Moy Alan B, Blackwell Ken, Wu Mack H, Granger Harris J
Cellular Engineering Technologies, Inc., 2501 Crosspark Rd., Ste. B105, Coralville, IA 52241, USA.
Am J Physiol Heart Circ Physiol. 2006 Nov;291(5):H2126-35. doi: 10.1152/ajpheart.00185.2006. Epub 2006 May 5.
We report functional differences in constitutive and agonist-mediated endothelial barrier function between cultured primary and Clonetics human umbilical vein endothelial cells (pHUVEC and cHUVEC) grown in soluble growth factors and heparin. Basal transendothelial resistance (TER) was much lower in pHUVEC than in cHUVEC grown in medium supplemented with growth factors, such as basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), and human epithelial growth factor (EGF), and heparin. On the basis of a numerical model of TER, the increased basal TER in cHUVEC was due to effects on cell-matrix adhesion and membrane capacitance. Heparin and bFGF increased constitutive TER in cultured pHUVEC, and heparin mediated additional increases in constitutive TER in pHUVEC supplemented with bFGF. EGF attenuated bFGF-mediated increases in TER. On the basis of the numerical model, in contrast to cHUVEC, heparin and bFGF augmented TER through effects on cell-cell adhesion and membrane capacitance in pHUVEC. Thrombin mediated quantitatively greater amplitude and a more sustained decline in TER in cultured cHUVEC than pHUVEC. Thrombin-mediated barrier dysfunction was attenuated in pHUVEC conditioned in EGF in the presence or absence of heparin. Thrombin-mediated barrier dysfunction was also attenuated when monolayers were exposed to low concentrations of heparin and further attenuated in the presence of bFGF. cAMP stimulation mediated differential attenuation of thrombin-mediated barrier dysfunction between pHUVEC and cHUVEC. VEGF displayed differential effects in TER in serum-free medium. Taken together, these data demonstrate marked differential regulation of constitutive and agonist-mediated endothelial barrier function in response to mitogens and heparin stimulation.
我们报告了在可溶性生长因子和肝素中培养的原代和Clonetics人脐静脉内皮细胞（pHUVEC和cHUVEC）之间，组成型和激动剂介导的内皮屏障功能的功能差异。在补充有生长因子（如碱性成纤维细胞生长因子（bFGF）、血管内皮生长因子（VEGF）和人表皮生长因子（EGF））和肝素的培养基中生长的pHUVEC的基础跨内皮电阻（TER）远低于cHUVEC。基于TER的数值模型，cHUVEC中基础TER的增加是由于对细胞-基质粘附和膜电容的影响。肝素和bFGF增加了培养的pHUVEC中的组成型TER，并且肝素介导了补充有bFGF的pHUVEC中组成型TER的额外增加。EGF减弱了bFGF介导的TER增加。与cHUVEC相反，基于数值模型，肝素和bFGF通过对pHUVEC中细胞-细胞粘附和膜电容的影响增强了TER。凝血酶介导的培养的cHUVEC中TER的下降幅度在数量上大于pHUVEC，且更持久。在有或没有肝素的情况下，在EGF中预处理的pHUVEC中，凝血酶介导的屏障功能障碍减弱。当单层暴露于低浓度肝素时，凝血酶介导的屏障功能障碍也减弱，并且在存在bFGF的情况下进一步减弱。cAMP刺激介导了pHUVEC和cHUVEC之间凝血酶介导的屏障功能障碍的差异减弱。VEGF在无血清培养基中对TER显示出不同的影响。综上所述，这些数据表明，在有丝分裂原和肝素刺激下，组成型和激动剂介导的内皮屏障功能存在明显的差异调节。
