Incorporation of fibronectin into matrix decreases TNF-induced increase in endothelial monolayer permeability.
Incorporation of fibronectin into matrix decreases TNF-induced increase in endothelial monolayer permeability.
复制标题
将纤连蛋白掺入基质可降低 TNF 诱导的内皮单层通透性增加。
DOI：
10.1152/ajplung.1993.265.2.l148
复制
发表时间：
1993
期刊：
影响因子：
--
通讯作者：
Saba,TM
中科院分区：
文献类型：
--
作者：
Wheatley,EM;McKeown-Longo,PJ;Vincent,PA;Saba,TM
血浆纤连蛋白是血液中的一种二聚体粘附蛋白，在肺中尤其是在血管损伤时掺入到内皮下和间质基质中。基质中的纤连蛋白被认为影响细胞-细胞相互作用和内皮细胞与富含胶原的细胞外基质的粘附。我们先前观察到，将纯化的可溶性人血浆纤连蛋白（hFn）加入培养的肺内皮细胞单层中，可减弱暴露于肿瘤坏死因子-α（TNF-α）的此类单层的蛋白渗透性增加。在目前的研究中，我们通过将hFn与人血浆中的另外两种纯化的粘附蛋白（即，玻连蛋白（Vn）和纤维蛋白原（Fg）。我们还通过将正常完整hFn与用N-乙基马来酰亚胺（NEM）烷基化的hFn或纯化的160/180-kDa hFn片段进行比较来确定基质掺入是否对于这种hFn介导的保护性应答是必不可少的，因为这些替代形式的纤连蛋白被认为表现出有限的掺入基质的能力。小牛肺动脉内皮（CPAE）单层（接种后3-4天）暴露于200 U/ml中浓度的人重组TNF-α 18 h，然后使用经内皮125 I标记的白蛋白清除率评估蛋白渗透性。二聚体hFn（600 μ g/ml）显著（P < 0.05）降低TNF诱导的内皮单层通透性增加。以等摩尔浓度加入到hFn中的Vn或Fg不能减弱内皮通透性。利用对hFn、人Vn或人Fg具有特异性的抗体的免疫荧光分析揭示了外源性hFn掺入细胞外基质中，但没有Vn或Fg的基质掺入。NEM处理的二聚体hFn以及纯化的160/180-kDa的hFn片段，其不能掺入基质中，也不能防止TNF诱导的蛋白质渗透性增加。因此，可溶性hFn减少TNF诱导的肺内皮细胞单层通透性增加的能力是特异性的，并依赖于其掺入细胞外基质。
Plasma fibronectin, a dimeric adhesive protein in blood, incorporates into the subendothelial and interstitial matrix in the lung especially during vascular injury. Fibronectin in the matrix is believed to influence cell-cell interaction and endothelial cell adhesion to the collagen-rich extracellular matrix. We previously observed that addition of purified soluble human plasma fibronectin (hFn) to cultured pulmonary endothelial monolayers attenuates the increase in protein permeability of such monolayers exposed to tumor necrosis factor-alpha (TNF-alpha). In the current study, we determined the specificity of this permeability response to fibronectin by comparing hFn to two other purified adhesive proteins in human plasma, i.e., vitronectin (Vn) and fibrinogen (Fg). We also determined whether matrix incorporation was essential for this hFn-mediated protective response by comparing normal intact hFn to either hFn alkylated with N-ethylmaleimide (NEM) or to purified 160/180-kDa hFn fragments, since these alternate forms of fibronectin are believed to exhibit limited ability to incorporate into matrix. Calf pulmonary artery endothelial (CPAE) monolayers (3-4 days postseeding) were exposed to human recombinant TNF-alpha for 18 h at a medium concentration of 200 U/ml followed by assessment of protein permeability using transendothelial 125I-labeled albumin clearance. Dimeric hFn (600 micrograms/ml) significantly (P < 0.05) reduced the TNF-induced increase in endothelial monolayer permeability. Vn or Fg, added at equal molar concentrations to the hFn, were unable to attenuate endothelial permeability. Immunofluorescent analysis utilizing antibodies specific to either hFn, human Vn, or human Fg revealed incorporation of the exogenous hFn into the extracellular matrix, but no matrix incorporation of Vn or Fg. Both NEM-treated dimeric hFn as well as purified 160/180-kDa fragments of hFn, which cannot incorporate into the matrix, were also unable to prevent the TNF-induced increase in protein permeability. Thus the ability for soluble hFn to reduce the TNF-induced increase in lung endothelial monolayer permeability was specific and dependent on its incorporation into the extracellular matrix.
登录
查看更多内容
使用高碘酸盐氧化检测碳水化合物表位特异性单克隆抗体。
影响因子：
2.2
作者：
Woodward,MP;YoungJr,WW;Bloodgood,RA
通讯作者：
Bloodgood,RA
影响因子：
2.9
作者：
KNECHT, DA;DIMOND, RL
通讯作者：
DIMOND, RL
中性粒细胞特异性抗原：免疫学和临床意义。
DOI：
--
发表时间：
1974
期刊：
Seminars in hematology (Print)
影响因子：
--
作者：
P. Lalezari;E. Radel
通讯作者：
E. Radel
CD15 单克隆抗体与人中性粒细胞表面含有磷酸酪氨酸的蛋白质发生反应。
DOI：
--
发表时间：
1988
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Skubitz,KM;Mendiola,JR;Collett,MS
通讯作者：
Collett,MS
使用“M Protocol”粒细胞单克隆抗体进行血清学、生化和细胞遗传学研究
影响因子：
3.1
作者：
P. Tetteroo;F. Visser;M. J. Bos;A. G. Kessel;J. Tromp;A. Borne
通讯作者：
A. Borne