增高，IL—18可能在LN的病理生理过程中起重要作用。LN血清和尿IL—18水平均与狼疮病情活动密切相关，可作为判断狼疮疾病活动性的候选参考指标。     

活动性狼疮肾炎患者外周血单个核细胞钙调神经磷酸酶活性的检测及白细胞介素-4表达变化

目的：检测活动性狼疮肾炎（LN）患者外周血单个核细胞（PBMC）白细胞介素-4（IL-4）表达及其与钙调神经磷酸酶（Calcineurin，CaN）活性的关系。方法：体外培养活动性LN患者PBMC，应用发色底物法检测胞浆CaN活性，逆转录PCR检测IL-4mRNA表达。结果：（1）在单纯培养情况下，正常对照组和LN组PBMC均出现一定量CaN活化，活动性LN组显著高于正常对照组[（46．08±5．58）mmol／mg．pr vs（8．81±3．61）mmol／mg．pr，P〈0．01]；在PMA＋Ionomyci刺激下，各组CaN活性均升高，活动性LN组CaN活性明显高于正常对照组[（69．34±12．59）mmol／mg．pr vs（37．12±11．57）mmol／mg．pr，P〈0．01]；（2）LN患者PBMC在单纯培养和PMA＋Ionomycin刺激时IL-4蛋白和mRNA表达均显著高于相应的对照组（P〈0．05）；（3）在单纯培养和PMA＋Ionomycin刺激时，FK506对LNPBMC表达IL-4蛋白和mRNA均有显著抑制作用（P〈0．01）。结论：LN患者PBMC存在CaN过度活化；LN患者PBMC高效表达IL-4与其CaN过度活化密切相关，通过阻断CaN活性可调控IL-4表达。     

淋巴细胞功能相关抗原1共刺激作用在活动性狼疮肾炎中的作用

目的探讨淋巴细胞功能相关抗原(LFA)1及其信号分子在狼疮肾炎发病机制中的作用。方法 利用免疫沉淀、免疫印迹和RTllPCR技术检测了LFA-1共刺激对19例活动性狼疮肾炎(LN)患者外周血单个核细胞(PBMC)P13-K磷酸化产物、IL-10 mRNA表达的影响。以14例健康献血员作为对照。结果 在CD3 mAb30 ng/ml存在的情况下,LFA-1mAb共刺激明显提高了活动性LN患者PBMC P13-K磷酸化产物表达(P<0.01),伴有IL-10 mRNA表达和蛋白含量增加(P<0.01),抗lFA-1抗体(5μmol/L)加入后明显抑制了P13-K磷酸化产物表达(P<0.01)。在CD3 mAb和LFA-1mAb协同刺激的情况下,P13-K特异抑制剂Wortamannin的加入明显抑制了LN患者PBMCIL-10 mRNA水平和蛋白含量(P<0.01)。结论 LFA-1作为共刺激分子过度活化P13-K可能介导了活动性狼疮肾炎IL-L 10的高效表达。