血管紧张素-(1-7)对血管紧张素Ⅱ诱导的脐静脉内皮细胞MCP-1和ICAM-1表达的影响

目的： 研究血管紧张素-(1-7)［Ang-(1-7)］对血管紧张素Ⅱ（AngⅡ）诱导的脐静脉内皮细胞（HUVEC）单核细胞趋化蛋白-1（MCP-1）和细胞间黏附分子-1（ICAM-1）的影响,阐明Ang-(1-7)对AngⅡ在炎症方面的拮抗作用。方法: 体外培养HUVEC,随机分为：对照组;AngⅡ组;Ang-(1-7)组;AngⅡ+Ang-(1-7)组;AngⅡ+ Ang-(1-7) + Ang-(1-7)受体阻断剂A-779组。以ELISA法和半定量RT-PCR法从蛋白和mRNA水平检测MCP-1和ICAM-1的表达情况。结果: 与对照组比,AngⅡ（100 nmol/L）使MCP-1和ICAM-1的蛋白和mRNA表达明显增加（P<0.05）;Ang-(1-7)（1 000 nmol/L）使MCP-1和ICAM-1的蛋白和mRNA表达降低（P<0.05）;混合刺激组中,与AngⅡ组比较,Ang-(1-7) （10 nmol/L、100 nmol/L、1 000 nmol/L、10 000 nmol/L）呈剂量依赖性地抑制AngⅡ诱导的HUVEC MCP-1、ICAM-1蛋白和mRNA的表达（P<0.05）,Ang-(1-7) 浓度为1 000 nmol/L时,虽然蛋白和mRNA表达仍高于对照组,但无显著差异（P>0.05）;加入 A-779 组与AngⅡ组比较无显著差异（P>0.05）。结论: Ang-(1-7) 通过其特异性受体MAS拮抗AngⅡ诱导的HUVEC MCP-1和ICAM-1的表达,并呈浓度依赖性。     

青藤碱抑制TNF-α诱导人脐静脉内皮细胞VCAM-1表达

目的： 研究青藤碱（SN）对肿瘤坏死因子-α（TNF-α）诱导脐静脉内皮细胞（HUVECs ）VCAM-1表达的影响。方法： 从新鲜脐带中分离培养HUVECs。用TNF-α诱导HUVECs表达VCAM-1，实验组加入不同浓度的SN(0.25、0.5和1.0 mol/L)或地塞米松（1.0×10^-6 mol/L）进行干预，培养12 h后收获细胞，用实时定量PCR检测VCAM-1 mRNA的表达，用流式细胞仪检测细胞表面VCAM-1表达。结果： TNF-α可诱导VCAM-1 mRNA和VCAM-1的表达。进行药物干预后，各干预组相对VCAM-1 mRNA表达有不同程度下降(P<0.05)。SN（1.0 mol/L）和SN（0.5 mol/L）干预组细胞表面VCAM-1表达下降(P<0.05)。SN（0.25 mol/L）和地塞米松（1.0×10^-6 mol/L）干预组未显示对TNF-α 诱导的VCAM-1表达有抑制作用。结论： SN可抑制TNF-α诱导的脐静脉内皮细胞VCAM-1的表达。     

血管紧张素Ⅱ对单核细胞趋化因子及粘附分子表达的影响

为了观察血管紧张素Ⅱ（AⅡ）对人单核细胞株THP-1分泌单核细胞趋化因子（MCP-1）、人类脐静脉内皮细胞（HUVEC）分泌细胞间粘附分子（ICAM-1）的影响及AⅡ对HUVEC与THP-1的粘附功能的影响。我们利用ELISA法检测AⅡ作用后THP-1分泌MCP-1及HUVEC分泌ICAM-1的表达，利用计数法观察经AⅡ作用的HUVEC与THP-1的粘附率。结果发现四种不同浓度的AⅡ（10^-6,10^-7,10^-8,10^-9mol/L)刺激THP-1及HUVEC24h后，MCP-1及ICAM-1的分泌明显增加，AⅡ（10^-6mol/L)作用HUVEC6h后，其对THP-1细胞粘附率明显增加。表明AⅡ具有调节THP-1分泌MCP-1及HUVEC分泌ICAM-1的作用。AⅡ可通过致炎症作用参与动脉粥样硬化的发病过程。     

血管生长素-1基因修饰骨髓间充质干细胞对黏附因子表达的影响

目的探讨血管生长素1(Ang1)基因转导的骨髓间充质干细胞(Ang1-rMSC)对内皮细胞黏附因子表达的影响。方法经慢病毒载体介导构建Ang1-rMSC,观察VEGF刺激下不同时间点人脐静脉血管内皮细胞(HUVEC)黏附分子VCAM-1和ICAM-1表达的动态改变。采用RT-PCR及Western blot技术检测黏附分子mRNA及蛋白质水平的表达变化。结果HUVEC在20μg/L VEGF刺激下,ICAM-1和VCAM-1 mRNA水平显著升高,8h达到峰值,与对照组相比分别增加4.2倍及3.2倍。不同浓度上清液孵育的HUVEC在VEGF刺激后,ICAM-1和VCAM-1 mRNA表达均较对照组下降;含70%Ang1-rMSC上清液孵育的HUVEC,VCAM-1与ICAM-1表达较单纯VEGF组显著下降。结论Ang1-rMSC上清液可抑制VEGF诱导的HUVEC黏附分子表达,通过Ang1基因修饰Ang1-rMSC用于抑制干细胞移植伴发的炎症反应具有可行性。     

AngⅡ诱导血管内皮细胞衰老及凋亡相关基因的表达

目的： 探讨血管紧张素Ⅱ（AngⅡ）诱导人脐静脉内皮细胞(HUVECs)衰老及凋亡相关基因Bcl-2、Bax的表达。方法: 体外培养人脐静脉内皮细胞, 采用四甲基偶氮唑蓝比色法测定内皮细胞存活率，用AngⅡ(10^-6mol/L)及valsartan（AngⅡ1型受体特异性拮抗剂）干预,分为实验对照组、AngⅡ诱导组及valsartan组，采用β-半乳糖苷酶染色和流式细胞术鉴定细胞衰老，通过Hoechst33258荧光染色观察细胞形态学变化，并利用免疫细胞化学染色法、RT-PCR法和Western blotting分析各组细胞凋亡相关基因Bcl-2、Bax mRNA及蛋白的表达水平。结果: 与对照组相比，10^-6mol/L AngⅡ诱导组存活的细胞数为对照组的（81.90±0.04)%; (80.10±6.81)%的细胞呈现β-半乳糖苷酶阳性染色。流式细胞仪检测细胞周期停滞于G₀-G₁［(91.36±6.45)%］，证实细胞衰老；荧光显微镜可见明显的细胞凋亡［(31.84±2.86)%］。与AngⅡ诱导组相比，valsartan组Bcl-2mRNA及蛋白表达水平明显增高(P<0.05), Bax mRNA及蛋白表达水平降低（P<0.05）。结论: AngⅡ可诱导体外培养的HUVECs老化。经AngⅡ诱导的衰老HUVECs发生凋亡，提示细胞凋亡参与了AngⅡ诱导HUVECs细胞的衰老过程。AngⅡ诱导血管内皮细胞衰老的分子机制之一可能与Bcl-2、Bax mRNA及蛋白表达的失衡有关。缬沙坦对血管内皮细胞衰老有一定保护作用。     

NF-κB信号通路介导AngⅡ诱导THP-1巨噬细胞表达MMP-9

目的： 探讨血管紧张素Ⅱ（AngⅡ）诱导人单核/巨噬细胞（THP-1细胞）基质金属蛋白酶-9（MMP-9）表达的机制。方法: 用0.1 μmol/L佛波酯（PMA）作用48 h,诱导THP-1单核细胞分化为巨噬细胞后,随机分为4组：（1）PMA组,即对照组;（2）PMA+AngⅡ组（10-7mol/L,1 h）;（3）PMA+AngⅡ+2-硫代氨基甲酸吡咯烷组［PDTC（10 μmol/L,30 min）］;（4）PDTC组。用Western blotting蛋白印记技术分别检测总蛋白MMP-9及磷酸化核转录因子-κB p65（NF-κB p65）的变化,用RT-PCR检测MMP-9 mRNA表达的变化。结果: 与对照组相比,AngⅡ诱导THP-1巨噬细胞磷酸化NF-κB p65增加（1.02±0.10,P<0.05）,MMP-9蛋白量也增加（1.06 ±0.11,P<0.05）,MMP-9 mRNA表达上调（1.22±0.08,P<0.05）,而使用NF-κB的抑制剂PDTC后磷酸化NF-κB p65（0.99±0.12,P<0.01）减少,MMP-9表达明显受到抑制（1.04±0.14, P<0.01）,MMP-9 mRNA表达下调（0.90±0.06, P<0.01）。结论: NF-κB信号转导途径是AngⅡ诱导THP-1巨噬细胞表达MMP-9的重要途径之一。     

人参皂苷Rb1减轻人脐静脉内皮细胞氧化损伤

目的探讨人参皂苷Rb1(gRb1)对过氧化氢(H_2O_2)致人脐静脉内皮细胞(HUVECs)氧化损伤的影响。方法将HUVECs分为对照组、H_2O_2(20、40、80和160μmol/L)组、gRb1(10、20和40μmol/L)干预组。MTT法测定细胞存活率;annexin V-FITC/PI双染法检测凋亡;黄嘌呤氧化酶法测定SOD1活性;硫代巴比妥比色法计算MDA含量;Western blot测定细胞间黏附分子-1(ICAM-1)和血管细胞黏附分子-1(VCAM-1)蛋白表达。结果与对照组相比,H_2O_2呈浓度依赖性抑制细胞存活率(P0.05),增加细胞凋亡(P0.05),抑制SOD1活性(P0.05),增加MDA含量(P0.05),促进ICAM-1和VCAM-1蛋白表达(P0.05)。gRb1干预能够显著缓解上述指标的变化。结论 gRb1通过抑制细胞凋亡、改善氧化应激水平和抑制ICAM-1及VCAM-1蛋白表达减轻HUVECs氧化损伤。     

钙蛋白酶调节致心肌细胞肥大的信号转导

目的： 探讨钙敏感的calpain对血管紧张素Ⅱ（AngⅡ）介导的致心肌细胞肥大的钙调神经磷酸酶（CaN）信号通路的调节。方法: 原代培养的乳鼠心肌细胞为模型，AngⅡ刺激细胞Ca2+内流和（或）内贮细胞Ca2+释放， ［3H］-Leu掺入法反映心肌细胞蛋白合成速率，Fura-2/AM比率荧光成像分析细胞内钙信号变化，免疫印迹检测心肌细胞μ-calpain、m-calpain、CaN磷酸化及蛋白表达。结果: Ang Ⅱ呈浓度依赖性（10^-8-10^-5 mol·L^-1）促心肌细胞［Ca2+］i增加,各刺激组［Ca2+］i与对照组差异显著(P<0.01或P<0.05)。AngⅡ（10-7 mol·L^-1）剌激心肌细胞15 min后, μ-calpain的磷酸化明显增加，但其蛋白表达量无显著改变（P>0.05）, m-calpain蛋白表达及磷酸化量显著差异，CaN的蛋白表达量无显著改变，但其磷酸化增强, 环孢素A（CsA）抑制CaN的磷酸化。AngⅡ可使无血清培养新生大鼠心肌细胞［3H］-Leu掺入率随时间增加而增加，与对照组差异显著(P<0.01或P<0.05)，且在10^-8-10^-5 mol·L^-1范围内呈量效依赖关系。结论: AngⅡ剌激细胞外Ca2+跨膜内流和（或）内贮Ca2+的释放导致μ-calpain的激活，进一步激活钙敏感的CaN信号通路，在心肌肥厚的病理过程中起重要作用。     

梅毒螺旋体膜蛋白 Tpp17体外活化血管内皮细胞的实验研究北大核心CSCD

目的：研究梅毒螺旋体重组蛋白Tpp17体外对人脐静脉内皮细胞（ HUVEC ）的活化作用,探讨膜蛋白Tpp17在梅毒免疫学发病机制中的作用。方法将采用基因工程技术重组合成的梅毒螺旋体膜蛋白Tpp17刺激HUVEC, ELISA测培养上清中细胞因子TNF-α、MCP-1、ICAM-1及E-selectin的水平,荧光定量聚合酶链反应测HUVEC中TNF-α、MCP-1、ICAM-1及E-selectin mRNA的转录水平;将重组蛋白Tpp17预处理的HUVEC与Calcein-AM标记的THP-1细胞共培养,荧光倒置显微镜下观察HUVEC与THP-1细胞的黏附情况;将HUVEC接种于transwell小室的下室,重组蛋白Tpp17处理后,Calcein-AM标记的THP-1细胞接种于上室,荧光倒置显微镜观察下并计数THP-1细胞的迁移率。结果梅毒螺旋体膜重组蛋白Tpp17可上调HUVEC细胞因子TNF-α、MCP-1、ICAM-1及E-se-lectin的表达水平,提高其对THP-1细胞的趋化和黏附能力,与空白组比较,差异有统计学意义（ P＜0．05）。结论梅毒螺旋体膜重组蛋白Tpp17可体外活化HUVEC,提高HUVEC对THP-1细胞的趋化和黏附能力,可能在梅毒的免疫病理中起重要作用。     

载脂蛋白C3促进THP-1细胞向小鼠主动脉黏附

目的 探讨载脂蛋白C3(APOC3)对THP-1细胞与小鼠主动脉黏附效应的影响.方法 在无菌环境下应用外科显微技术分离C57BL/6小鼠主动脉,在体外给予APOC3刺激16 h后与标记有CFSE荧光的单核细胞来源THP-1细胞黏附1h,观察黏附效应的变化,同时应用免疫组化方法检测血管黏附分子1(VCAM-1)和细胞间黏附分子1(ICAM-1)的表达.结果 给予APOC3刺激的主动脉与对照主动脉相比,与THP-1细胞的黏附效应增加,VCAM-1和ICAM-1表达上调,且前者比后者上调显著.结论 载脂蛋白C3促进THP-1细胞向小鼠主动脉黏附.     

Role of Tyrosine Kinase Enzymes in TNF-α and IL-1 Induced Expression of ICAM-1 and VCAM-1 on Human Umbilical Vein Endothelial Cells

The aim of this study was to analyse the potential roles of protein kinase enzymes in tumour necrosis factor-α (TNF-α) and interleukin-1 (IL-1) induced expression of the adhesion molecules intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) on human umbilical vein endothelial cells (HUVEC). The authors observed a marked increase in ICAM-1 and VCAM-1 expression on HUVEC stimulated for 24 h by TNF-α (10 ng/ml) or IL-1 (20 ng/ml). Pre-treatment of HUVEC for 30 min with protein tyrosine kinase (PTK) inhibitors genistein and herbimycin A (10 μg/ml and 0.5 μg/ml, respectively) before stimulation with IL-1 did not affect the expression of these molecules. Similar results were observed with respect to VCAM-1 expression on HUVEC stimulated by TNF-α. In contrast, pre-incubation of HUVEC with PTK inhibitors prior to the addition of TNF-α significantly enhanced subsequent expression of ICAM-1, although spontaneous expression of ICAM-1 on unstimulated HUVEC was unaffected. Western blot analysis demonstrated a significant increase in phosphorylated tyrosine protein levels in HUVEC stimulated by TNF-α , and significantly lower levels of these proteins in TNF-α stimulated HUVEC pre-treated with PTK inhibitors. These results demonstrate that IL-1 induced ICAM-1 and VCAM-1 expression does not result from activation of PTK-dependent pathways. In the case of TNF-α induced responses, the selective co-stimulatory effect of this cytokine in combination with PTK inhibitors on ICAM-1 expression suggests a complicated intracellular pathway of TNF-α induced ICAM-1 expression, possibly involving down-modulation of increases in ICAM-1 by PTK enzymes.     

轻度修饰低密度脂蛋白对血管内皮细胞粘附功能影响及其机理初探

目的:观察轻度修饰LDL(MM-LDL)对培养人脐静脉内皮细胞(HUVEC)与人类单核细胞系U937粘附功能及其表面粘附分子表达的影响。方法:利用计数法观察经MM-LDL作用的HUVEC与U937细胞的粘附率;用ELISA方法检测MM-LDL作用后HUVEC膜表面粘附分子血管细胞粘附分子-1(VCAM-1)、细胞间粘附分子-1(ICAM-1)及P选择素(P-selectin)的表达。结果:MM-LDL(75mg/L)作用HUVEC4h后,其对U937细胞粘附率明显增加(P<0.01),HUVEC膜表面未见VCAM-1,ICAM-1,P-selectin表达上调,作为阳性对照重组肿瘤坏死因子α(rTNF_α)5.0μg/L可显著诱导以上3种粘附分子表达。延长MM-LDL与HUVEC作用时间至18h可诱导产生P-selectin表达,对VCAM-1表达无影响。结论:MM-LDL诱导的HUVEC与U937粘附不是通过ICAM-1、VCAM-1介导的,P-selectin可能起一定的作用。     

血管紧张素Ⅱ对THP-1源性泡沫细胞ATP结合盒转运子A1的影响

目的： 以THP-1源性泡沫细胞为研究对象，探讨血管紧张素Ⅱ（AngⅡ）对THP-1源性泡沫细胞ATP结合盒转运子A1(ABCA1)表达、细胞内胆固醇含量及胆固醇流出的影响。方法: 运用逆转录-多聚酶链反应（RT-PCR）和Western blotting分别检测AngⅡ对ABCA1 mRNA与ABCA1蛋白表达的影响，采用酶法，通过荧光分光光度计检测细胞内胆固醇含量，应用液体闪烁计数仪检测胆固醇流出的变化。结果: AngⅡ能引起THP-1源性泡沫细胞胆固醇含量显著升高(P<0.05)、ABCA1表达显著减少（P<0.05），AngⅡ受体拮抗剂厄贝沙坦（Irb）能显著减少细胞内胆固醇含量(P<0.05)、促进细胞内胆固醇流出及减轻AngⅡ对ABCA1的抑制作用(P<0.05)。结论: AngⅡ有通过其受体抑制ABCA1表达，促进泡沫细胞形成，加速动脉粥样硬化的作用。     

通心络超微粉对高脂饮食兔主动脉内皮保护机制的实验研究

目的： 探讨通心络超微粉对高脂饮食兔主动脉内皮损伤的干预作用及其可能机制。方法： 健康雄性新西兰白兔32只随机分为空白对照组、模型组、阿托伐他汀组、通心络组4组。空白对照组饲以普通饲料；模型组饲以高脂饲料；阿托伐他汀组饲以高脂饲料同时阿托伐他汀3 mg·kg^-1·d^-1灌胃；通心络组饲以高脂饲料同时通心络超微粉0.31 g·kg^-1·d^-1灌胃，连续给药，于6周末酶法检测各组血脂水平，比色法检测各组血清NO、MDA水平及SOD活性，免疫组织化学染色法检测主动脉内皮细胞NF-κB核转位情况及ICAM-1 蛋白表达，RT-PCR法检测ICAM-1 mRNA表达。结果：模型组血清TC、TG、LDL-C、HDL-C及MDA水平均显著高于空白对照组(P<0.01)，NO水平及SOD活性低于空白组(P<0.01)；主动脉内皮细胞NF-κB核转位、ICAM-1基因及蛋白表达明显多于空白组（P<0.01）。两药物干预组TC、TG、LDL-C及MDA水平均显著低于模型组(P<0.01)，NO水平和SOD活性则高于模型组（P<0.05, P<0.01），主动脉内皮细胞NF-κB核转位、ICAM-1基因及蛋白表达亦明显少于模型组（P<0.05，P<0.01），通心络组HDL-C显著高于模型组（P<0.05），且通心络组对上述指标的作用均优于阿托伐他汀组（P<0.05，P<0.01）。结论： ① 高脂血症可使血管内皮细胞受损，其机制可能与氧化应激、NF-κB的激活及ICAM-1基因与蛋白的表达异常有关。② 通心络超微粉可在一定程度上减轻上述病理损伤，可能与其抗氧化、抑制NF-κB核转位进而降低ICAM-1基因及蛋白表达有关。     

Expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in human crescentic glomerulonephritis

AIMS: In glomerulonephritis, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) may play important roles in the formation of crescents. These studies are designed to evaluate the expression patterns of ICAM-1 and VCAM-1 in human crescentic glomerulonephritis and to determine the cellular origin of adhesion molecules in the crescentic lesions. METHODS AND RESULTS: We examined the expression of ICAM-1 and VCAM-1 proteins in renal biopsies with cellular (n=7), fibrocellular (n=9) or fibrous (n=4) crescentic glomerulonephritis, and six controls by immunohistochemistry. mRNA expression of ICAM-1 and VCAM-1 was further evaluated by RNA in-situ hybridization. Cytokeratin or CD68 immunohistochemistry was performed on the same sections, where in-situ hybridization had been carried out. In cellular crescents, ICAM-1 and VCAM-1 proteins were over-expressed to a similar extent. Of the three types of crescents, the extent of ICAM-1 immunopositivity was the greatest in the cellular crescents and decreased towards the fibrous crescents (P < 0.05). Yet the extent of VCAM-1 immunoreactivity was not different between the types. Fibrous crescents still contained some epithelial cells and showed only VCAM-1 expression. In the glomeruli with cellular or fibrocellular crescents, the extent of ICAM-1 immunopositivity in the glomerular tufts was significantly larger than that of VCAM-1 (P < 0.05). In an in-situ hybridization study, the mRNA expression patterns of ICAM-1 and VCAM-1 paralleled their protein expressions. A double-labelling study showed that the signal for ICAM-1 and VCAM-1 mRNAs was mainly present in cytokeratin-positive and CD68-negative cells in the crescentic lesions. CONCLUSIONS: These results suggest that glomerular parietal epithelial cells in cellular crescents up-regulate both ICAM-1 and VCAM-1, and that some epithelial cells retained in fibrous crescents persistently over-express VCAM-1, but not ICAM-1. They also suggest that ICAM-1 is involved in early leucocyte recruitment into glomeruli in crescentic glomerulonephritis.     

Differential influences of bFGF and VEGF on the expression of vascular cell adhesion molecule-1 on human umbilical vein endothelial cells]

A fundamental feature of inflammation includes angiogenesis, adhesion of leukocytes to vascular endothelium, and entry of leukocytes into inflamed tissues. Recent studies have suggested that angiogenesis and cellular adhesion may be mutually linked processes. Both basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) have been shown to facilitate angiogenesis. However, their roles in the expression of adhesion molecules on the endothelial cells have not been clarified. The current studies therefore examined the effect of bFGF and VEGF on the expression of vascular cell adhesion molecule-1 (VCAM-1) on human umbilical vein endothelial cells (HUVEC) stimulated with tumor necrosis factor alpha (TNF-alpha). HUVEC (1 x 10(4)/well) were incubated in a 96 well microtiter plate with culture medium containing endothelial cell growth supplement (ECGS) for 24 h. After the incubation, culture medium was replaced by ECGS free culture medium with or without TNF-alpha (10 ng/ml), bFGF (10 ng/ml) and VEGF (10 ng/ml), and the culture was further carried out for additional 24 h. The expression of VCAM-1, E-selectin, and intercellular adhesion molecule-1 (ICAM-1) was measured by cell ELISA and the proliferation of HUVEC was measured by MTT colorimetric assay. Soluble VCAM-1 (sVCAM-1) in the supernatants were assessed by ELISA. Although, both bFGF and VEGF supported the proliferation of HUVEC, bFGF, but not VEGF, selectively suppressed the expression of VCAM-1 on HUVEC stimulated with TNF-alpha. The expression of ICAM-1 and E-selectin induced by TNF-alpha was not inhibited by either bFGF or VEGF. In addition, bFGF also decreased the levels of sVCAM-1 in the supernatants of TNF-alpha stimulated HUVEC. The data indicate that bFGF, but not VEGF, suppresses the production of VCAM-1 by HUVEC under stimulation with TNF-alpha. These results therefore suggest that angiogenic cytokines bFGF and VEGF play different roles in the regulation of the expression of adhesion molecules on endothelial cells under inflammation.     

尼古丁对中性粒细胞活化及细胞间粘附分子基因表达的影响

目的:观察尼古丁对中性粒细胞(PMNs)的活化,PMNs与内皮细胞的粘附及内皮细胞表达ICAM-1mRNA,有助于阐明尼古丁在慢性阻塞性肺疾患(COPD)炎症发病中的作用。方法:测定β-葡萄糖醛酸苷酶及溶菌酶活性,以反映PMNs的活化;培养人脐静脉内皮细胞,观察PMNs与内皮细胞的粘附;制备探针,提取总RNA,Northern杂交测细胞间粘附分子-1(ICAM-1)mRNA。结果:尼古丁可活化PMNs,增加PMNs－内皮细胞粘附;增强ICAM-1mRNA表达,764-3可明显抑制尼古丁的上述作用。结论:尼古丁通过活化PMNs,促进PMNs－内皮细胞粘附,在COPD慢性炎症发病中起重要作用。而这种粘附作用的增加与粘附分子表达增强有关;抑制尼古丁的上述作用可能是764-3抗炎作用的部分机理。     

Expression of vascular cell adhesion molecule-1 in fibroblastlike synoviocytes after stimulation with tumor necrosis factor.

Rapid expression of mRNA encoding vascular cell adhesion molecule-1 (VCAM-1) was induced by tumor necrosis factor (TNF) in fibroblast-like cells obtained from synovial tissue. Both alternatively spliced forms of VCAM-1 mRNA were detected by polymerase chain reaction in TNF-stimulated fibroblast-like synoviocytes. Western blotting analysis showed that two distinct proteins, reactive with an anti-VCAM-1 anti-sera, were expressed by 2 hours of TNF stimulation in both synoviocytes and human umbilical cord vein endothelial cells (HUVEC). The majority of HUVEC and synoviocytes displayed VCAM-1 surface expression after several hours of TNF stimulation. In contrast, dermal fibroblasts upregulated intercellular adhesion molecule-1 (ICAM-1) but not VCAM-1 expression in response to TNF. These results indicate that VCAM-1 and ICAM-1 expression can be differentially regulated and suggest tissue specific regulation of VCAM-1 expression. Furthermore, these findings may provide an explanation for the chronic retention and activation of long-lived lymphocytes and monocytes, which express VLA-4 (the receptor for VCAM-1), in the synovium in rheumatoid arthritis.