Monocyte chemoattractant protein-1 and interleukin-8 levels in children with acute poststreptococcal glomerulonephritis

The infiltration of leukocytes into the glomeruli is a major factor in inflammatory glomerular damage in acute poststreptococcal glomerulonephritis (APSGN). Chemokines participate in leukocyte infiltration. The aim of the present study was to investigate the role of monocyte chemoattractant protein-1 (CCL2/MCP-1) and interleukin-8 (CXL8/IL-8) in APSGN with special emphasis on their role in the clinical course of renal disease. Twenty-one children with APSGN were studied. Serum and urinary CCL2/MCP-1 and CXL8/IL-8 levels were measured by ELISA. The relationships between urinary chemokines and the degree of proteinuria were investigated. Serum and urinary CCL2/MCP-1 levels were significantly higher in the acute phase than in the resolution phase and in controls ( P <0.05). Urinary CCL2/MCP-1 levels in the control group were significantly lower than in both the acute and resolution phases ( P =0.01 and P =0.001, respectively). In the acute phase, urinary CCL2/MCP-1 correlated with the extent of proteinuria ( r =0.58, P =0.006) but not with serum CCL2/MCP-1 levels ( r =0.21, P =0.36). Urinary and serum CXL8/IL-8 levels were significantly elevated in the acute phase compared with the resolution phase and controls ( P <0.05). A consistent increase in urinary CCL2/MCP-1 was found in the acute phase of patients with APSGN, and this correlates with the degree of proteinuria. Our results emphasize the important role of locally produced chemokines in immune-mediated glomerular injury.     

Urinary monocyte chemoattractant protein-1 (MCP-1) is a marker of active renal vasculitis.

BACKGROUND: Macrophage infiltration and cytokine production are important in the pathogenesis of crescentic glomerulonephritis in anti-neutrophil cytoplasm antibody (ANCA)-associated vasculitis. The aim of this study was to investigate whether urinary levels of chemokines, monocyte chemoattractant protein-1 (MCP-1) and fractalkine, were useful tools for non-invasive assessment of renal vasculitis. METHODS: In a prospective study, concentrations of chemokines were measured in urine and serum samples using specific enzyme-linked immunosorbent assays, and related to the patients' clinical status. Renal expression of MCP-1 was studied by immunohistochemical staining of renal biopsies. RESULTS: Urinary levels of MCP-1 were significantly higher in patients with active (P<0.01) or persistent (P<0.05) renal vasculitis, in comparison with healthy volunteers, control patients, patients with inactive vasculitis and patients with extra-renal disease only. There were no differences in serum concentrations of MCP-1 between these groups. Reduction in urinary MCP-1 levels following treatment preceded the improvement of renal function by a median of 2 weeks. In one patient, rising urinary levels of MCP-1, despite immunosuppressive therapy, was associated with progression to severe renal failure. There were no differences in urinary fractalkine levels between the different groups of patients and controls. Immunohistology of renal biopsies from patients with crescentic glomerulonephritis showed increased staining for MCP-1 in glomerular and interstitial cells. Urinary MCP-1 levels correlated with glomerular, but not tubulointerstitial, macrophage infiltration (P<0.05). CONCLUSIONS: This study shows that measurement of urinary MCP-1, but not fractalkine, is a useful non-invasive technique for the assessment of renal involvement and monitoring the response to therapy in ANCA-associated vasculitis.     

单核细胞趋化因子-1在间质性膀胱炎中表达的研究

目的 探讨单核细胞趋化因子-1(MCP-1)在间质性膀胱炎(IC)患者膀胱组织和尿液中的表达水平及其意义. 方法根据美国国立糖尿病、消化、肾病协会IC诊断标准确诊女性IC患者35例,感染性膀胱炎(UI)患者20例,肾囊肿患者25例作为正常对照组.IC患者平均年龄47(31～65)岁.主诉下腹酸胀/疼痛和夜尿次数多,伴有尿频尿急;已生育30例.均行24 h排尿卡记录、IC症状及问题评分表(O'Leary-Sant评分表)、钾离子敏感试验(PST)和麻醉状态下水扩张后膀胱镜检查.膀胱镜检查获取3组患者膀胱组织和尿液样本,免疫组化染色方法观察MCP-1在IC组织中的表达分布.RT-PCR技术测定3组膀胱组织中MCP-1 mRNA表达水平,酶联免疫吸附试验测定3组尿液标本中MCP-1水平. 结果 IC、UI、对照组尿液标本中MCP-1浓度分别为(74.1±36.9)、(280.6±68.9)、(10.8±6.9)pg/ml;膀胱组织中MCP-1相对定量值分别为76.2±24.0、99.5±30.1、36.1±14.1.lC组和UI组中组织/尿液中MCP-1表达水平均高于正常对照组,差异有统计学意义(P＜0.01).IC组临床症状评分为(14.9±1.8)分,与MCP-1升高水平呈正相关(r=0.686). 结论 IC患者膀胱组织和尿液中MCP-1表达升高,可能成为IC诊断的非特异性免疫指标之一.     

The role of monocyte chemoattractant protein-1 in biliary atresia

Background

The aim of this study was to explain the role of monocyte chemoattractant protein-1 (MCP-1) in biliary atresia (BA).

Methods

Concentrations of serum MCP-1 and collagen type IV were measured in 38 patients with BA by using commercially available kits. MCP-1 was also assessed in liver biopsy specimens by using immunohistochemistry. Subjects were classified into groups. Group 1 comprised BA patients with normal liver function (n = 13), group II comprised BA patients with moderate liver dysfunction (n = 18), group III comprised BA patients older than 20 years awaiting liver transplantation (n = 7), and the control group comprised age-matched patients without evidence of liver disease (n = 23).

Results

Serum MCP-1 levels were significantly increased in group II compared with group I (P < .0001) and the control group (P < .0001). Serum MCP-1 levels in group III were lower than in the control group (P < .0001). There was a significant linear correlation between serum MCP-1 levels and type IV collagen levels in group II. Group II subjects with portal hypertension (PH) had higher MCP-1 levels than those without PH (P = .0009). Biopsy specimens showed MCP-1 was expressed mainly on biliary epithelial cells, vascular endothelial cells, and hepatocytes in group II.

Conclusions

These findings suggest that MCP-1 probably plays a significant role in the development of progressive liver fibrosis in BA.     

Olmesartan inhibits the expression of monocyte chemoattractant protein-1 and tumor necrosis factor-α and improves vascular remodeling after vascular injury in mouse

Vscularinjuryinducesmigrationofsmoothmusclecells (SMCs)fromthemediatotheintimaandsubsequent proliferationofSMCswithintheintima .Thesevascularresponsesareproposedtobecriticalprocessesofneointimalformationandinducenarrowingofthevascularlumenandarteriosclerosis .1,3Manystudieshavedemonstratedthatangiotensinconvertingenzyme(ACE)inhibitors4 ,5inhibitneointimalformationintheinjuredmodelofsmallanimal,whereasotheragentsthathaveblood pressuredeclinedfailtopreventit.4 Ininjuredarteries,theexpression…     

Increased expression of monocyte chemoattractant protein-1 (MCP-1) by renal epithelial cells in culture on exposure to calcium oxalate, phosphate and uric acid crystals.

BACKGROUND: During the development of non-infectious kidney stones, crystals form and deposit in the kidneys and become surrounded by monocytes/macrophages (M/M). We have proposed that in response to crystal exposure renal epithelial cells produce chemokines, which attract the M/M to the sites of crystal deposition. We investigated the expression of monocyte chemoattractant protein-1 (MCP-1) mRNA and protein by NRK52E rat renal tubular epithelial cells exposed to calcium oxalate (CaOx), brushite (Br, a calcium phosphate) and uric acid (UA) crystals. METHODS: Confluent cultures of NRK52E cells were exposed to CaOx, Br or UA at a concentration of 250 micro g/ml (66.7 micro g/cm(2)). They were exposed for 1, 3, 6, 12, 24 and 48 h for isolation of mRNA and 24 h for ELISA to determine the secretion of protein into the culture medium. Since cells are known to produce free radicals on exposure to CaOx crystals we also investigated the effect of free radical scavenger catalase on the crystal induced expression of MCP-1 mRNA and protein. RESULTS: Exposure of NRK52E cells to the crystals resulted in increased expression of MCP-1 mRNA and production of the chemoattractant. CaOx crystals were most provocative while UA the least. Treatment with catalase had a negative effect on the increased expression of both MCP-1 mRNA and protein, which indicates the involvement of free radicals in up-regulation of MCP-1 production. CONCLUSION: Exposure to both CaOx and calcium phosphate crystals stimulates increased production of MCP-1. Free radicals appear to be involved in this up-regulation. Results indicate that MCP-1, which is often associated with localized inflammation, may be one of the chemokine mediators associated with the deposition of various urinary crystals in the kidneys during kidney stone formation. Because of the small number of experiments performed here, results must be confirmed by more extensive studies with larger sample size.     

MCP-1-2518位点基因多态性对脊柱结核易感性的影响

目的:分析趋化因子单核细胞趋化蛋白-1(MCP-1)启动子区2518基因型频率和等位基因多态性与中国湖南省汉族人群脊柱结核易感性的关系.方法:连续收录2004年12月～2012年8月我院收治的湖南汉族新发脊柱结核患者及健康志愿者,应用聚合酶链式反应(PCR)和DNA直接测序法检测两组患者MCP-1-2518位点的多态性,运用R×C卡方检验比较两组基因型分布频率的差异,计算各比值比(0R)及95％可信区间(CI).结果:共纳入符合条件的脊柱结核患者334例,健康志愿者336例.MCP-1-2518GG、AG、AA 三种基因型在病例组和对照组中的分布频率分别为37.1％、50.3％、12.6％和25.6％、54.2％、20.2％,两组组间比较有显著性差异(P＜0.05),其中MCP-1-2518GG基因型的分布频率在脊柱结核人群中显著增高.MCP-1基因-2518 G/A多态位点G、A等位基因频率在病例组为62.3％和37.7％,在对照组为52.7％和47.3％,两组等位基因的分布频率存在统计学差异(P＜0.05),-2518G等位基因与脊柱结核发病相关,其OR值为1.483,95％ CI为1.193～1.844.结论:在湖南汉族人群中,MCP-1 2518位点GG基因型和G等位基因均可能与脊柱结核的易感性相关.     

单核趋化蛋白-2中和抗体对骨癌痛大鼠脊髓背角磷酸化细胞外信号调节蛋白激酶表达的影响

目的 观察鞘内注射单核趋化蛋白-2(monocyte chemoattractant protein-2,CCL2)中和抗体后骨癌痛大鼠行为学的变化,并探讨其可能的镇痛机制. 方法 雄性SD大鼠32只,体重180～220 g,采用随机数字表法分为4组(每组8只):假手术+正常IgG组(I组),于大鼠左侧胫骨干骺端骨髓腔内注射10μl Hank's液;假手术+CCL2中和抗体组(Ⅱ组),于大鼠左侧胫骨干骺端骨髓腔内注射10μl Hank's液,在注射Hank's液后第7～9天鞘内注射CCL2中和抗体,每天1次,每次10μl,浓度为103 mg/L;骨癌痛+正常IgG组(Ⅲ组),于左侧胫骨干骺端骨髓腔内注射10 μl(1×107/ml)Walker 256肿瘤细胞;骨癌痛+CCL2中和抗体组(Ⅳ组),在注射肿瘤细胞后第7～9天鞘内注射CCL2中和抗体,每天1次,每次10μl,浓度为103 mg/L.观察造模前及术后1、3、6、7、8、9d大鼠机械缩足反射阈值(mechanical withdraw threshold,PMWT)和热缩足反射潜伏期(thermal withdraw latency,TWL)及脊髓背角磷酸化细胞外信号调节蛋白激酶1/2(phosphorylate extracellular signal-regulated kinases 1/2,p-ERK1/2)的表达. 结果 与Ⅰ组比较,Ⅲ组大鼠的PMWT和TWL在第6～9天明显下降,脊髓背角p-ERK1/2蛋白表达明显增加(P＜0.01);与Ⅲ组比较,Ⅳ组大鼠的PMWT和TWL在第6～9天明显上升(P＜0.01),脊髓背角p-ERK1/2蛋白表达明显下降(P＜0.01);Ⅰ组和Ⅱ组上述指标比较,差异无统计学意义(P＞0.05). 结论 鞘内注射CCL2中和抗体可以部分缓解骨癌痛大鼠的机械性和热痛觉超敏,这种效应可能与抑制p-ERK1/2的表达有关.     

中药黄芪对脊髓星形胶质细胞单核细胞趋化蛋白-1分泌的影响

目的 :研究黄芪对体外培养的脊髓星形胶质细胞趋化蛋白 1(MCP 1)分泌的影响。方法 :自Wistar大鼠脊髓组织分离、纯化星形胶质细胞 ,体外培养 ,分别予以TNF α及TNF α 黄芪处理 ,ELISA方法检测培养上清液中MCP 1的表达。结果 :TNF α可以显著刺激星形胶质细胞合成、释放MCP 1,而黄芪可下调其刺激作用。结论 :受炎症因子刺激活化的星形胶质细胞可能是损伤脊髓局部MCP 1的来源之一 ,黄芪可以减少脊髓损伤后内源性MCP 1的产生 ,从而缓解继发性脊髓损伤 ,发挥脊髓保护作用。     

Premature senescence and cellular phenotype transformation of mesangial cells induced by TGF-B1

Abstract

Background: Transforming growth factor-β1 (TGF-β1) is a polypeptide member of the transforming growth factor β superfamily of cytokines and performs many cellular functions. Its overexpression may lead to renal fibrosis. Aim: This study planed to investigate the effects of TGF-β1 on the cell cycle and phenotype of mesangial cells. Methods: Rat mesangial cells were cultured together with different concentrations (0, 1, 2, 5, and 10?ng/mL) of TGF-β1 for specified times from 0?min to 72?h. 0?ng/mL TGF-β1 and 0?min served as controls. Cell cycles were assessed by flow cytometry and α-smooth muscle actin expression (α-SMA) protein expression by western blot analysis. All data were presented as Mean?±?SD. Statistical analysis was performed by using one-way analysis of variance and correlation analysis. Results were considered significant at p?<?0.05. Results: After 15?min of co-culture with different concentrations of TGF-β1, the percentage of mesangial cells in G0/G1 phase was significantly elevated compared to the control (p?<?0.05). 12?h co-culture induced cell hyperplasia, 24?h co-culture obvious up-regulation of α-SMA (p?<?0.01) and one or two cells’ myofibroblast phenotype transition, and 36?h co-culture several cells’ phenotype transition. Correlation analysis prompted that the TGF-β1-induced premature aging was time-dependent (p?<?0.01). Conclusion: TGF-β1 may induce mesangial cells’ premature senescence and myofibroblast-like phenotype transformation time-dependently, which may contribute to the development of early stage of glomerulosclerosis.     

p38丝裂素活化蛋白激酶信号通路在单核细胞趋化蛋白1介导系膜细胞增殖及细胞外基质表达中的作用

目的探讨p38丝裂素活化蛋白激酶（MAPK）信号通路在单核细胞趋化蛋白1（MCP-1）介导大鼠系膜细胞（MCs）增殖及纤连蛋白（FN）和Ⅰ型胶原（ColⅠ）表达中的调控作用。方法用四甲基偶氮唑盐比色法（MTT法）评估不同浓度的MCP-1（12．5、25、50、100ng／ml）及p38MAPK阻断剂SB203580（1、5、10μmol／L）于不同时间对体外培养的大鼠MCs的增殖作用。采用逆转录多聚酶链反应（RT—PCR）检测细胞内FN、ColⅠmRNA的表达。用ELISA法检测上清液FN、ColⅠ蛋白含量。结果MCP-1能刺激MCs的增殖，呈剂量和时间依赖性（P〈0．05），而p38MAPK阻断剂明显抑制上述MCP-1的作用（P〈0.01）。MCP-1能使细胞FN、ColⅠ的表达上调，而p38MAPK阻断剂能抑制其上调表达的作用。结论p38MAPK信号转导通路在MCP-1介导大鼠MCs增殖及FN和ColⅠ表达中起调控作用。MCP-1在系膜增生性肾小球肾炎（MsPGN）的发病中起一定的作用。     

Synergistic action of inflammation and lipid dysmetabolism on kidney damage in rats

In kidney disease, inflammation and lipid dysmetabolism are often associated together, however, the effect and mechanism of inflammatory mediators and lipid dysmetabolism on kidney damage is still unclear. In this study, Wistar rats were randomized into four groups: normal diet?+?saline (Group N), high-fat diet (HF)+?saline (Group HF), normal diet?+?adriamycin (Group ADR), HF?+?adriamycin (Group ADR?+?HF). After 10?weeks of feeding, rats in each group were randomly sacrificed. We found that the protein content of urine in ADR and ADR?+?HF groups were significantly higher than that of group N and HF while the serum levels of total protein and albumin in the ADR and ADR?+?HF groups decreased correspondingly. The serum levels of triglyceride, total cholesterol and low-density lipoprotein in the HF, ADR and ADR?+?HF groups increased. In the treatment groups, mesangial proliferation, matrix accumulation, tubular vacuolization, inflammatory cell infiltration and fat deposition were detected. These pathological changes were the most serious in the ADR?+?HF group. The expression of tumor necrosis factor-α (TNF-α) and transforming growth factor-β1 (TGF-β1) were increased in each treatment group, especially in the ADR?+?HF group. Our results suggested that the inflammatory factors and abnormal lipid levels can activate the inflammatory response in kidney of the Wistar rats, and lead to a series of pathological changes in renal tissue, and inflammatory factors and lipid dysmetabolism can aggravate damage in the kidney.     

MCP-1在大鼠坐骨神经损伤中作用的实验研究

目的 研究单核细胞趋化蛋白-1(MCP-1)在大鼠坐骨神经损伤后在神经组织中的表达以及抑制其作用对损伤的坐骨神经华勒变性过程的影响.方法 96只雄性SD大鼠随机分为4组A、B、C、D(n=24),另取42只随机分为2组E、F组(n=21),于右大腿后部中段切断坐骨神经,A、B组用聚乙烯软管套接固定神经远端于充满MCP-1抗体的微渗透泵直到取材,近端返折固定于临近肌肉.C、D、E、F组神经切断后远端旷置,近端处理同A组.A、B组于0 h、24 h、3 d、7 d(n=6)分别作光镜和电镜观察远端神经轴突和髓鞘形态变化,C、D组分别为其对照.E、F组在损伤后0 h、6 h、12 h、24 h、3 d、7 d、14 d(n=3)取坐骨神经远端,E组实时荧光定量RT-PCR测定MCP-1 mRNA表达,F组Western Blot测定MCP-1蛋白含量.结果 神经损伤后,在24 h和14 d时E组MCP-1 mRNA和F组MCP-1蛋白水平达到高峰,于24 h时最高.光镜和电镜下各实验组髓鞘厚度均高于各自对照组.结论 MCP-1在大鼠坐骨神经切割伤后在时间上存在分泌高峰,在神经损伤后远端华勒变性过程中起促进作用.     

Urinary levels of epidermal growth factor, interleukin-6 and monocyte chemoattractant protein-1 may act as predictor markers of renal function outcome in immunoglobulin A nephropathy

Aim:   Urinary cytokine excretion may reflect histological changes in immunoglobulin A nephropathy (IgAN), and their measurement can give information about disease outcome.
Methods:   Thirty-three IgAN patients were prospectively followed for 5.6 ± 3.1 years. Urinary levels of monocyte chemoattractant protein-1 (MCP-1), interleukin (IL)-6 and epidermal growth factor (EGF) were measured at diagnosis and repeated 1 year later for IL-6 and EGF.
Results:   Urinary MCP-1 and IL-6 levels were increased significantly, while EGF excretion reduced in IgAN patients, compared to controls. IL-6 urinary levels showed significant positive correlation with chronic histological lesions. Patients were classified into five groups, according to the Haas classification system. MCP-1 and IL-6 urinary levels were increased, whereas EGF levels were reduced in the progression of staging. EGF urinary excretion was a strong predictor factor of disease outcome, significantly correlated with creatinine clearance at time of diagnosis ( r  = 0.5, P  = 0.005), and at the end of follow up ( r  = 0.6, P  = 0.001). Urinary EGF levels measured a year later could predict long-term outcome better, and a cut of 0.05 pg/mg urine creatinine levels could distinguish between progressors and non-progressors.
Conclusion:   Urinary MCP-1, IL-6 and EGF levels may represent histology in IgAN. EGF excretion can be a predictive marker and its serial measurements may give information about disease outcome and the effect of treatment.     

Monoclonal antibody 1-22-3-induced glomerulonephritis in uninephrectomized rats as a model of progressive renal failure.

BACKGROUND: At present, there are few available animal models of progressive renal failure originating from mesangial proliferative glomerulonephritis (GN). In the current study, we examined the usefulness of anti-Thy-1 monoclonal antibody (mAb) 1-22-3-induced GN in uninephrectomized rats as a model of progressive renal failure by analysing the similarities to human disease. METHODS: GN was induced by intravenous injection of mAb 1-22-3 into uninephrectomized male Wistar rats. The natural course of the disease was analysed in this model for 47 weeks. The effect of treatment with the angiotensin-converting enzyme inhibitor, captopril, on renal functional outcome was also examined in this model for 23 weeks, beginning from 1 week after antibody injection. RESULTS: Injection of mAb 1-22-3 induced a persistent proteinuria during the entire study period. Animals showed a progressive decline in renal function and 63% died by week 47. Severe glomerular and tubulointerstitial lesions were consistently observed. Treatment with captopril significantly inhibited increases in proteinuria and blood pressure, and attenuated renal injury. Captopril also retarded the progression of renal failure, and decreased mortality. Finally, the level of proteinuria was significantly correlated with the rate of decline in renal function, and the reduction in proteinuria by captopril was accompanied by a slower progression of renal failure. CONCLUSIONS: The mAb 1-22-3-induced GN in a uninephrectomized rat model simulates the clinical manifestations of human disease, indicating that this model may be useful for studying progressive renal failure and for investigating new therapeutic strategies against renal failure.