系统性红斑狼疮患者皮损及血清黏附分子及一氧化氮水平的研究

目的：探讨血管细胞黏附分子-1（VCAM-1）、细胞间黏附分子-1（ICAM-1）、E-选择素水平及一氧化氮（N0）在皮损表达和血清水平与系统性红斑狼疮（SEE）活动性的关系。方法：①应用ELISA试剂盒测定血清细胞黏附分子。血清NO的检测使用NO硝酸还原酶试剂盒;②采用SABC免疫组化技术检测细胞黏附分子和NO的表达。结果：①血清中可溶性VCAM-1（sVCAM—Ⅰ）、可溶性ICAM-1（sICAM—Ⅰ）、NO水平病例组均较对照组明显升高（P均〈0．01）;病例组活动期较非活动期升高（P〈0．01）。②SLE活动期皮损处血管内皮细胞VCAM-1、ICAM-1、E-选择素和NO的表达与对照组比较．均明显上调（P〈0．05）。③SLE患者sICAM-1水平与SLE疾病活动指数（SLEDAI）有相关性（r=0．590,P=0．026）,NO水平与抗ds—DNA抗体呈正相关（r=0．777,P=0．001）。④血管内皮细胞内各黏附分子、NO表达的强度依次为VCAM-1〉ICAM—1〉NO〉E-选择素。结论：VCAM-1、ICAM-1、E-选择素和NO可能在SLE的发生、发展中起重要的组织损伤作用。     

系统性红斑狼疮患者皮损及血清粘附分子的检测

目的：探讨血管内皮细胞间黏附分子-1（ICAM-1）、细胞黏附分子-1（VCAM-1）、sE-选择素水平在皮损表达及血清水平与系统性红斑狼疮（SLE）活动性的关系。方法：用ELISA试剂盒测定血清细胞黏附分子,应用SABC免疫组化技术检测细胞黏附分子的表达。结果：实验组血清可溶性ICAM-1（sICAM-1）和可溶性VCAM—1（sVCAM-1）的水平均较高于对照组,差异有统计学意义（均P〈0．01）;实验组活动期ICAM-1水平较非活动期升高（P〈0．01）。SLE患者活动期皮损血管内皮细胞ICAM-1、VCAM-1、E-选择素的表达高于对照组,差异有统计学意义（P〈0．05）。SLE血管内皮细胞各黏附分子表达的水平依次为VCAM-1〉ICAM-1〉E-选择素。结论：VCAM-1、ICAM-1、E-选择素等粘附分子可能在SLE的发生、发展中起重要的组织损伤作用。     

白念珠菌对人血管内皮细胞表面黏附分子表达和细胞因子分泌的影响

目的 观察白念珠菌对人血管内皮细胞表面细胞间黏附分子-1(ICAM-1)、血管细胞黏附分子-1(VCAM-1)表达及分泌白介素6(IL-6)、白介素8(IL-8)水平的影响.方法 分离健康产妇胎盘脐静脉内皮细胞进行原代培养,细胞长至单层时,与白念珠菌共培养不同时间,再用逆转录-聚合酶链反应法分析内皮细胞表面ICAM-1mRNA、VCAM-1mRNA的表达,用ELISA法检测IL-6、IL-8分泌水平.结果 白念珠菌刺激4h后血管内皮细胞表达ICAM-1 mRNA、VCAM-1 mRNA明显增加,至8h达到峰值.而IL-6,IL-8的分泌在4h后也明显增加(P<0.05),24h到达高峰(P<0.01).结论 白念珠菌可诱导人血管内皮细胞ICAM-1 mRNA、VCAM-1 mRNA表达及促进IL-6、IL-8分泌.     

银屑病患者血清E、L、P选择素和VCAM-1的表达

目的:研究银屑病患者血清中E、P、L选择素及血管细胞黏附分子-1(VCAM-1)的表达及其临床意义。方法:用双抗体夹心酶联免疫吸附法测定银屑病进行期、静止期患者和正常对照组健康人血清E、L、P选择素和VCAM-1的水平。结果:(1)进行期组患者血清E、L、P选择素和VCAM-1水平显著高于静止期组和正常对照组(P值均<0.05)。(2)静止期组患者血清E、L、P选择素和VCAM-1水平与正常对照组无显著性差异(P值均>0.05)。(3)患者血清E、L、P选择素和VCAM-1与PASI评分均无显著相关性。结论:E、L、P选择素和VCAM-1在银屑病活动期的炎症细胞聚集和黏附过程中起着重要作用。     

梅毒螺旋体膜蛋白Tpp47对血管内皮细胞黏附功能影响的实验研究

【摘要】 目的 探讨梅毒螺旋体膜蛋白Tpp47对血管内皮细胞的作用。 方法 利用基因工程技术重组合成的梅毒螺旋体膜蛋白Tpp47及脂多糖分别刺激人脐静脉内皮细胞（HUVEC）,ELISA检测上清中细胞间黏附分子1（ICAM-1）及E选择素的水平;荧光定量PCR检测HUVEC中ICAM-1及E选择素 mRNA的转录水平;MTT法检测HUVEC增殖水平;将重组蛋白Tpp47及脂多糖预处理的HUVEC与钙黄绿素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。 结果 梅毒螺旋体膜重组蛋白Tpp47刺激HUVEC后,其分泌的黏附分子ICAM-1（1.28 ± 0.03）及E选择素（0.51 ± 0.01）水平显著提高,与空白对照组（0.90 ± 0.01与0.13 ± 0.03）比较,差异均有统计学意义（t值分别为18.28和18.19,均P ＜ 0.05）;将重组蛋白Tpp47刺激24 h后的HUVEC与THP-1细胞共培养,HUVEC与THP-1细胞的黏附率为56.1% ± 1.9%,较空白对照组（16.3% ± 2.1%）明显提高,差异有统计学意义（χ2 = 12.65,P ＜ 0.05）。MTT试验结果显示,重组蛋白Tpp47刺激HUVEC后,其增殖率（19.5% ± 1.7%）高于空白对照组（10.0% ± 3.1%）,差异有统计学意义（χ2 = 3.92,P ＜ 0.05）;较脂多糖低（41.2% ± 3.7%）,差异有统计学意义（χ2 = 10.42,P ＜ 0.05）。 结论 梅毒螺旋体膜重组蛋白Tpp47可体外上调HUVEC与THP-1的黏附能力及促进HUVEC增殖,可能在梅毒的发病中起一定作用。     

复方昆明山海棠对TNF-α诱导的人血管内皮细胞ICAM-1表达影响的试验研究

目的通过复方昆明山海棠对TNF-α诱导的人血管内皮细胞ICAM-1表达影响的研究,探讨复方昆明山海棠的抗炎机制。方法采用体外细胞培养、免疫组化技术观察复方昆明山海棠对TNF-α诱导的内皮细胞黏附分子ICAM-1表达的影响。结果复方昆明山海棠（0.05～2）mg/ml预处理30min,可不同程度地抑制TNF-α诱导的内皮细胞黏附分子ICAM-1的表达,与空白对照组相比较有统计学意义（P〈0.05）;到2mg/ml时ICAM-1的表达减到最弱（P〈0.01）。结论复方昆明山海棠能明显抑制TNF-α诱导的人血管内皮细胞表面黏附分子ICAM-1表达,（0.05-2）mg/ml范围内其抑制作用的强弱与浓度有关。提示复方昆明山海棠有可能是通过抑制人血管内皮细胞表面黏附分子ICAM-1的表达从而减少白细胞与血管内皮的黏附来发挥其抗炎作用的。     

玫瑰糠疹表皮内血管细胞黏附分子1和细胞间黏附分子1的表达

目的:研究血管细胞黏附分子(1VCAM-l)和细胞间黏附分子(1ICAM-1)在玫瑰糠疹发病中的作用。方法:采用链酶卵白素-过氧化物酶法检测表皮内VCAM-l、ICAM-1的表达。结果:VCAM-1在玫瑰糠疹皮损表皮基底层、棘层表达高于皮损周围正常皮肤及正常人皮肤,在皮损周围正常皮肤棘层表达高于正常人皮肤。ICAM-1在皮损、皮损周围正常皮肤基底层的表达高于正常人皮肤,在皮损棘层的表达高于皮损周围正常皮肤及正常人皮肤。结论:玫瑰糠疹表皮VCAM-1、ICAM-1的高表达可能与玫瑰糠疹的发病机制有关。     

SLE患者外周血单一核细胞介导T淋巴细胞对血管内皮细胞黏附的实验研究

目的 探讨活动期SLE患者外周血单一核细胞(PBMC)是否介导了T淋巴细胞对血管内皮细胞的黏附.方法 体外培养人脐静脉内皮细胞(HUVEC),活动期SLE PBMC培养上清液预刺激48 h,通过RT-PCR检测PBMC培养上清液诱导HUVEC黏附分子的表达;划痕实验检测PBMC培养上清液诱导内皮细胞运动迁移能力的改变;HUVEC与T细胞共培养,研究PBMC培养上清液预刺激是否影响T细胞对HUVEC的黏附.结果 HUVEC经过活动期SLE PBMC培养上清刺激后,内皮细胞黏附分子ICAM-1mRNA,VCAM-1 mRNA和E-钙黏蛋白mRNA表达明显增加,而IL-17抗体能明显抑制黏附分子的表达.黏附分子表达增加介导了内皮细胞运动能力增加,介导T细胞对内皮细胞的黏附,而IL-17中和抗体能抑制T细胞对内皮细胞的黏附和抑制内皮细胞的运动迁移.结论 活动期SLE PBMC培养上清液可以通过诱导血管内皮细胞黏附分子表达,黏附T细胞,介导狼疮血管炎的发生.     

SLE患者外周血单一核细胞介导T淋巴细胞对血管内皮细胞黏附的实验研究

目的 探讨活动期SLE患者外周血单一核细胞(PBMC)是否介导了T淋巴细胞对血管内皮细胞的黏附.方法 体外培养人脐静脉内皮细胞(HUVEC),活动期SLE PBMC培养上清液预刺激48 h,通过RT-PCR检测PBMC培养上清液诱导HUVEC黏附分子的表达;划痕实验检测PBMC培养上清液诱导内皮细胞运动迁移能力的改变;HUVEC与T细胞共培养,研究PBMC培养上清液预刺激是否影响T细胞对HUVEC的黏附.结果 HUVEC经过活动期SLE PBMC培养上清刺激后,内皮细胞黏附分子ICAM-1mRNA,VCAM-1 mRNA和E-钙黏蛋白mRNA表达明显增加,而IL-17抗体能明显抑制黏附分子的表达.黏附分子表达增加介导了内皮细胞运动能力增加,介导T细胞对内皮细胞的黏附,而IL-17中和抗体能抑制T细胞对内皮细胞的黏附和抑制内皮细胞的运动迁移.结论 活动期SLE PBMC培养上清液可以通过诱导血管内皮细胞黏附分子表达,黏附T细胞,介导狼疮血管炎的发生.     

SLE患者外周血单一核细胞介导T淋巴细胞对血管内皮细胞黏附的实验研究

目的 探讨活动期SLE患者外周血单一核细胞(PBMC)是否介导了T淋巴细胞对血管内皮细胞的黏附.方法 体外培养人脐静脉内皮细胞(HUVEC),活动期SLE PBMC培养上清液预刺激48 h,通过RT-PCR检测PBMC培养上清液诱导HUVEC黏附分子的表达;划痕实验检测PBMC培养上清液诱导内皮细胞运动迁移能力的改变;HUVEC与T细胞共培养,研究PBMC培养上清液预刺激是否影响T细胞对HUVEC的黏附.结果 HUVEC经过活动期SLE PBMC培养上清刺激后,内皮细胞黏附分子ICAM-1mRNA,VCAM-1 mRNA和E-钙黏蛋白mRNA表达明显增加,而IL-17抗体能明显抑制黏附分子的表达.黏附分子表达增加介导了内皮细胞运动能力增加,介导T细胞对内皮细胞的黏附,而IL-17中和抗体能抑制T细胞对内皮细胞的黏附和抑制内皮细胞的运动迁移.结论 活动期SLE PBMC培养上清液可以通过诱导血管内皮细胞黏附分子表达,黏附T细胞,介导狼疮血管炎的发生.     

Tumour necrosis factor alpha is pro-inflammatory in normal human skin and modulates cutaneous adhesion molecule expression

Tumour necrosis factor a (TNF-α) is a potent immunoregulatory cytokine produced by many cutaneous cells, including kcratinocytes, mast cells and Langerhans cells. To explore its potential role in inflammatory skin disease, we have studied immunohistochemically the effects of intradermal recombinant human TNF-α (rHuTNF-α) on cutaneous inflammatory cells, adhesion molecules and Langerhans cells in normal human skin. Volunteers received rHuTNF-α 100U (group A), 5000 U (group B), or 100 U daily for 5 days (group C), and biopsies were taken at 6 h (groups A and B), or 6 h after the final injection (group C). An inflammatory cell infiltrate developed in all cases: following single injections of either 100 or 5000 U rHuTNF-α this was predominantly neutrophilic, whereas following multiple injections of 100 U few neutrophils were seen, although many lymphocytes (CD3⁺, CD4⁴) were present. In all groups there was an increase in cells of monocyte/macrophage lineage (CD36⁾⁺. TNF-α induced a dose- and time-dependent decrease in CDla⁺ epidermal Langerhans cell numbers and an increase in dermal CDla⁴ cells, suggesting migration of Langerhans cells away from the epidermis. TNF-α induced endothelial E-selectin, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in all groups, and adhesion molecule expression by interstitial dermal dendritic cells (ICAM-1 and VCAM-1) and keratinocytes (ICAM-1) was observed. These findings indicate that TNF-α is a potent modulator of cutaneous immune function in vivo, and this central role in the cutaneous immune response suggests that TNF-α may be an attractive target for therapeutic inhibition.     

The effect of herpesvirus infection on the expression of cell adhesion molecules on cultured human dermal microvascular endothelial cells

Cell-mediated immune response to herpes simplex virus (HSV) may be important in the pathogenesis of herpes keratitis, erythema multiforme or Behcet's disease. We examined whether herpesvirus infection regulates the expression of cell adhesion molecules on cultured human dermal microvascular endothelial cells (HDMEC) and the regulation of T-lymphocytes binding to HDMEC. The expression of intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), or E-selectin on HDMEC increased significantly after treatment with HSV-1, HSV-2, or measles virus on HDMEC. Anti-IL-1 alpha antibody or anti-TNF alpha antibody partially inhibited the expression of ICAM-1, VCAM-1, or E-selectin on HDMEC. The binding of T-lymphocytes to HDMEC increased significantly after the treatment of HSV-1 or measles virus on HDMEC. The binding of T-lymphocytes to HDMEC was significantly inhibited after 16 h of incubation following treatment with anti-ICAM-1 antibody, anti-IL-1 alpha antibody or anti-TNF alpha antibody to HDMEC. These study results suggest that HSV induces the increased expression of ICAM-1, or induction of VCAM-1 and E-selectin on HDMEC and that among these adhesion molecules, the expression of ICAM-1 on HDMEC mainly regulates the binding of T-lymphocytes to HDMEC. The data also suggest that IL-1 alpha or TNF alpha which was produced by HSV infected HDMEC may be related to these events.     

云南马齿苋提取物的抗炎机制研究

目的 探讨马齿苋提取物对肿瘤坏死因子?琢（TNF-?琢）、细胞间黏附分子1（ICAM-1）表达的影响,初步阐明马齿苋抗炎机制。方法　建立二甲苯致小鼠耳廓肿胀炎症模型,并分为4组。空白组：不采用任何干预措施,阴性对照组：建立模型后,不加干预,阳性组：模型建立后,使用韩国进口马齿苋外涂小鼠致炎处,实验组：建立模型后,使用云南马齿苋提取物涂于致炎处。组织病理及免疫组化观察TNF-?琢、ICAM-1的变化并比较各组的抗炎效果。结果　组织病理：空白组：真皮可见血管及少量淋巴细胞;阴性对照组：真皮组织疏松、水肿及大量淋巴细胞浸润;阳性组：轻度水肿,真皮血管周围可见部分炎症细胞浸润;实验组：HE染色：真皮轻度水肿,血管周围可见部分炎症细胞浸润。免疫组化：空白组：TNF-?琢、ICAM-1未见表达;阴性对照组：TNF-?琢强阳性表达于血管内皮细胞膜,ICAM-1强阳性表达于血管内皮细胞膜及淋巴细胞膜;阳性组：TNF-?琢阳性表达于血管内皮细胞膜,ICAM-1阳性表达于血管内皮细胞膜及淋巴细胞膜;实验组： TNF-?琢阳性表达于血管内皮细胞膜,ICAM-1阳性表达于血管内皮细胞膜及淋巴细胞膜。秩和检验：实验组与空白组比较TNF-?琢、ICAM-1,P值均 < 0.01,差异有统计学意义。结论　云南马齿苋抗炎机制可能与影响TNF-?琢、ICAM-1的表达有关。     

The effect of methotrexate on the expression of cell adhesion molecules and activation molecule CD69 in psoriasis

BACKGROUND: The mechanism of the action of methotrexate (MTX) in the treatment of psoriasis has not been completely elucidated. OBJECTIVE: To assess the effect of MTX on the expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E-selectin, activation molecule CD69 and T-cell phenotype in skin specimens from patients with psoriasis. METHODS: We performed an immunohistochemical analysis of the expression of T-cell phenotype and cell adhesion/activation molecules in skin biopsies from patients with psoriasis treated with a fixed dose of MTX (12.5 mg/week). To determine data on the epidermal/dermal T-cell infiltration we carried out a manual quantification. RESULTS: Skin samples prior to therapy showed a moderate to severe inflammatory infiltrate, mainly due to T lymphocytes with a helper/inducer (CD4) phenotype. Most of these cells also expressed ICAM-1 and VCAM-1. Blood vessels showed expression of E-selectin and VCAM-1, and keratinocytes were positive for ICAM-1 staining. The cell infiltrate was reduced after therapy, as well as the expression of cell adhesion molecules. However, we also noted the persistence of the T lymphocyte phenotype CD8(+), expressing the CD69 activation molecule, after the MTX treatment. CONCLUSIONS: MTX downregulates the expression of some adhesion molecules, a phenomenon that may contribute to its anti-inflammatory therapeutic effect in psoriasis. The infiltrating T cells post-treatment have an activated cytotoxic phenotype, which may suggest a pathogenic role in the continuation and/or recurrence of psoriasis.     

Altered tissue expression pattern of cell adhesion molecules, ICAM-1, E-selectin and VCAM-1, in bullous pemphigoid during methotrexate therapy

Little is known about how eosinophils accumulate in bullous pemphigoid (BP) and why these cells rapidly disappear during immunosuppressive therapy. Eosinophils can produce cytokines such as IL-4, IL-5, IL-6, IL-10 and IL13, which can induce endothelial cells to express cellular adhesion molecules (CAMs) such as E-selectin, vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) necessary for the recruitment of eosinophils from the bloodstream to the skin. The present aim was to investigate the cellular expression of these three CAMs in serial biopsies before and during oral low-dose methotrexate therapy. Seventy-four biopsy specimens, 37 from active lesions and 37 from normal skin, were taken at different intervals from eight patients with bullous pemphigoid and stained immunohistochemically with specific monoclonal antibodies for these three CAMs. The expression and distribution of CAMs in the biopsies was evaluated and scored with light-microscopic examination. The basal keratinocytes in active lesions expressed ICAM-1. A strong VCAM-1 expression of endothelial cells and pericytes was correlated to a perivascular inflammatory cell infiltrate that also showed intense immunoreactivity to ICAM-1. Endothelial cell/pericytes also expressed E-selectin strongly in the BP patients before therapy. The expression of CAMs faded during therapy and, to the best of our knowledge, this has not been previously reported. Thus we suggest that the rapid reduction of tissue eosinophils may reflect the altered pattern of cell adhesion molecules during immunosuppressive therapy, which could explain the prompt clinical improvement seen in BP patients treated with methotrexate.     

Comparison of changes in endothelial adhesion molecule expression following UVB irradiation of skin and a human dermal microvascular cell line (HMEC-1)

We have assessed the pattern of dermal endothelial adhesion molecule expression following broadband UVB irradiation in vivo and in vitro. Skin biopsies were taken from 4 human volunteers at baseline and at 4, 8 and 24 h post-irradiation with 2.5 minimal erythema doses of UVB. Sections were stained immunohistochemically for E-selectin, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), CD31 and neutrophil elastase. The effect of direct UVB irradiation on E-selectin, ICAM-1 and VCAM-1 was examined in a human dermal microvascular endothelial cell line, HMEC-1. Cultured HMEC-1 were irradiated with 2.5–40 mJ/cm² of UVB, and assessed for adhesion molecule expression by immunofluorescence microscopy and fluorescence-activated cell sorter analysis. In vivo, E-selectin was minimally expressed on EC at baseline and was induced by 4 h following irradiation, P<0.01. ICAM-1 was moderately expressed at baseline and appeared mildly induced at 24 h, although this did not reach statistical significance. VCAM-1 was weakly expressed in unirradiated skin while CD31 was moderately expressed, but neither was induced by UVB irradiation. A significant neutrophilic infiltrate appeared by 8 h and was maximal at 24 h, P<0.05. Neutrophil infiltration correlated with E-selectin expression, r=0.96. In HMEC-1, ICAM-1 was upregulated at 24 h post-irradiation, with an increase in mean channel fluorescence from 100% at baseline to 145 (SD12)%> at 24 h, P<0.05. No change was seen in expression of E-selectin, VCAM-1 or CD31. These studies support the involvement of endothelial adhesion molecules E-selectin and ICAM-1 in UVB-induced inflammation. Whereas ICAM-1 is upregulated by direct irradiation of endothelial cells, E-selectin stimulation appears to be an indirect effect.     

ICAM-3 and E-selectin endothelial cell expression differentiate two phases of angiogenesis in infantile hemangiomas

Cellular adhesion molecules are newly identified mediators of angiogenesis. Infantile hemangiomas, characterized in the early stages by a proliferation of poorly differentiated vessels followed in the late stages by a vascular differentiation and regression of the tumor, represent an interesting model to study angiogenesis. We studied by immunohistochemistry the distribution of HLA-DR and three adhesion molecules ICAM-3, E-selectin and VCAM-1 on endothelial cells in different stages of vessel differentiation in infantile hemangiomas. We found high levels of ICAM-3 expression on proliferating vessels, while its expression was low or undetectable on well differentiated vessels. A different set of E-selectin antibodies showed a more heterogenous pattern of distribution and VCAM-1 antigens were found in both proliferating and differentiated vessels. HLA-DR expression on endothelial cells was inversely correlated to the vascular differentiation. Our results are consistent with the hypothesis that ICAM-3 plays a role in the early stages of vessel formation. Our results also suggest that variation of E-selectin and HLA-DR expression may be related either to vessel differentiation or may reflect the acquisition of an activated endothelial cell status.     

SLE患者外周血单一核细胞介导T淋巴细胞对血管内皮细胞黏附的实验研究

目的 探讨活动期SLE患者外周血单一核细胞(PBMC)是否介导了T淋巴细胞对血管内皮细胞的黏附.方法 体外培养人脐静脉内皮细胞(HUVEC),活动期SLE PBMC培养上清液预刺激48 h,通过RT-PCR检测PBMC培养上清液诱导HUVEC黏附分子的表达;划痕实验检测PBMC培养上清液诱导内皮细胞运动迁移能力的改变;HUVEC与T细胞共培养,研究PBMC培养上清液预刺激是否影响T细胞对HUVEC的黏附.结果 HUVEC经过活动期SLE PBMC培养上清刺激后,内皮细胞黏附分子ICAM-1mRNA,VCAM-1 mRNA和E-钙黏蛋白mRNA表达明显增加,而IL-17抗体能明显抑制黏附分子的表达.黏附分子表达增加介导了内皮细胞运动能力增加,介导T细胞对内皮细胞的黏附,而IL-17中和抗体能抑制T细胞对内皮细胞的黏附和抑制内皮细胞的运动迁移.结论 活动期SLE PBMC培养上清液可以通过诱导血管内皮细胞黏附分子表达,黏附T细胞,介导狼疮血管炎的发生.