单核细胞趋化蛋白-1与糖尿病肾病

单核细胞趋化蛋白-1(MCP-1)是一种特异性趋化因子，在糖尿病肾病 (diabetic nephropathy，DN)的发生发展中起重要作用。代谢性因素如高糖（HG）、糖基化白蛋白（glycated albumin， Gly-Alb）、氧化应激，蛋白激酶C(protein kinase C，PKC)等和血流动力学因素如肾素-血管紧张素系统（RAS）等均可上调肾小球内皮细胞、系膜细胞 (mesangial cell，MC)、小管上皮细胞（tubulus epithelial cells，TECs）中MCP-1基因和蛋白的表达，使单核/巨噬细胞在肾组织中聚集，通过多种机制引起肾脏损伤。     

高浓度的糖刺激人肾小球内皮细胞表达单核细胞趋化蛋白-1的影响

目的 :探讨高浓度的糖对培养的人肾小球内皮细胞 (HUGEC)表达单核细胞趋化蛋白 1(MCP 1)的影响 ,以及HUGEC的条件培养基对单核细胞 (MC)的趋化作用及抗MCP 1抗体对MC迁移的影响。方法 :采用原位杂交技术和细胞ELISA法 ,观察MCP 1的表达 ;用改良的Boyden小室微孔滤膜法 ,测定高浓度的糖刺激HUGEC后的条件培养基 ,对MC的趋化作用 ,以及抗MCP 1抗体对MC迁移的影响。结果 :(1)在低浓度的糖(含 5 .5mmol/LD 葡萄糖 )条件下培养的HUGECMCP 1mRNA呈弱表达。高浓度的糖 (含 2 5mmol/LD 葡萄糖 )刺激后 ,8h即出现MCP 1表达增强 ,于 16h达高峰。(2 )高浓度的糖刺激HUGEC后的条件培养基 ,对MC具有明显的趋化作用 ;抗MCP 1抗体可抑制其作用。结论 :在高浓度的糖诱导下 ,人HUGECMCP 1的表达增强 ,其条件培养基对MC具有趋化作用 ,从而可能招引单核细胞迁入内皮下间隙。     

睾酮双向影响脐静脉内皮细胞分泌单核细胞趋化蛋白1

雄激素与动脉硬化(arteriosc lerosis,AS)的关系日益受到关注,单核细胞趋化蛋白1(monocyte chemoattractant prote in-1,MCP-1)是促进AS的重要炎症介质。本研究以人脐静脉内皮细胞(hum an umb ilical ve in endothelial cell,HUVEC)为对象,观察不同浓度睾酮对MCP-1蛋白表达的影     

The effect of inflammatory cytokines on secretion of macrophage colony-stimulating factor and monocyte chemoattractant protein-1 in human granulosa cells

PROBLEM: In order to investigate the role of macrophage colony-stimulating factor (M-CSF) and monocyte chemoattractant protein -1 (MCP-1) in human ovulation, we studied the regulation of M-CSF and MCP-1 in cultured human granulosa cells. METHOD OF STUDY: Immortalized granulosa cells (GC1a) were cultured in serum-free medium, and incubated with interleukin (IL)-1alpha, IL-1 receptor antagonist (ra) and tumor necrosis factor (TNF)-alpha. The supernatants were collected, and M-CSF and MCP-1 were measured by ELISA. RESULTS: The levels of M-CSF and MCP-1 were increased after treatment with IL-1alpha (1 nm) and TNF-alpha (1 nm) in a time-dependent manner. The levels of M-CSF and MCP-1 were significantly increased after treatment with IL-1alpha and TNF-alpha in a dose-dependent manner. However, the levels of M-CSF and MCP-1 were significantly decreased by treatment with IL-1alpha (1 nm) and/or increasing concentrations of IL-1 ra. CONCLUSIONS: Our data indicated that M-CSF and MCP-1 were regulated by IL-1alpha and TNF-alpha. It was suggested that M-CSF and MCP-1 may play an important role in human preovulatory processes.     

Cysteinyl leukotrienes induce monocyte chemoattractant protein 1 in human monocytes/macrophages

BACKGROUND: Monocytes/macrophages have a cysteinyl leukotriene 1 (CysLT1) receptor, but its function is poorly understood. Objective To elucidate the biological function of the CysLT1 receptor of human monocytes/macrophages. METHODS: We examined the production of TNF-alpha, IL-1beta, IL-2, IL-4, IL-6, IL-8, IL-10, monocyte chemoattractant protein 1 (MCP-1), macrophage colony-stimulating factor (M-CSF), and eotaxin induced by CysLTs (leukotriene (LT)C4, -D4, and -E4) in THP-1 cells, a human monocytic leukaemia cell line, and peripheral blood CD14+ monocytes/macrophages. Moreover, we examined the effect of CysLTs on the expression of beta-chemokine receptor 2B (CCR2B) as the receptor of MCP-1 by Western blot analysis. RESULTS: ELISA revealed that CysLTs induced MCP-1 in THP-1 cells and peripheral blood CD14+ monocytes/macrophages, but not other cytokines. PCR demonstrated that CysLTs increased MCP-1 mRNA expression in THP-1 cells, and Western blotting showed that CysLTs increased the expression of CCR2B in THP-1 cells. Moreover, we demonstrated that pranlukast, a CysLT1 receptor antagonist, blocked MCP-1 production by CysLTs in THP-1 cells almost completely, and partially inhibited MCP-1 release by CysLTs in peripheral blood CD14+ monocytes/macrophages and CCR2B expression by CysLTs in THP-1 cells. CONCLUSION: CysLTs induce MCP-1 and increase CCR2B expression in human monocytes/macrophages.     

Deficiency in monocyte chemoattractant protein-1 modifies lipid and glucose metabolism

We describe the effect of MCP-1 deficiency in mice rendered hyperlipemic by the concomitant ablation of the LDL receptor. The MCP-1(-/-)LDLr(-/-) mice in comparison with LDLr(-/-) mice showed a decreased lipoprotein clearance, derangements in free fatty acids delivery and less glucose tolerance when fed a regular chow, and they showed a partial resistance to alterations in glucose and lipid metabolism induced by dietary fat and cholesterol. They also were less prone to the development of diet-induced obesity. Our results suggest that the role of MCP-1 in metabolism is relevant and that, although new hidden complexities are evident, the function of MCP-1/CCL2 extends far beyond the monocyte chemoattractant effect. Therefore, the regulatory mechanisms influenced by MCP-1 should be fully ascertained to understand the metabolic consequences of inflammation and before considering MCP-1 as a therapeutic target.     

实验性变态反应性脑脊髓炎大鼠单核细胞趋化蛋白-1的表达与临床评分的相关性

目的：探讨实验性变态反应性脑脊髓炎（EAE）大鼠脑和脊髓中单核细胞趋化蛋白-1（MCP-1）的表达及其与临床评分的关系。方法：将Wistar大鼠分为正常组、佐剂（CFA）组和EAE组,取脑和脊髓制成石蜡切片,进行HE染色和MCP-1mRNA的原位杂交,并与各项临床指标比较。结果：EAE组的体重减轻、MCP-1 mRNA表达的阳性细胞百分数与正常大鼠、佐剂组相比显著性增加。EAE组大鼠MCP-1 mRNA的表达呈动态性变化,其MCP-1 mRNA先于临床症状高表达,并随临床评分的升高而升高,与临床评分呈正相关。结论：MCP-1是参与EAE发病的重要的炎性介质。     

硒对糖尿病患者的内皮细胞表达P38信号通路和MCP-1的影响

目的:探讨P38MAPK和MCP-1的关系及其在糖尿病动脉粥样硬化中的作用以及硒的作用机制。方法:分别以高葡萄糖、糖基化终末产物(AGE)、胰岛素或过氧化氢体外孵育人脐静脉内皮细胞(HUVEC),并以Westernblot法或RT-PCR法检测P38MAPK和MCP-1在HUVEC的表达。检测并比较有无P38MAPK特异抑制剂SB203580或硒预处理,以上4种因素对P38MAPK和MCP-1在HUVEC表达的影响。结果:高葡萄糖、AGE、胰岛素或过氧化氢均可独立激活P38MAPK,并增加MCP-1在HUVEC的表达。SB203580显著抑制MCP-1的表达。硒抑制P38MAPK的活化并减少MCP-1的表达。结论:P38MAPK是MCP-1的上游信号分子,表明P38MAPK可能是糖尿病动脉粥样硬化发生的始动信号之一。硒可能通过抑制P38MAPK磷酸化而抑制MCP-1表达,有预防动脉粥样硬化作用。     

两步法细菌内同源重组制备重组单核细胞趋化因子-1腺病毒

目的两步法提高细菌内同源重组效率制备重组人单核细胞趋化蛋白1（MCP-1）复制缺陷型腺病毒。方法制备含有腺病毒骨架质粒pAdEasy—1的大肠杆菌BJ5183，HindⅢ酶切筛选未发生细菌内重组的菌落，并将其制备成感受态（BJ5183pAdEasy—1）RT—PCR法克隆得纠MCP—1 cDNA片段，连接到pMD18-T载体后亚克隆至腺病毒穿梭载体pAdTrack上陶建重组穿俊载体pAdTrackMCP—1.PmeⅠ酶切线性化并碱性磷酸酶处理pAdTrack—MCP—1，然后电穿孔转化感受态BJ5183pAdEasy-1。PacⅠ酶切筛选得到正确重组的腺病毒载体pad—MCP—1，将pAd—MCP—1转染入HEK293细胞中包装病毒颗粒，以PCR法鉴定？结果成功构建pad—MCP-1，效率町达50％以上，pad—MCP—1能有效转染HEK293细胞并在细胞内包装PCR鉴定病毒携带有MCP—1基因片段、结论两步法细菌内同源重组构建再组腺病毒载体较常规电穿孔共转化效率高．成功制备重组腺病毒颗粒为进一步研究MCP-1的作用提供了重要的方法？     

甘草甜素抑制肺纤维化大鼠肺组织单核细胞趋化蛋白-1的表达

目的: 观察甘草甜素对博来霉素(BLM)诱导的大鼠肺纤维化的干预作用及可能机制。方法: 随机将大鼠分为对照组、肺纤维化模型组、甘草甜素干预组。气管内注入博莱霉素造成动物模型后,于当天开始每天给药,分别于7 、28 d处死,取肺组织,行嗜伊红染色、Masson染色；检测肺组织匀浆中羟脯氨酸(HYP) 的变化; RT-PCR法测肺组织单核细胞趋化因子-1 mRNA的表达,免疫组化测肺组织单核细胞趋化因子-1蛋白的表达。结果: 干预组肺纤维化程度轻于模型组; 肺组织匀浆羟脯氨酸(HYP)含量显著低于模型组( P<0.01) ;在模型组 MCP-1第7 d表达就明显升高，第28 d下降，但仍高于对照组，干预组与模型组有同一规律，但均减弱，第7、28 d与模型组(M组)比较，P<0.05。结论: 甘草甜素能减轻博莱霉素诱导的大鼠肺纤维化，这种作用可能部分是通过抑制MCP-1的表达而实现。     

类风湿关节炎患者血清单核细胞趋化蛋白-1水平及与间质性肺疾病的关系

目的:研究单核细胞趋化蛋白-1(MCP-1)在类风湿关节炎(RA)发病中的作用,探讨其与RA继发间质性肺疾病(ILD)的关系及其可能的临床价值。方法:选择RA患者60例,按是否存在ILD,分为单纯RA组30例和RA继发ILD组30例,正常对照组20例,应用ELISA测定各组血清MCP-1水平,比较各组MCP-1水平的差异,同时比较各组中实验室指标IgG、IgA、IgM、α2-球蛋白、γ-球蛋白、RF、ESR、CRP及关节肿痛数的差异,并分析以上指标与血清MCP-1表达的相关性。结果:RA患者MCP-1的表达明显高于健康对照组,RA继发ILD组显著高于单纯RA组。RA患者血清MCP-1与IgG、IgM、γ-球蛋白均呈正相关。结论:MCP-1是RA发病中重要的细胞因子,在RA继发ILD过程中可能起着一定的促进作用。     

Expression of monocyte chemoattractant protein-1 and interleukin-8 receptors on subsets of T cells: correlation with transendothelial chemotactic potential

The differential expression of chemokine receptors may be an important mechanism for the regulation of T cell migration. To test this, we examined the expression and function of the monocyte chemoattractant protein (MCP)-1 and interleukin (IL)-8 receptors on various populations of T cells. Using a simple and reliable transendothelial chemotaxis assay, both MCP-1 and IL-8 were shown to be chemotactic for subsets of blood T cells, although the relative response varied from donor to donor. To examine receptor expression and correlate it with chemotaxis of T cell subsets, monoclonal antibodies (mAb) to the receptors were produced by immunizing mice either with synthetic peptides (MCP-1 receptor), or with receptor transfectants (IL-8 receptors A and B). A flow cytometric analysis of blood T cells with an anti-MCP-1 receptor mAb revealed low expression on the CD26^hi subset and undetectable expression on other T cells. Staining of T cells with anti-IL-8RA and anti-IL-8RB showed much higher levels of expression, but only on a subset of CD3⁺ cells which were CD8⁺ and CD56^?. That IL-8 and MCP-1 attracted distinct subsets of T cells was best illustrated using the CD26 marker, since IL-8R⁺ T cells were CD26^?, whereas T cells expressing detectable MCP-1R or which responded to MCP-1 in chemotaxis assays were CD26^hi. T cells activated in vitro with anti-CD3 up-regulated expression of the MCP-1 receptor, but not the IL-8 receptors, and were attracted to MCP-1 much more efficiently than resting T cells. These results show that there is a clear distinction between the IL-8- and MCP-1-responsive T cell populations and that chemokine receptor expression on T cells may be regulated with respect to lineage as well as cellular activation.     

MCP-1在妊娠期高血压疾病患者血清中的变化及意义

目的探讨妊娠期高血压疾病患者血清单核细胞趋化蛋白-1(MCP-1)变化的意义。方法用ELISA法对21例妊娠期高血压疾病患者(其中轻度子痫前期12例,重度子痫前期9例)血清MCP-1浓度进行测定,并选择同期30例正常孕妇作为对照组。结果妊娠期高血压疾病患者血清中MCP-1含量显著高于对照组孕妇,并随病情加重呈增加趋势。结论 MCP-1所参与的免疫反应可能是妊娠期高血压疾病的发病机制之一。     

MCP-1对培养的人肾小球内皮细胞表达ICAM-1的影响

目的研究单核细胞趋化蛋白 - 1(MCP- 1)对培养的人肾小球内皮细胞 (HU GEC)表达细胞间粘附分子 - 1(ICAM- 1)的影响。方法采用细胞 EL ISA法。结果 1培养的 HU GEC表面有少量 ICAM- 1表达 ,在 10 ng/ m L MCP- 1刺激后 ICAM- 1表达量增多 (P<0 .0 5 ) ,6 h即有 ICAM- 1表达增强 ,12 h达高峰 ,不同浓度的 MCP- 1(10、2 0、40 ng/ m L)刺激HU GEC18h后 ,ICAM- 1表达与对照组比较差异显著 (P<0 .0 1) ;2加入抗 MCP- 1抗体后 ,ICAM- 1表达量下降 ,与对照组比较无差异 (P>0 .0 5 )。结论 MCP- 1可刺激 HU GEC表达 ICAM- 1增加。     

核因子-κB/IκB信号通路介导实验性肾炎肾组织中单核细胞趋化蛋白-1表达

目的　探讨核因子 κB(NF κB) /IκB信号通路在肾毒血清性肾炎肾组织单核细胞趋化蛋白 1(MCP 1)表达中的作用。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。应用凝胶电泳迁移率 (EMSA)和Western印迹检测肾毒血清肾炎大鼠肾组织中NF κB活化、p6 5亚基核转位以及IκBα和IκBβ的降解 ;采用免疫组织化学及核酸酶保护法检测肾组织中MCP 1表达 ,并分析其与NF κB活化的关系。结果　模型组肾小球及肾小管中MCP 1表达分别为 (2 4 37± 7 0 6 )个 /肾小球横切面和 (5 4 78± 11 4 9) % ,较正常对照组显著升高 (P <0 0 1) ;肾组织中NF κB活化显著增强 ,p6 5由胞质转移至胞核 ,胞质内IκBα和IκBβ降解明显增加 ;NF κB活化与MCP 1表达呈显著正相关。 结论　NF κB/IκB信号通路介导肾小球肾炎肾组织中MCP 1表达。     

流体切应力下血管内皮单核细胞趋化蛋白-1的表达

目的研究流体切应力对人血管内皮细胞的单核细胞趋化蛋白-1(MCP-1)表达的影响,探讨血流动力在动脉粥样硬化(AS)发生早期中的作用.方法利用平行板流动腔对血管内皮细胞施加不同切应力,分别采用夹心酶联免疫吸附测定(EL,ISA)及逆转录-聚合酶链反应(RT-PCR)法检测MCP-1蛋白及mRNA水平.结果以0.72Pa切应力进行不同时间作用,0.5h后MCP-1mRNA即达到静态时(0.160±0.037)的4倍(0.684±0.033),5h时略有升高,达0.707±0.089,12h后下降到低于对照组的水平(0.036±0.006,P＜0.001).灌流液中MCP-1蛋白时间依赖性增加,但5h后增长趋缓;而在不同切应力(0.30、0.72、2.40Pa)相同时间(5h)下,0.72Pa的作用最显著,MCP-1蛋白比静态对照增加了2倍多,mRNA水平则升高达4倍.结论MCP-1的表达对切应力的变化反应强烈,持续稳定的切应力则下调MCP-1的表达,此研究结果表明血流紊乱可促进AS病变的发生发展.     

CD16+ monocytes in breast cancer patients: expanded by monocyte chemoattractant protein-1 and may be useful for early diagnosis

Human peripheral blood monocytes are a heterogeneous population, including CD14⁺CD16^‐‘classical’ monocytes and CD14⁺CD16⁺‘proinflammatory’ monocytes. CD16⁺ monocytes are expanded in various inflammatory conditions. However, little is known about the CD14⁺CD16⁺ monocytes in patients with breast cancer. We detected CD14⁺CD16⁺ monocytes in 96 patients with breast cancer and 54 control subjects using flow cytometry. Receiver‐operating characteristic (ROC) curve analysis was used to determine the feasibility of CD14⁺CD16⁺ monocytes as an indicator for diagnosis of breast cancer. We found that the frequency of CD14⁺CD16⁺ monocytes showed a significantly greater increase in breast cancer patients than in controls (16·96% versus 10·84%, P < 0·0001). The area under the ROC curve for CD14⁺CD16⁺ monocytes was 0·805 [95% confidence interval (95% CI): 0·714–0·877, P = 0·0001]. Furthermore, the levels of CD16⁺ monocytes were significantly negatively associated with the tumour size and pathological staging. In vitro, we showed that CD14⁺CD16⁺ monocytes were expanded significantly when the purified CD14⁺ monocytes were exposed to Michigan Cancer Foundation (MCF)‐7 cells‐conditioned medium (MCF‐CM) or, separately, to monocyte chemotactic protein 1 (MCP‐1). Neutralizing antibodies against MCP‐1 inhibited the expansion of CD14⁺CD16⁺ monocytes by MCF‐CM. Collectively, our findings indicated that MCP‐1 can expand CD14⁺CD16⁺ monocytes in patients with breast cancer. Furthermore, the CD14⁺CD16⁺ monocyte may be a useful indicator in early diagnosis of breast cancer.     

Potential role for monocyte chemotactic protein-4 (MCP-4) in monocyte/macrophage recruitment in acute renal inflammation

The CC chemokine, monocyte chemoattractant protein-4 (MCP-4), is an important chemoattractant for monocytes and T cells. Recent data indicate a role in renal inflammation. This study has used in situ hybridization and immunohistochemical analysis of cryostat sections of biopsy material taken from patients with acute renal allograft rejection and vasculitic glomerulonephritis to demonstrate renal expression of MCP-4, both at message and protein level. MCP-4 was primarily expressed at peritubular, periglomerular, and perivascular sites, irrespective of the inflammatory condition, and was associated with infiltrating CD3-positive lymphocytes and CD68-positive monocyte/macrophages. In addition, proximal tubular epithelial cells grown in culture from cortical fragments of human kidney showed low levels of constitutive MCP-4 expression, detectable by western blotting; this expression of MCP-4 was up-regulated in response to the pro-inflammatory cytokines, tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). CCR3-, CCR5- and CCR2-expressing leukocyte populations were identified at sites of MCP-4 expression. Double-staining techniques revealed that CC chemokine receptor-expressing cells were primarily CD68-positive. These studies suggest an important role for MCP-4 in the recruitment and retention of monocytes/macrophages in renal inflammation.