Antinephritic effects of PGE1 and thiaprostaglandin E1, TEI-5178 and TEI-6122, on crescentic-type anti-GBM nephritis in rats

The antinephritic effects of PGE1, TEI-5178 and TEI-6122 on crescentic-type anti-glomerular basement membrane (GBM) nephritis in rats were investigated. The test compounds were subcutaneously administered every day for 39 days after the injection of anti-GBM serum. PGE1 (2.0 mg/kg/day), TEI-5178 (0.25 or 0.5 mg/kg/day) and TEI-6122 (0.25 or 0.5 mg/kg/day) significantly reduced urinary protein by 30 to 50% of that of the control at the late stage of nephritis. These test compounds also suppressed the increase of blood urea nitrogen and the development of alteration in the glomeruli by the 40th day. Both TEI-5178 (0.5 mg/kg/day) and TEI-6122 (0.5 mg/kg/day) significantly suppressed the production of antibody to rabbit gamma-globulin in nephritic rats. This was not the case with PGE1, however. In additional experiments to clarify the antinephritic mechanisms of the test compounds, it was found that 15 min after one subcutaneous injection of PGE1 (1.0 mg/kg), TEI-5178 (0.5 mg/kg) or TEI-6122 (0.5 mg/kg), systolic blood pressure in the nephritic rats was transiently reduced by 50 to 60%. On the other hand, these test compounds augmented renal blood flow (20-50%) from 45 min after the injection. The relationship between the antinephritic effect and these subsequent findings will be discussed.     

肿瘤相关巨噬细胞对非小细胞肺癌MCP-1、MIP-1的影响

目的 观察肿瘤相关巨噬细胞(TAMs)U937和非小细胞肺癌细胞(NSCLC)A549在体外不同的共培养条件下,对单核细胞趋化蛋白-1(MCP-1)、巨噬细胞炎性蛋白-1(MIP-1)表达的影响.方法 采用Jeremy培养方法,分为三组:1组用不同浓度的U937细胞与A549细胞共培养24h;2组同浓度的U937细胞与A549细胞共培养不同时间;3组用A549在不同时间段单独培养,采用原位杂交方法分别检测三组A549细胞中MCP-1、MIP-1mRNA表达情况.结果 随着U937细胞浓度逐渐增高以及共培养时间的逐渐延长,癌细胞A549中MCP-1、MIP-1mRNA表达的阳性系数均值也相应地增高,明显高于U937细胞浓度为0×105/ml的共培养组以及各时段对照组(均P<0.05).结论 TAMs细胞U937和NSCLC细胞A549的相互作用,与A549细胞MCP-1,MIP-1mRNA的阳性表达密切相关,并存在着浓度和时间的依从性.     

西替利嗪对皮肤细胞炎症中单核趋化蛋白-1表达的干预作用

目的考察西替利嗪(CET)对皮肤细胞炎症模型中单核趋化蛋白-1(MCP-1)表达的干预作用。方法组胺(HIS)和IFN-γ刺激真皮成纤维细胞(DF)和人角质形成细胞株HaCaT细胞，采用RT-PCR和ELISA法考察两种细胞MCP-1 mRNA和蛋白表达水平。结果与对照组比较，HIS(10 μmol·L^-1)和IFN-γ(20 ng·mL^-1)组显著上调两种细胞(DF和HaCaT) MCP-1 mRNA表达，同时分别使DF的MCP-1蛋白分泌量增加3.5倍和8.4倍，对HaCaT细胞也有相似的影响趋势； CET (1和10 μmol·L^-1) 显著地抑制了它们对细胞MCP-1蛋白产生的增强作用。结论CET可能通过抑制MCP-1的表达而发挥抗皮肤炎症作用。     

Aspirin and PPAR-alpha activators inhibit monocyte chemoattractant protein-1 expression induced by high glucose concentration in human endothelial cells

Activated endothelial cells express monocyte chemoattractant protein-1 (MCP-1), a chemokine which is reportedly involved in the recruitment of plasma monocytes in the early stages of atherosclerosis. Since accelerated atherosclerosis is the main complication of diabetes and both diseases encompass an inflammatory reaction, we hypothesized that the anti-inflammatory drugs, aspirin and peroxisome proliferator-activated receptor (PPAR-alpha) activators (fenofibrate and clofibrate), could have an effect on the high glucose-induced MCP-1 expression in endothelial cells. To test this assumption, as well as the possible mechanisms involved, the MCP-1 expression and secretion, the reactive oxygen species levels, nuclear factor-kB (NF-kB) and activator protein-1 (AP-1) expression were determined in human endothelial cells exposed to high glucose concentrations in the presence of aspirin, fenofibrate and clofibrate. Human endothelial cells kept in normal glucose concentration in the absence of drugs were used as control. The results showed that (i) aspirin, fenofibrate and clofibrate decrease significantly the MCP-1 expression and secretion in human endothelial cells; (ii) the high glucose up-regulated expression of MCP-1 in endothelial cells was significantly reduced by inhibitors of NF-kB and reactive oxygen species; (iii) all drugs notably decrease the level of the reactive oxygen species and activation of NF-kB and AP-1. Together, the findings indicate that in endothelial cells aspirin and PPAR-alpha activators reduce the high glucose-increased expression of MCP-1 by a mechanism that includes the inhibition of reactive oxygen species, and decrease of AP-1 and NF-kB activation.     

MCP-1在IgA肾病中的临床意义

目的 探讨单核细胞趋化蛋白1(MCP-1)在IgA肾病中的变化及其临床意义.方法 用ELISA法检测38例lgA肾病患者和20名健康对照者MCP-l,并比较其差异;测定IgA肾病患者血及尿β2微球蛋白(β2MG)、24-h尿蛋白定量、血清肌酐(SCr);检查肾脏病理损伤,评价MCP-1与肾脏病理损伤的相关性,及在不同病理类型之间的差异.结果 与对照组相比,IgA肾病患者尿MCP-1显著升高(120.90 pg/ml vs.46.82 pg/ml),而血MCP-1差异无统计学意义.IgA肾病中,尿MCD-1与肾小管间质病变程度、24-h尿蛋白、尿β2MG呈正相关,尿MCP-1水平在轻微病变患者中明显低于局灶节段增生型、系膜增生型、硬化型患者.结论 尿MCP-1可作为评价IgA肾病患者肾小管间质功能的一项无刨伤性指标.     

Induction of heme oxygenase-1 inhibits monocyte chemoattractant protein-1 mRNA expression in U937 cells

Heme oxygenase-1 (HO-1) is a stress-inducible isoform of HO with potential cytoprotective effects. Monocyte activation/migration mediated by monocyte chemoattractant protein-1 (MCP-1) is one of the earliest and important events in the pathogenesis of atherosclerosis. We examined the effect of HO-1 on the production of lysophosphatidylcholine (Lyso-PC)-induced MCP-1 in the human promonocytic cell line U937. Increased HO-1 induction by hemin resulted in a significant decrease in the Lyso-PC-mediated induction of MCP-1 mRNA expression. SnPP (IX), the specific inhibitor of HO-1 enzymatic activity, prevented the hemin-mediated attenuation of MCP-1 mRNA expression. These results suggest that HO-1 may work as an anti-atherogenic agent through the attenuation of MCP-1 production.     

Intracellular redox status modulates monocyte chemoattractant protein-1 expression stimulated by homocysteine in endothelial cells

Homocystinemia has been identified as an independent risk factor for atherosclerosis. Monocyte chemoattractant protein-l (MCP-l) is a potent chemokine that stimulates the migration of monocytes into the intima of the arterial wall. The authors investigated the role of intracellular redox status in the expression of MCP-l stimulated by homocysteine in endothelial cells. Homocysteine stimulated MCP-1 mRNA expression and protein production in a time-dependent and dose-dependent manner in endothelial cells, decreased intracellular glutathione (GSH) and protein thiol levels, as well as G6PDH activity and NADPH levels. Thiol reduced reagents, GSH, and dithiothreitol levels, and reversed the MCP-l mRNA expression and protein production in endothelial cells; in addition, thiol oxidized reagent, diamide, and BSO levels, and markedly potentiated homocysteine-mediated up-regulation of MCP-l mRNA expression and protein production in endothelial cells. These results demonstrate that homocysteine can trigger overexpression of the MCP-1 gene by altering the intracellular redox status, suggesting that the homocysteine-induced changes in the intracellular redox status play an important role in modulating the expression of MCP-l in endothelial cells.     

乙酰吗啡成瘾者外周血单个核细胞中单核细胞趋化蛋白-1 mRNA表达的抑制(英文)

目的:探究乙酰吗啡(海洛因)成瘾者单核细胞趋化蛋白-1(MCP-1)基因表达状况是否与正常人有区别.方法:以β-actin为内标准物,用逆转录聚合酶链式反应(RT-PCR)分别检测乙酰吗啡成瘾者和正常志愿者外周血单个核细胞中MCP-1 mRNA表达状况,并对RT-PCR产物进行测序,以证实其特异性.结果:外周血单个核细胞中MCP-1 mRNA表达的相对水平,正常志愿者对照组为0.47±0.12(n=15);乙酰吗啡成瘾组为0.21±0.09(n=21),两组间差异显著(P<0.05).结论:乙酰吗啡成瘾者外周血单个核细胞中MCP-1 mRNA的表达显著被抑制;这可能是乙酰吗啡成瘾者群体中包括AIDS等严重感染性疾病发病率显著增高的机制之一.     

二甲双胍降低2型糖尿病患者尿单核细胞趋化蛋白-1的排泄

目的 观察二甲双胍对2型糖尿病(DM)患者尿单核细胞趋化蛋白-1(MCP-1)排泄的影响,探讨二甲双胍对2型糖尿病的保护作用.方法 对50例初诊的2型糖尿病患者给予二甲双胍治疗12周,观察12周前后相关代谢指标及尿MCP-1排泄的变化,同期选择30例体检健康人群作对照分析.结果 (1)2型糖尿病患者空腹血糖,餐后2 h...     

单核细胞趋化蛋白-1在肾毒血清肾炎大鼠肾组织中的表达

目的：探讨单核细胞趋化蛋白－（ＭＣＰ－１）在肾毒血清肾炎大鼠肾组织中的表达及其意义。方法肾毒血清肾炎应用兔抗鼠肾小球基底膜肾毒血清制备。采用免疫组化及真彩色计算机图像分析系统观察肾小球及肾小管中ＭＣＰ－１的表达,并分析其与蛋白尿和肾小球细胞数之间的关系。结果肾毒血清肾炎大鼠出现大量蛋白尿,肾小球细胞数明显增多;肾小球及肾小管中ＭＰＣ－１表达显上调,并与肾小球细胞数和蛋白尿密切相关。结论ＭＣＰ－１     

Oxidative stress mediates sodium arsenite-induced expression of heme oxygenase-1, monocyte chemoattractant protein-1, and interleukin-6 in vascular smooth muscle cells.

Arsenic exposure is associated with an increased risk of vascular disorders, and results in increased oxidative stress in endothelial cells and vascular smooth muscle cells (VSMCs). Since oxidative stress is involved in regulating the expression of genes related to atherogenesis, we investigated its involvement in the enhanced expression of three atherosclerosis-related genes coding for heme oxygenase-1 (HO-1), monocyte chemoattractant protein-1 (MCP-1), and interleukin-6 (IL-6) in VSMCs treated with inorganic sodium arsenite (iAs). In human VSMCs (hVSMCs) and rat VSMCs (rVSMCs), HO-1, MCP-1, and IL-6 mRNA levels were significantly increased by iAs treatment. An increase in HO-1 protein levels in hVSMCs was confirmed by Western blotting technique, while increased MCP-1 and IL-6 secretion by hVSMCs was demonstrated by enzyme-linked immunosorbent assay. Although modulators of oxidative stress inhibited this iAs-induced increase in the expression of these three genes, different modulators had differential effects. In iAs-treated rVSMCs, catalase, dimethylsulfoxide, and L-omega-nitro-L-arginine significantly inhibited the increase in expression of all three genes, allopurinol inhibited the increase in MCP-1 and IL-6 expression, but had no effect on HO-1 expression, while superoxide dismutase had no significant effect on HO-1 expression, but had an inhibitory effect on IL-6 expression and a stimulatory effect on MCP-1 expression. Therefore, iAs may enhance the expression of HO-1, MCP-1, and IL-6 in VSMCs via different reactive oxygen molecules. Furthermore, using tin protoporphyrin IX (SnPP) and anti-MCP-1 antibody to abolish iAs-induced HO-1 and MCP-1 activity, respectively, shows that HO-1 has protective effect against iAs-induced injury in VSMCs and MCP-1 is chemoattractive to human monocytes, THP-1.     

Effects of atorvastatin,simvastatin, and fenofibrate therapy on monocyte chemoattractant protein-1 secretion in patients with hyperlipidemia

OBJECTIVE: Monocytes that migrate into the arterial wall participate in the development and, eventually, rupture of the atherosclerotic plaque. The aim of this study was to evaluate the secretion of monocyte chemoattractant protein-1 (MCP-1) by monocytes from hyperlipidemic patients treated with hypolipidemic drugs, namely fenofibrate, simvastatin, or atorvastatin to determine what role is played by these drugs in the development and stabilization of the atherosclerotic plaque. METHODS: Fifty-four hyperlipidemic patients, who did not respond to a low-fat diet, were treated with fenofibrate, simvastatin, or atorvastatin (18 patients in each group) for 1 month. The control group included 18 normolipidemic, healthy, age-matched participants. Ten hyperlipidemic patients were effectively treated with hypolipidemic diet alone for 1 month. This group was compared with a control group of ten healthy subjects. To accurately evaluate the adhesion molecule levels, we excluded hyperlipidemic patients and control subjects with any inflammatory disease. Before and after treatment, monocytes were isolated from peripheral blood. After stimulation with lipopolysaccharide (LPS), MCP-1 secretion was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: MCP-1 levels were significantly higher in hyperlipidemic patients than controls: 15.8+/-0.47, 16.7+/-0.23, and 14.9+/-0.45 compared with 12.36+/-0.42 ng/ml. Fenofibrate, atorvastatin, and simvastatin significantly decreased MCP-1 levels from 15.8+/-0.47 to 8.79+/-0.89, from 16.7+/-0.23 to 7.46+/-0.73, and from 14.9+/-0.45 to 10.3+/-0.8 ng/ml, respectively. In the diet-treated group of hyperlipidemic patients, the level of MCP-1 before therapy was significantly higher than in controls (16.89+/-0.31 vs 12.45+/-0.36 ng/ml). The diet therapy caused a significant decrease in levels of MCP-1 to 15.1+/-0.36 ng/ml. There was a correlation between the decreased levels of lipids and the decreased release of MCP-1 in the patients treated with hypolipemic drugs. CONCLUSION: The drug-induced decrease in MCP-1 secretion in hyperlipidemic patients suggests that, apart from acting on lipids, the hypolipidemic drugs studied may directly inhibit the activity of monocytes.     

油酰乙醇胺对脂多糖诱导的THP-1细胞炎症因子表达的影响

目的探讨油酰乙醇胺(OEA)对细菌脂多糖(LPS)诱导的人急性白血病单核细胞(THP-1)中前炎症因子TNF-α、IL-1β、IL-6表达的影响,并初步探讨OEA作为过氧化物酶体增殖物激活受体-α(PPAR-α)激动剂参与对炎症调节的作用机制。方法体外培养的THP-1细胞,分别加入不同浓度的OEA(10,20,40μmol/L)或非诺贝特(100μmol/L)共同孵育1 h后,用1μg/mL LPS分别诱导6或24 h。采用RT-PCR、实时定量PCR和酶联免疫吸附检测测定细胞中TNF-α、IL-1β、IL-6 mRNA和蛋白的表达的变化,并使用实时定量PCR及Western blot方法检测PPAR-α及Toll样受体4(TLR4)的mRNA和蛋白的表达。结果相对于正常THP-1细胞,LPS诱导后细胞中炎症因子(TNF-α、IL-1β、IL-6)表达明显增加。OEA对TNF-α、IL-1β、IL-6 mRNA和蛋白的表达有抑制作用,并呈现出一定的剂量依赖性。且OEA在激活PPAR-α表达的同时能够抑制TLR4的表达。结论 OEA对LPS诱导的炎症反应有抑制作用,其机制可能与激活PPAR-α,下调TLR4的表达有关。     

Change of concept about the regulation of angiotensin II-induced monocyte chemoattractant protein-1 production in human endothelial cells

AimsSome intriguing clinical observations about the anti-inflammatory effects of angiotensin type 1 (AT1) receptor blockers and angiotensin converting enzyme inhibitors in cardiovascular patients brought us to study the signalling pathways which lead to angiotensin II (ANG)-induced monocyte chemoattractant protein-1 (MCP-1) production in human endothelial cells.MethodsMCP-1 production in human umbilical vein endothelial cells (HUVECs) under treatments with ANG, AT1 and angiotensin type 2 (AT2) receptor blockers and pravastatin was measured by ELISA. The expression of AT1 and AT2 receptors and NADPH oxidase catalytic subunits (NOX 1–5) was analysed at mRNA and protein levels. Nuclear factor-kappa B (NF-κB) activation was studied by p65 subunit translocation to the cellular nucleus. Cell viability was tested by the MTT method. Nox4 subcellular distribution was analysed by subcellular protein fractionation and by immunoprecipitation followed by matrix-assisted laser desorption/ionization mass spectrometry analysis.ResultsANG-induced MCP-1 production was mediated by AT2 receptor, but not AT1 receptor in HUVECs in culture, which in turn activated NF-κB, promoting p65 subunit translocation to the nucleus. Reactive oxygen species produced by NADPH oxidase participated in this activation, mainly by the Nox4 subunit, ubiquitously expressed in all the compartments of HUVECs. Pravastatin inhibited ANG-induced MCP-1 production.ConclusionsOur results support that ANG-induced MCP-1 production in HUVECs is mediated by AT2 instead AT1 receptor activation, which in turn activates NF-κB involving reactive oxygen species produced by the NADPH oxidase complex. Statins can also block ANG-induced MCP-1 production, probably by their inhibitory effects on NADPH oxidase activity.     

Effect of monocyte chemoattractant protein-1 on murine bone marrow cells: proliferation,colony-forming ability and signal transduction pathway involved

Monocyte chemoattractant protein-1 (MCP-1) plays a crucial role in the migration and activation of leukocytes in both physiological and pathological contexts. In this paper, we report the in vitro effect of MCP-1 on myeloid haematopoiesis. MCP-1-treated murine nonadherent bone marrow cells (NABMCs) were assayed for in vitro proliferation and colony forming ability. It is observed that MCP-1 treatment in vitro caused an enhancement in the proliferation and colony forming ability of the murine NABMCs as compared to the untreated cells. This response was concentration-dependent and most effective at a dose of 100ng/ml MCP-1. In the presence of MCSF (200U/ml), GCSF (200U/ml), GMCSF (200U/ml) or IL-3 (200U/ml), the MCP-1-induced colony forming ability of the NABMCs was significantly augmented, indicating a synergistic effect of MCP-1 with these CSFs. However, irrespective of the CSFs used, MCP-1 stimulated the lineage-restricted differentiation of the murine BMCs into predominantly the granulocytic lineage. NABMCs cultured in medium alone formed minimal colonies. The probable signal transduction mechanism responsible for the MCP-1-induced NABMC proliferation/differentiation was also investigated. The results of the colony forming assay indicate that the protein kinase inhibitors, genistein (10&mgr;g/ml), chelenthryin chloride (10&mgr;M), wortmannin (200nM) and PD98059 (10&mgr;M) significantly blocked the in vitro colony forming ability of the MCP-1-treated NABMCs, while the phosphatase inhibitors, okadaic acid (10nM) and sodium orthovanadate (10&mgr;M) caused an increase in the BMC colony forming ability in response to MCP-1. These data suggests the involvement of the respective protein kinases and phosphatases in the above process. Correlating with this, the role of several signaling molecules likes Lyn, p42/44MAPK, PI3K and STAT5 has also been implicated in the signal cascade of murine NABMC proliferation/differentiation following MCP-1 treatment.     

血管紧张素Ⅱ及其受体拮抗剂对人肾小管细胞MCP-1mRNA表达的影响

近年来研究表明,肾脏局部肾素血管紧张素系统(RAS)的激活在慢性肾病进展中起着十分重要的作用,肾小管上皮细胞在促进小管间质炎症反应和纤维化形成中起十分关键的作用[1,2].单核细胞趋化蛋白-1(MCP-1)是趋化因子家族的重要一员,在肾脏的炎症反应中对炎症细胞的趋化和激活有重要影响.文献中曾报道血管紧张素Ⅱ(AngⅡ)可以诱导系膜细胞MCP-1 mRNA表达[3],但AngⅡ对小管上皮细胞MCP-1表达影响如何,文献报道尚少.因此,本实验通过体外培养的人近端肾小管细胞(HPTC),探讨AngⅡ对HPTC MCP-1 mRNA表达的影响,以进一步阐明AngⅡ、MCP-1在介导肾间质纤维化中的作用机制,为临床合理应用RAS阻断剂治疗慢性肾病提供理论依据.     

地塞米松对实验性自身免疫性脑脊髓炎大鼠MCP-1表达的影响

目的为探讨地塞米松(DXM)对实验性自身免疫性脑脊髓炎(EAE)大鼠临床指标和单核细胞趋化蛋白1(MCP1)mRNA的影响。方法将正常大鼠、完全抗原和百日咳毒素原液免疫后的大鼠免疫后7d,分成:正常大鼠、EAE组和DXM组。DXM组给予DXM腹腔注射,另外两组给予生理盐水注射。取脑和脊髓制成石蜡切片,进行苏木素伊红(HE)染色和MCP1mRNA的原位杂交(ISH),并比较各项临床指标。结果DXM组的发病率、临床评分和MCP1mRNA的阳性细胞数显著性降低(P<0.01)。结论进一步证实DXM的使用可对EAE有治疗作用。