复方丹参对高转移性人肺癌细胞与血管内皮细胞黏附及黏附分子表达的影响

目的　探讨复方丹参抗肿瘤转移机理。方法　应用共聚焦显微镜和流式细胞仪荧光标记法,体外研究高转移性肺癌细胞(PG细胞)与血管内皮细胞的黏附性、黏附分子表达以及复方丹参抗肿瘤转移作用。结果　复方丹参可明显抑制PG细胞表面CD4 4 ,CD5 4的表达。复方丹参对PG细胞与激活和静息血管内皮细胞的黏附性也具有明显的抑制作用,并可抑制黏附分子CD4 4 ,CD4 5的表达,呈剂量依赖关系。结论　复方丹参具有抗肿瘤转移作用,抑制肿瘤细胞与内皮细胞黏附及黏附分子表达可能是复方丹参抗肿瘤转移的机制之一。     

新加沙参麦冬煎剂抑制肿瘤转移及其作用机制的实验研究

目的 探讨新加沙参麦冬煎剂防治肿瘤转移的机制。方法结合动物模型、免疫组化、细胞分子生物学技术，观察新加沙参麦冬煎剂对荷瘤小鼠抑瘤率、转移抑制率、生命延长率的影响，检测实验小鼠皮下移植瘤血管内等细胞生长因子（VEGF）、血管内皮细胞第Ⅷ因子（CD34）、黏附因子（CD44V6）、基质金属蛋白酶2（MMP2）、基质金属蛋白酶抑制酶（TIMP2）的表达情况。结果新加沙参麦冬煎剂治疗组的瘤重抑制率、肺转移抑制率、生命延长率分别为37.3%，58.3% 和20.8%；皮下移植瘤CD44V6 、VEGF表达、微血管密度（MVD）明显低于空白组（P＜0.01），TIMP2表达记分明显高出其他组（P＜0.01）。小鼠肺转移灶数与VEGF 、CD44V6、MVD呈线性相关，相关系数分别为0.490，0.398，0.455。结论①新加沙参麦冬煎剂对小鼠LA795高转移肺腺癌模型有较好抑制转移、抑制肿瘤生长和延长荷瘤小鼠生存时间的作用。②新加沙参麦冬煎剂可通过调控肿瘤转移过程中黏附、基质降解、血管生成相关分子的表达，多途径、多靶点防治肿瘤的转移。     

食管癌放化疗治疗对淋巴细胞黏附分子表达的作用及临床意义

目的：探讨放化疗治疗对食管癌患者外周血淋巴细胞CD44V6、CD54表达的作用及临床意义。方法：采用流式细胞术对食管癌及正常成人外周血淋巴细胞CD44V6、CD54的表达水平进行检测,并比较不同病理类型、分期及放化疗前后淋巴细胞黏附分子表达水平的变化。结果：食管癌患者外周血CD44V6、CD54的表达水平显著高于正常对照组（P〈0．01）,其表达水平与肿瘤病理类型和临床分期密切相关,放化疗后患者的黏附分子表达水平明显下降。结论：食管癌患者外周血淋巴细胞CD44V6、CD54的高表达与肿瘤分期及转移密切相关,经治疗后其表达水平明显下降．检测其表达的动态变化可以为评估肿瘤发展及疗效判断提供依据。     

山楂叶含药血清对高脂血清诱导单核细胞与血管内皮细胞黏附能力的影响

目的研究山楂叶含药血清对高脂血清诱导的单核细胞与血管内皮细胞黏附能力及黏附分子表达的影响,并探讨其作用机制。方法采用虎红染色法检测药物血清对高脂诱导的人脐静脉内皮细胞(human umbilical vascular endothelial cells,HUVEC)和单核细胞株THP-1黏附的作用;用细胞ELISA法检测HUVEC表面细胞间黏附分子-1(intercelluar adhesion molecule-1,ICAM-1)、血管细胞黏附分子-1(vascular cellular adhesion molecule-1,VCAM-1)、P-选择素(P-selectin)的表达。结果山楂叶能抑制高脂诱导的HUVEC与THP-1的黏附,并下调HUVEC表面ICAM-1、VCAM-1、P-se-lectin的表达。结论山楂叶防治高脂血症的机制之一可能是下调血管内皮细胞黏附分子的表达,减少单核细胞与内皮细胞的黏附,从而抑制单核细胞迁移至血管内膜形成泡沫细胞而实现。     

肺瘤平Ⅱ号对肺癌患者血浆CD44v6及CD49表达的影响*

目的：研究中药复方制剂肺瘤平Ⅱ号对非小细胞肺癌患者血浆CD44 v6和CD49表达的影响，以及血浆CD44 v6和CD49表达与淋巴结转移的关系，从分子水平探讨了肺瘤平Ⅱ号抗肿瘤的作用机制。方法：中晚期非小细胞肺癌患者60例，随机分为对照组和试验组各30例，并选择健康人组10人。对照组予化疗，3个周期为1疗程。试验组在化疗的基础上给予肺瘤平Ⅱ号膏口服，每次20g，tid，3个月为1疗程。治疗前后以流武细胞术检测患者血浆中CD44 v6及CD49含量，观察淋巴结转移情况，两治疗组各随机抽取10例，同健康人组进行对照。结果：试验组CD44 v6及CD49较治疗前含量显著降低（P〈0．01或P〈0．05）；而对照组治疗前后变化不明显（P〉0．05）。淋巴结转移阳性患者CD44 v6或CD49血浆含量较淋巴结阴性患者为高（P〈0．05）。临床检测结果与动物实验基本一致。结论：中药复方制剂肺瘤平Ⅱ号通过抑制肿瘤细胞CD44 v6和CD49的表达，发挥抗肿瘤作用。     

志苓胶囊对小细胞肺癌细胞株NCI-H446细胞凋亡及h-TERT、CD44表达的影响

目的研究志苓胶囊(抗癌复方Ⅱ号)对人小细胞肺癌细胞株NCI-H446诱导凋亡作用机制以及对细胞黏附分子表达的影响。方法将志苓胶囊按其不同的中西药成份配制成相应的中药组、西药组和复方组,与NCI-H446共培养后,倒置显微镜观察、Hochest33258荧光染色法检测凋亡细胞,RT-PCR、Western blot、流式细胞仪分别检测bcl-2、bax、hTERT基因mRNA和相应蛋白表达水平变化以及黏附分子CD44的表达变化。结果NCI-H446经不同的药物组处理后,倒置显微镜、Hochest33258荧光染色可观察到细胞凋亡形态学改变,bcl-2基因mRNA和蛋白表达水平均有不同程度下降,bax基因表达水平则有不同程度增强,复方组改变最明显,各药物组与对照组比较差异具有显著性(P<0.01)。中药组和西药组hTERT基因和蛋白表达水平未见明显改变,复方组表达水平下调,与对照组比较差异具有显著性(P<0.01)。各药物组CD44表达水平降低,其中复方组降低水平最明显(P<0.01)。结论志苓胶囊可能通过下调NCI-H446细胞bcl-2、hTERT基因和CD44分子表达,增强bax基因表达,从而诱导NCI-H446细胞凋亡,抑制细胞黏附、转移。     

罗仙子提取物对单核细胞与血管内皮细胞黏附的影响

目的：考察罗仙子提取物对单核细胞与血管内皮细胞黏附的影响。方法：制备相对分子质量30000以下罗仙子提取物;考察罗仙子提取物对血管内皮细胞EA．hy926活力的影响;肿瘤坏死因子（TNF-α）诱导建立单核细胞THP-1和EA．hy926黏附模型,采用虎红染色,考察不同浓度罗仙子提取物对EA．hy926与THP-1黏附性的影响;采用ELISA考察罗仙子提取物对TNF-α诱导的EA．hy926分泌单核细胞趋化蛋白-1（MCP-1）和血管细胞黏附分子-1（VCAM-1）的影响。结果：罗仙子提取物（浓度≤4×102μg／mL时）对EA．hy926未见明显的增殖或抑制作用（P〉0．05）;罗仙子提取物（浓度≥6．25μg／mL时）可浓度依赖性降低两细胞之间的黏附作用;罗仙子提取物各浓度均能显著地降低TNF-α诱导的MCP-1表达（P〈0．01）,罗仙子提取物（浓度≥50μg／mL时）可显著地降低TNF-α诱导的VcAM-1的表达（P〈0．01）。结论：罗仙子提取物可通过抑制TNF-α诱导的内皮细胞分泌MCP-1和VCAM-1,阻滞单核细胞与血管内皮细胞之间的黏附作用,从而发挥抗动脉粥样硬化作用。     

茶多酚对肺癌PG细胞生长的调控研究

目的：探讨茶多酚对人PG细胞生长的影响。方法：采用MTT法体外观察不同浓度茶多酚对PG细胞的杀伤作用；应用激光共聚焦显微镜和流式细胞仪分析，检测用药后PG细胞内Ca^2 浓度，CD44V6表达和细胞增殖周期分布的变化。结果：3个浓度的茶多酚对PG2细胞的有杀伤作用，呈剂量依赖分析。细胞增殖受到明显抑制，使细胞阻滞于G0／G1期，不能进入S期及G2／M期，细胞增殖指数明显下降。细胞内Ca^2 浓度与对照组相比，逐渐上升；CD44V6表达水平则呈逐渐下降。结论：茶多酚对PG细胞的生长具有抑制作用，其作用机制与改变细胞内Ca^2 浓度和CD44V6表达有关。     

黏附分子CD44与恶性瘤关系的研究进展

在细胞生长和分化过程中,变异性剪接是许多基因表达过程的一部分。CD44作为多功能黏附分子,有10个变异外显子能发生选择性拼接,从而产生CD44s和CD44v,其中CD44v主要表达在转移性肿瘤细胞表面。在许多恶性瘤组织中可见到不同CD44v分子的表达。此文综述了CD44分子与各种肿瘤侵袭转移的相关性。     

低分子量肝素对白细胞黏附和黏附分子表达的影响

目的研究低分子量肝素对白细胞与血管内皮细胞体内外黏附和细胞间黏附分子 1(ICAM 1)表达的影响。方法倒置显微镜观察在致敏豚鼠肠系膜微循环中白细胞的滚动和黏附情况 ,体外观察大鼠中性粒细胞对培养的脐静脉内皮细胞的黏附情况 ,流式细胞仪测定ICAM 1的表达。结果低分子量肝素 2 0、4 0u kg对白细胞黏附有明显抑制作用 ;0 .5、1.0u ml(终浓度 )对中性粒细胞体外黏附有明显抑制作用 ;0 .12 5、0 .2 5、0 .5、1.0u ml对脐静脉内皮ICAM 1的表达有明显抑制作用。结论低分子量肝素有明显抑制白细胞黏附及降低ICAM 1表达的作用。     

商陆皂苷甲对细胞间粘附的影响商陆皂苷甲对细胞间粘附的影响

目的研究商陆皂苷甲(EsA)对细胞间粘附的影响，并观察粘附分子ICAM-1和CD18 mRNA的表达水平。方法用血细胞计数仪计数的方法考察在脂多糖（LPS）诱导下，EsA对中性粒细胞与内皮细胞间粘附的影响。用逆转录聚合酶链反应（RT-PCR）的方法测定内皮细胞ICAM-1 mRNA和中性粒细胞CD18 mRNA的表达水平。结果EsA在3～12×10^-6　μmol·L^-1能降低LPS作用下中性粒细胞与内皮细胞间高水平的粘附率，且能降低LPS诱导下内皮细胞ICAM-1 mRNA和中性粒细胞CD18 mRNA的表达。结论EsA能抑制LPS作用下中性粒细胞与内皮细胞间的粘附可能是其抗炎机制之一，降低粘附分子的表达可能是其影响细胞间粘附的重要作用机制。     

Astilbin inhibits the adhesion of T lymphocytes via decreasing TNF-alpha and its associated MMP-9 activity and CD44 expression

Our previous study has revealed that astilbin, a flavonoid isolated from the rhizome of Smilax glabra, improves an immunological liver injury and its mechanism includes an inhibition of the lymphocyte adhesion. The present study further examined the anti-adhesive activity in various assays by using human leukemia T cell line Jurkat cells. We found that astilbin inhibited the adhesion of Con A or PMA-activated Jurkat cells to fibronectin, type IV collagen, hyaluronic acid, and [corrected] ECV-304 cells. Astilbin inhibited the adhesion of Jurkat cells to PMA-activated but not non-activated ECV-304 cells without any influence on the survival of the ECV-304 cells and Jurkat cells. Astilbin also inhibited the CD44 expression and TNF-alpha production in Jurkat cells. In the co-culture assay between Jurkat cells and ECV-304 cells, the MMP-9 secretion from Jurkat cells was inhibited after astilbin-treatment, while the exogenous TNF-alpha increased the MMP-9 secretion in a dose-dependent manner. These findings suggest that the inhibition of T lymphocyte adhesion by astilbin may be related to the reduction of the CD44 expression and TNF-alpha production in the cells, which may further cause a decreased MMP-9 secretion.     

粘附分子CD54、CD44在苦瓜蛋白诱导K562细胞凋亡中的作用

目的观察苦瓜蛋白诱导K562细胞凋亡时细胞粘附分子(CD54、CD44)表达水平的变化,探讨粘附分子在苦瓜蛋白诱导K562细胞凋亡中的作用。方法以一定浓度的苦瓜蛋白处理K562细胞;CCK-8试剂盒检测其对细胞生长的影响;流式细细术(An-nexinⅤ)、透射电镜等方法检测细胞凋亡;同时应用流式细细术检测粘附分子(CD54、CD44)蛋白表达水平的变化。结果苦瓜蛋白对K562细胞生长具有明显的抑制作用;流式细胞术及形态学观察(电镜)证实一定作用浓度的苦瓜蛋白可诱导K562细胞发生明显的细胞凋亡;苦瓜蛋白处理组K562细胞中CD54、CD44表达分别为18.62%和1.32%,对照组分别为0.25%和0.17%,分别上调了18.37%、1.15%。结论苦瓜蛋白引起K562细胞凋亡的过程中,粘附分子CD54表达上调;CD54表达上调可能是苦瓜蛋白诱导肿瘤细胞凋亡的机制之一,其机制可能涉及细胞粘附依赖性细胞凋亡。     

Anti-inflammatory mechanisms of apigenin: inhibition of cyclooxygenase-2 expression,adhesion of monocytes to human umbilical vein endothelial cells,and expression of cellular adhesion molecules

The aim of this study was to clarify the anti-inflammatory mechanism of apigenin. Apigenin inhibited the collagenase activity involved in rheumatoid arthritis (RA) and suppressed lipopolysaccharide (LPS)-induced nitric oxide (NO) production in a dose dependent manner in RAW 264.7 macrophage cells. Pretreatment with apigenin also attenuated LPS-induced cyclooxygenase-2 (COX-2) expression. In addition, apigenin profoundly reduced the tumor necrosis factor-alpha (TNF-alpha)-induced adhesion of monocytes to HUVEC monolayer. Apigenin significantly suppressed the TNF-alpha-stimulated upregulation of vascular cellular adhesion molecule-1 (VCAM-1)-, intracellular adhesion molecule-1 (ICAM-1)-, and E-selectin-mRNA to the basal levels. Taken together, these results suggest that apigenin has significant anti-inflammatory activity that involves blocking NO-mediated COX-2 expression and monocyte adherence. These results further suggest that apigenin may be useful for therapeutic management of inflammatory diseases.     

CXCL12/CXCR7对胃癌生长、粘附、侵袭的影响

目的：运用免疫组化方法研究CXCL12/CXCR7在胃癌组织中的表达,并选取与粘附、侵袭有关的几个细胞因子进行相关性研究,揭示CXCR7与胃癌生长、粘附及侵袭的关系。方法：收集160例胃腺癌病人的癌组织和30例正常胃组织,采用免疫组化方法,检测CXCL12/CXCR7、Survivin、CD44v6、MMP-3在胃癌和正常组织中的表达,并用统计学方法分析其相关性。结果：CXCL12/CXCR7在胃癌组织中的表达明显高于对照组正常胃组织（P=0.000）,并且其在胃癌中的表达与胃癌的大小、浸润深度、是否有淋巴结转移及临床分期有关（P﹤0.05）。在胃癌组织中CXCR7表达与Survivin、CD44v6及MMP-3呈正相关。结论：CXCR7通过抗凋亡(Survivin)参与胃癌的生长,在胃癌的粘附及侵袭过程中通过CD44V6及MMP-3发挥作用,并可能参与胃癌的淋巴结转移。     

The inhibition of neutrophil-endothelial cell adhesion by hyaluronan independent of CD44

Objective: To study the effect of hyaluronan on cell adhesion and recruitment both in vitro and in vivo, since hyaluronan both inhibits restenosis and is anti-inflammatory. When administered to animals undergoing angioplasty the recruitment of cells into the restenotic plaque is inhibited, as well as into inflammatory lesions. The recent discovery that ICAM-1 binds hyaluronan and exhibits the B(X(7))B HA binding motif, led us also to investigate whether cell adhesion could be modulated by hyaluronan. Materials and methods: Human neutrophils were adhered to human umbilical vein (HUVEC) or Ea.hy.926 HUVEC cells stimulated with phorbol myristate acetate (PMA) or tumour necrosis factor (TNFα). Neutrophil binding in vivo utilized FMLP-stimulated hamster cheek pouch post-capillary venules. Results: Hyaluronan inhibited human neutrophil adhesion to both PMA and TNFα-stimulated HUVEC. Ea.hy.926 human immortal HUVECs expressed ICAM-1 in response to TNFα and PMA. E-selectin was also upregulated by 6 h with TNFα but not significantly with PMA. TNFα induced CD44 expression within 4 h, but PMA not significantly up to 6 h. However, specific binding of [¹²⁵I]hyaluronan to Ea.hy.926 cells was increased by PMA-stimulation at 4 h. Neutrophil adhesion to PMA-stimulated Ea.hy.926 HUVECs was inhibited in a concentration dependent fashion by both anti-ICAM-1 and hyaluronan (1 ng/ml-10 μg/ml) at 4 h. At 1 mg/ml adhesion was stimulated by hyaluronan. Hyaluronan had no effect on neutrophil adhesion to resting Ea.hy.926 cells. Hyaluronan (25 mg/kg, i.v.) inhibited cell adhesion to FMLP-stimulated post capillary venules of the hamster cheek pouch, whilst leaving cell rolling unaffected. Conclusions: These results show that hyaluronan, at concentrations below those where intramolecular associations occur, binds selectively to stimulated endothelial cells and inhibits neutrophil adhesion in vitro and in vivo via a mechanism which may involve molecules other than CD44, such as ICAM-1. Deceased.     

Nitric oxide regulates human eosinophil adhesion mechanisms in vitro by changing integrin expression and activity on the eosinophil cell surface

1. The nitric oxide synthase (NOS) inhibitor, N(omega)-nitro-L-arginine methyl ester (L-NAME), inhibits both rat and human eosinophil chemotaxis in vitro. Here, the role of nitric oxide (NO) in human eosinophil cell surface integrin expression and function was investigated. 2. Human peripheral blood eosinophils were treated with L-NAME (0.01 - 1.0 mM) and their adhesion to human fibronectin and serum observed. Adhesion of cells to fibronectin and serum increased by 24.0+/-4.6 and 43.8+/-4.7%, respectively, when eosinophils were treated with 1.0 mM L-NAME. Increased adhesion by L-NAME could be abolished when cells were co-incubated with VLA-4- and Mac-1-specific monoclonal antibodies (mAbs). 3. The NO donor, sodium nitroprusside (2.5 mM), significantly inhibited eosinophil adhesion to fibronectin and serum by 34.3+/-4.5 and 45.2+/-5.6%, respectively. This inhibition was accompanied by a 4 fold increase in the levels of intracellular cyclic GMP. 4. Flow cytometrical analysis demonstrated that L-NAME induced an increased expression of CD11b (Mac-1) on the eosinophil cell surface of 36.3+/-7.4%. L-NAME had no effect upon CD49d (VLA-4) expression. 5. Treatment of human eosinophils, in vitro, with H-[1,2,4] oxadiazolo quinoxalin-1-one (ODQ) (0.1 mM), an inhibitor of soluble guanylate cyclase, also significantly increased eosinophil adhesion to fibronectin and serum by 73.5+/-17.9 and 91.7+/-12.9%, respectively. This increase in adhesion could also be inhibited by co-incubation with the Mac-1 and VLA-4-specific mAbs. 6. In conclusion, results indicate that NO, via a cyclic GMP-dependent mechanism, inhibits the adhesion of human eosinophils to the extracellular matrix (ECM). This inhibition is accompanied by a decrease in the expression and function of the eosinophil's adhesion molecules, in particular, the expression of the Mac-1 integrin and the function of the VLA-4 integrin.     

Effect of Linomide on adhesion molecules, TNF-alpha, nitrogen oxide, and cell adhesion

Linomide (quinoline-3-carboxamide) is an immunomodulator with anti-inflammatory effects in rodents with autoimmune diseases. Its mode of action still remains to be elucidated. We hypothesized that an investigation of T cell interactions with the extracellular matrix (ECM), composed of glycoproteins such as fibronectin (FN) and laminin (LN), might provide better understanding of their in vivo mode of action in extravascular inflammatory sites. We examined the effect of Linomide on T cell adhesion to intact ECM, and separately to LN, and FN, and on the release and production of tumor necrosis factor (TNFalpha) and nitrogen oxide (NO) in relation to adhesive molecules in non-obese diabetic (NOD) female spleen cells, focusing on intracellular adhesion molecule-1 (ICAM-1) and CD44. NOD female mice that developed spontaneous autoimmune insulitis, which destroys pancreatic islets and subsequently leads to insulin-deficient diabetes mellitus, were studied. Linomide, given in the drinking water or added to tissue cultures in vitro, inhibited the beta1 integrin-mediated adhesion of T cells to ECM, FN and LN, as well as the production and release of TNFalpha and NO, which play a major role in the induction and propagation of T cell-mediated insulitis. In addition, exposure of T cells to Linomide resulted in increased expression of CD44 and ICAM-1 molecules on spleen cells of Linomide-treated mice; such an increase in adhesion molecule expression may lead to more effective arrest of T cell migration in vivo. The regulation of T-cell adhesion, adhesion receptor expression, and inhibition of TNFalpha and NO secretion by Linomide may explain its beneficial role and provide a new tool for suppressing self-reactive T cell-dependent autoimmune diseases.