人单核细胞趋化因子蛋白—1的临床研究进展

ＭＣＰ－１趋化因子家族的一个主要成员，具有诱导单核细胞趋化及激活单核细胞的双重功能，在人体各器官事均有表达，同时又地机体的防御、炎症恢复及抗肿瘤发挥重要作用。本文就人单核细胞趋化蛋白－１（ＭＣＰ－１）近的来在临床方面的研究进展作一综述。     

脉冲电磁场对细胞作用的特异性研究

本文研究了脉冲电磁场（ＥＭＦ）对细胞作用的特异性，并初步探索了作用机理。     

人单核细胞趋化蛋白-1的研究进展

人单核细胞趋化蛋白-1(MCP-1)为一由76个氨基酸组成的单肽链,它既具有特异的单核细胞趋化活性,也具有激活单核细胞的活性,在机体防御、炎症恢复和抗肿瘤等方面起着重要作用。本文就MCP-1近几年来的研究进展作一综述。     

低频脉冲电磁场对冻伤组织血液循环的影响

目的:研究观察频率为15 Hz、强度150 mT占空比一定的低频脉冲电磁场(Pulsed electromagnetic fields,PEMFs)对SD大鼠冻伤部位组织血液循环的影响。方法:设计以高精度、低输出阻抗信号发生器芯片MAX038为核心的低频脉冲电磁场发生器,装置可产生频率为15 Hz,强度150 mT的电磁场;在大鼠的背部区域,采用液氮冷却的方法,建立中度冻伤的SD大鼠冻伤模型。将24只大鼠分为3组,空白对照组,单纯冻伤组,冻伤照射治疗组。采用该低频脉冲电磁场照射冻伤治疗组,6 h/d,持续10 d。然后,采用动物B超检查各组大鼠冻伤部位组织血液循环情况,并测量各组大鼠冻伤部位存活面积。结果:各组SD大鼠冻伤后存活面积测量显示照射组存活面积最大(P<0.05);对比各组大鼠冻伤部位B超图像,表明照射组血液循环明显优于单纯冻伤组。结论:一定强度的低频脉冲电磁场能有效改善冻伤部位组织血液循环,促进冻伤恢复。     

内源性接触激活系统对单核细胞粘附的影响

尿激酶型纤溶酶原激物（U-PA）是近年来被广泛研究的生物活性大分子，研究表明，它在单核细胞粘附中起着重要作用，由于U-PA 与纤溶酶原（Plasminogen）十二因子（Ⅻ）激肽释放酶（KK）及高分子激肽原（HK）在功能上密切相关，故本文在体外研究四种因子对U-PA 引起单核细胞粘附的影响。1 方法小鼠单核细胞系U937 细胞和3H-TdR 共同培养24h，离心洗涤后重悬细胞（1×10~6/mL），分别加入U-PA，纤溶酶原，Ⅻ，KK，HK，在24 孔板中（300 m L/孔）培养17h， 去上清，残留的细胞即为粘附在24 孔上的单核细胞。每孔加入1mL 裂解液，再…     

脉冲电磁场对细胞的特异性作用研究

本文研究了脉冲电磁场（ＥＭＰ）对细胞作用特异性，并初步探讨了作用机理。     

极低频脉冲电磁场对新生大鼠成骨细胞的影响

目的：观察极低频脉冲电磁场（pulsed electromagnetic fields，PEMF）对体外培养成骨细胞增殖、分化、体外矿化的影响。方法：采用频率为15Hz、强度为5mT、占空比为15％的PEMF作用于成骨细胞，检测成骨细胞的增殖、碱性磷酸酶（alkaline phosphatase，ALP）活性以及体外矿化指标。结果与结论：PEMF显著促进成骨细胞增殖和体外矿化，抑制ALP活性作用。     

低剂量内皮-单核细胞激活多肽II对人U251胶质瘤细胞迁移和侵袭的影响

目的研究低剂量内皮-单核细胞激活多肽II(EMAP II)对U251胶质瘤细胞迁移和侵袭行为的影响及相关分子机制。方法 EMAP II(0.05n M)分别处理U251细胞不同时间后,采用Transwell小室实验及Matrigel Transwell小室实验检测U251细胞迁移和侵袭的变化,应用Western blot方法检测Fox O1和p-Fox O1的表达变化;转染Fox O1的沉默质粒(sh Fox O1)到U251细胞中,检测Fox O1敲减后EMAP II作用下U251细胞迁移和侵袭行为的改变。结果 EMAP II作用0.5 h和1 h时显著抑制U251细胞的迁移和侵袭行为;与EMAP II作用0 h组相比,Fox O1的表达在EMAP II作用0.5 h和1 h时显著升高,而p-Fox O1的表达显著下降;此外,sh Fox O1可减弱EMAP II对U251细胞迁移和侵袭行为的抑制作用。结论 EMAP II抑制U251细胞的迁移和侵袭,其机制可能与Fox O1的表达上调有关。     

低强度脉冲电磁场对大鼠骨质疏松的影响

目的：观察低强度脉冲电磁场（pulsed electromagnetic fields，PEMFs）对去卵巢诱导骨质疏松症的大鼠生化指标和骨应力的影响。方法：雌性SD大鼠30只，随机等分为3组（n=10），分别为假手术组（Sham）、骨质疏松模型组（Model）、脉冲电磁场照射组（PEMFs）。经适应4wk后，在25mg·Kg^-1戊巴比妥钠腹腔麻醉下，Model组和PEMFs组摘除双侧卵巢．Sham组找到但不切除卵巢。各实验组均在相同环境下饲养，模型制备4wk后开始治疗，由GZY型低强度低频率脉冲电磁场发生仪产生低频脉冲磁场，根据实验要求，我们使用亥姆霍兹线圈形成均匀磁场，PEMFs组经照射刺激治疗，频率14．3Hz，场强2Gs．日照8h。Model组和Sham组正常饲养。治疗8wk后，对各组大鼠血清、尿液中ALP和Ca以及骨应力进行检测。结果：（1）ALP、Ca检测结果：与Model组相比，PEMFs组ALP值、Ca值差异均有统计学意义（P〈0．05）。其中，血清中ALP值，Model组为（275．16±228，57），PEMFs组为（179．30±87．68）；Ca值，Model组为（2，66±0，13），PEMFs组为（2．52±0．05）。（2）骨应力检测结果：Model组为（923．60±34．15Pa），PEMFs组为（1152．85±118，20Pa），组间差异有统计学意义（P〈0．05）。结论：研究发现，PEMFs对于促进骨重建、提高Ca吸收和骨应力恢复具有积极作用。     

ELF脉冲电磁场对脑组织钙离子迁移量的影响

目的研究极低频(extremely low frequency,ELF)脉冲电磁场对脑组织钙离子迁移量的影响。方法以鸡脑组织为生物学实验对象,以同位素示踪法为标记方法,用液体闪烁计数器为检测仪器,实验观察ELF脉冲电磁场对脑组织钙离子迁移量的影响。结果不同特征外加ELF脉冲电磁场对脑组织钙离子迁移量影响不同,如频率为16 Hz、场强为42.5 V/m的ELF脉冲电磁场可引起脑组织钙离子迁移量显著变化。频率为16 Hz、场强为62.2 V/m与频率为31 Hz、场强为42.5 V/m的ELF脉冲电磁场却未引起脑组织钙离子迁移量显著改变。结论只有频率和强度恰当组合的ELF电磁场才可引起脑组织钙离子迁移量显著升高,呈现出生物学窗效应。另一方面在既注重物理学分析又注重生物系统特性基础上,从膜通道内的带电粒子动力学方程入手分析讨论生物学窗效应产生机制。     

尘螨抗原对支气管上皮细胞单核细胞趋化蛋白-1表达的影响

目的探讨屋尘螨（Dermatophagoides pteronyssinus，Derp）抗原对支气管上皮细胞单核细胞趋化蛋白-1（MCP-1）表达的影响。方法使支气管上皮细胞BEAS-2B暴露于系列不同浓度（0．02、0．2．2、20μg／ml）的Derp抗原24h至96h，分别观察各时间点细胞的表现，然后用酶联免疫法（ELISA）检测其细胞上清MCP-1的浓度表达。结果正常为未加Derp抗原。表现为单层细胞完全平铺；实验各组在抗原的刺激下，表现为随着浓度和时间的增加，细胞逐渐变瘦长，细胞间距逐渐增大；在无抗原刺激因素培养条件下的对照组，MCP-1释放水平很低，而在加入Derp抗原组，引起细胞分泌MCP-1蛋白水平的显著增加，并随着时间和浓度增加，MCP-1蛋白的表达水平呈上升趋势，特别是在高浓度组，即20μg／ml抗原组，各时间点MCP-1的表达差异均有统计学意义（P〈0．01）。结论Derp抗原刺激气道上皮细胞，引起支气管上皮损伤和脱落等气道炎症反应，激发单核巨噬细胞产生大量的MCP-1，促成和加重气道炎症反应，因此认为MCP-1可能参与哮喘疾病的某些过程。     

Serum MCP-1 levels are increased in mild cognitive impairment and mild Alzheimer's disease

Upregulation of a number of chemokines, including monocyte chemotactic protein-1 (MCP-1), is associated with Alzheimer's disease (AD) pathological changes. Emerging evidence suggests that inflammatory events precede the clinical development of AD, as cytokine disregulation has been observed also in patients with mild cognitive impairment (MCI).

MCP-1 levels were evaluated in serum samples from 48 subjects with MCI, 94 AD patients and 24 age-matched controls. Significantly increased MCP-1 levels were found in MCI and mild AD, but not in severe AD patients as compared with controls. mRNA levels in peripheral blood mononuclear cells (PBMC), evaluated by quantitative RT-PCR analysis, paralleled serum MCP-1 levels. Moreover, a progressive MCP-1 decrease was observed over a 1-year follow up in a subgroup of MCI subjects converted to AD. MCP-1 upregulation is likely to be a very early event in AD pathogenesis, by far preceding the clinical onset of the disease. Nevertheless, as MCP-1 is likely to play a role in several pathologies with an inflammatory component, a possible usfulness as an early AD biomarker would be possible only in combination with other molecules.     

Involvement of MCP-1 in Tubulointerstitial Fibrosis Through Massive Proteinuria in Anti-GBM Nephritis Induced in WKY Rats

We investigated participation of monocyte chemoattractant protein-1 (MCP-1) in tubulointerstitial fibrosis and correlation between MCP-1 and proteinuria in Wistar-Kyoto (WKY) rats with glomerulonephritis induced by anti-glomerular basement membrane (anti-GBM) antibody. WKY rats showed marked proteinuria and severe glomerular crescent formation at 7 days post antibody injection. At 28 days, tubulointerstitial fibrotic lesions were observed, followed by sustained heavy proteinuria and severe tubulointerstitial fibrosis at 56 days. Histological examination revealed that the overlapped immunoreactivities of MCP-1, rat albumin, and p65NF-κB were detected in the same tubular segments of nephritic kidney, and a significant positive correlation was observed between proteinuria and MCP-1 expression in the tubulointerstitial fibrosis. ED-1- and CD8-positive cells were also abundant, and there was a good correlation between monocyte/macrophage recruitment and MCP-1 expression in the tubulointerstitial area. These results suggest that MCP-1 participates in the progression of tubulointerstitial fibrosis, through massive albuminuria, which is accompanied by marked monocyte/macrophage recruitment.     

腹膜间皮细胞在小鼠子宫内膜细胞刺激腹腔微环境中作用

目的：探讨腹腔微环境受子宫内膜细胞刺激改变中腹膜间皮细胞的作用及其对子宫内膜异位症发病机制的意义。方法： 注射小鼠子宫内膜上皮和间质细胞入小鼠腹腔，酶联免疫吸附法（ELISA）检测注射后4、24和72 h时点腹腔冲洗液细胞因子MCP-1/JE、IL-1α和IL-6蛋白表达，同步逆转录-聚合酶链式反应技术 (Real-Time RT-PCR) 检测腹膜组织 (主要含腹膜间皮细胞) 和腹腔巨噬细胞的细胞因子MCP-1/JE、IL-1 α和IL-6 mRNA表达。结果: 子宫内膜细胞刺激腹腔细胞因子蛋白含量迅速一过性升高，4 h时点为表达高峰，腹膜细胞因子基因表达与腹腔液蛋白表达同步，腹腔巨噬细胞基因表达高峰滞后于腹腔液蛋白表达。子宫内膜上皮细胞刺激腹腔炎症反应作用强于间质细胞。结论: 子宫内膜细胞刺激腹腔发生非特异性炎症反应，腹膜间皮细胞可能是其细胞因子效应的主要来源，提示腹膜在子宫内膜异位症中除作为病灶依附体外，还可能在腹腔微环境无菌性炎症反应中起重要作用。     

不同频率脉冲电磁场对人卵巢癌SKOV3细胞增殖、凋亡及迁移的影响

研究不同频率的脉冲电磁场(PEMF)对人卵巢癌SKOV3细胞增殖、凋亡和迁移的影响,为PEMF治疗或预防卵巢癌切除术后发生骨质疏松患者的安全性提供实验参考数据。分别对体外培养的人SKOV3细胞施加不同频率(8、16、32和64Hz)、固定强度为1mT的PEMF,2次/d,30min/次,间隔12h,共3d,以未施加PEMF的细胞作为对照组。分别采用EdU荧光法、Annexin V-FITC荧光法和划痕实验检测SKOV3细胞增殖、凋亡及迁移的情况。结果:频率8Hz的PEMF能明显抑制SKOV3细胞的增殖,并诱导其凋亡。频率8Hz和32Hz的PEMF能明显促进人卵巢癌细胞SKOV3的迁移,而频率16Hz的PEMF却明显地抑制SKOV3细胞的迁移。提示不同频率的PEMF对人卵巢癌细胞SKOV3增殖、凋亡及迁移的影响不同。在临床中对于卵巢癌切除术后的骨质疏松患者,应当慎重考虑PEMF治疗的安全性,并严格选择治疗参数。     

Decibel Attenuation of Pulsed Electromagnetic Field (PEMF) in Blood and Cortical Bone Determined Experimentally and from the Theory of Ohmic Losses

We studied the PEMF power attenuation in tissues representative of clinical applications (blood and cortical bone) to determine the amount of power available for PEMF purported biological effects. The experimental system consisted of a pair of nearly circular, parallel and coaxial coils separated by a distance of one coil diameter. The power attenuation was measured using a small search coil connected to a digital oscilloscope. The coils were powered by a voltage switch operating at two different frequencies (3.8 and 63 kHz) producing bursts of pulses (numbering 21 and 1619) and triggered at two different frequencies (1.5 and 15 Hz, respectively). The tissue samples were placed inside the coils so as to expose them to either transverse electric field (at the center of coils) or the transverse magnetic field (at the coil wire). The cylindrical coil geometry yielded closed-form expressions for power attenuation based on magnetic diffusion equation and ohmic losses due to bulk tissue magnetic permeability and electrical conductivity. The measured power attenuation at these PEMF frequencies of not more than one decibel (1 dB) was well explained by the theory for the 3.8 kHz but less so for the 63 kHz frequency PEMF. The results provide important insights regarding physical mechanism of weak PEMF power dissipation in tissues.     

MCP-1在胚胎种植中的作用

趋化因子是一类对中性粒细胞、单核细胞、嗜碱性粒细胞等起趋化和激活作用的多功能糖蛋白,介导炎症和免疫反应,单核细胞趋化蛋白-1（MCP-1）属于CC类趋化因子家族的一员。近年来研究表明,在激素的调节作用下,MCP-1在胚胎种植中发挥重要作用,其机制主要为趋化并激活单核巨噬细胞,参与Th2型免疫偏移及促进血管生成。     

白藜芦醇对TNF-α诱导的离体大鼠肺动脉内皮细胞损伤及MCP-1表达的影响

目的:探讨中药单体白藜芦醇(resveratrol,Res)对肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)诱导的离体大鼠肺动脉内皮细胞(rat pulmonary artery endothelial cells,RPAECs)中单核细胞趋化蛋白1(monocyte chemoattractant protein-1,MCP-1)的作用。方法:采用组织贴块法培养RPAECs,随机分为空白对照组(control组)、溶剂对照组(solvent组)、TNF-α(10μmol/L)组和Res(50μmol/L)+TNF-α组(Res组),每组又分成1、4和8 h三个时点(n=6),在8 h增加TNF-α+C1142(MCP-1特异性中和抗体)组(C1142组)。采用Western blot方法检测RPAECs中MCP-1蛋白表达,real-time PCR方法检测MCP-1 mRNA表达。结果:C1142可显著减少TNF-α诱导的RPAECs凋亡(P0.05)。相同时点solvent组的MCP-1蛋白和control组比较差异无统计学显著性;TNF-α组的MCP-1蛋白及mRNA较control组增高(P0.05);Res组的MCP-1蛋白及mRNA表达较TNF-α组降低(P0.05)。结论:MCP-1参与TNF-α诱导的RPAECs损伤;Res可降低TNF-α诱导的RPAECs中MCP-1表达,从而减少内皮细胞损伤。     

糖化白蛋白对人脐静脉内皮细胞MCP-1表达的影响及其机制研究

目的:探讨糖化白蛋白对内皮细胞中单核细胞趋化蛋白-1(MCP-1)表达的影响及其机制。方法:将人脐静脉内皮细胞(HUVECs)与不同浓度的糖化白蛋白共同培养,并用糖基化产物抑制剂氨基胍(AG)和抗氧化剂N-乙酰半胱氨酸(NAC)干预。分别用免疫细胞化学和夹心ELISA方法测定细胞MCP-1的表达,硫代巴比妥酸法和黄嘌呤氧化酶法测定细胞内丙二醛含量和超氧化物歧化酶活性。结果:糖化白蛋白促进HUVECs合成和分泌MCP-1。免疫细胞化学显示,HUVECs暴露于50mg/L糖化白蛋白后,随作用时间的延长(4 h、8 h、12h),MCP-1的表达增高(P0.01),分别为对照组的1.3、1.9和2.8倍(P0.01);糖化白蛋白能引起细胞内超氧化物歧化酶活性下降(P0.05)和丙二醛含量升高(P0.01)。氨基胍和N-乙酰半胱氨酸能抑制糖化白蛋白刺激内皮细胞表达MCP-1(P0.01),N-乙酰半胱氨酸能抑制糖化白蛋白对内皮细胞内超氧化物歧化酶和丙二醛的影响(P0.05)。结论:糖化白蛋白可刺激人类内皮细胞表达MCP-1,糖化白蛋白刺激MCP-1表达与其诱导细胞内氧化应激有关。     

No association of the MCP-1 promoter A-2518G polymorphism with bipolar disorder in the Korean population

It has been suggested that bipolar disorder is associated with altered immune function. Monocyte chemoattractant protien-1 (MCP-1) is a chemokine that influences both neural and immune functions. We thus hypothesized that MCP-1 may be related to the development or pathophysiology of bipolar disorder. In this case–control study, we investigated the association between the A-2518G single nucleotide polymorphism (SNP) of the MCP-1 promoter and bipolar disorder. Patients with bipolar disorder (n = 183; bipolar I = 145, bipolar II = 38) and healthy controls (350) were recruited for the study. No significant allelic or genotypic association was detected between the A-2518G polymorphism and any sample of bipolar disorder patients. When we pooled the healthy controls and the cases of bipolar I disorder from previous Korean studies and this study, we again found no significant association. No significant difference in either allele frequency or genotype distribution was observed between bipolar I and bipolar II disorders. There was no difference in the age at onset of bipolar disorder among the three genotype groups. Our data suggest that the A-2518G polymorphism of MCP-1 is not a major susceptibility factor for bipolar disorder in the Korean population. However, the physiological role of MCP-1 is highly suggestive of its being associated with bipolar disorder, and further analyses of other SNPs of MCP-1 remain to be performed.