Parker James C, Stevens Troy, Randall Jason, Weber David S, King Judy A
Dept. of Physiology, MSB 3074, College of Medicine, Univ. of South Alabama, Mobile, AL 36688, USA.
Am J Physiol Lung Cell Mol Physiol. 2006 Jul;291(1):L30-7. doi: 10.1152/ajplung.00317.2005.
Endothelial cells isolated from pulmonary arteries (RPAEC) and microcirculation (RPMVEC) of rat lungs were grown to confluence on porous filters and mounted on an Ussing-type chamber. Transmembrane pressure (deltaP) was controlled by the reservoir height, and the filtration rate corrected for surface area (J(v)/A) was measured by timing fluid movement in a calibrated micropipette. These parameters were used to calculate hydraulic conductance (Lp) by using linear regression of J(v)/A on deltaP. Mean Lp values for newly confluent RPAEC monolayers were 22 times higher than those for RPMVEC monolayers (28.6 +/- 5.6 vs. 1.30 +/- 0.50 x 10(-7) cm x s(-1) x cmH2O(-1); P < or = 0.01). After confluence was reached, electrical resistance and Lp remained stable in RPAEC but continued to change in RPMVEC with days in culture. Both phenotypes exhibited an initial time-dependent sealing response, but Lp also had an inverse relationship to deltaP in RPMVEC monolayers > or = 4 days postconfluence that was attributed to cell overgrowth rather than junctional length. In a comparison of the cadherin contents, E-cadherin was predominant in RPMVEC, but VE-cadherin was predominant in RPAEC. At a constant deltaP of 40-45 cmH2O for 2 h, J(v)/A increased 225% in RPAEC monolayers but did not change significantly in RPMVEC monolayers. Significant decreases in Lp were obtained after treatment with 5% albumin, GdCl3, or isoproterenol plus rolipram in both phenotypes. Thus lung microvascular endothelial cells exhibited a significantly lower Lp than conduit vessel endothelium, which would limit alveolar flooding relative to perivascular edema cuff formation during increased pulmonary vascular pressures.
从大鼠肺脏的肺动脉(RPAEC)和微循环(RPMVEC)中分离出的内皮细胞在多孔滤膜上生长至汇合,然后安装在尤斯廷型小室中。跨膜压力(ΔP)由储液器高度控制,通过校准微量移液器中液体移动的计时来测量校正表面积后的滤过率(J(v)/A)。这些参数用于通过对J(v)/A与ΔP进行线性回归来计算水力传导率(Lp)。新汇合的RPAEC单层的平均Lp值比RPMVEC单层高22倍(28.6±5.6对1.30±0.50×10⁻⁷ cm·s⁻¹·cmH₂O⁻¹;P≤0.01)。达到汇合后,RPAEC中的电阻和Lp保持稳定,但RPMVEC中的电阻和Lp在培养天数增加时继续变化。两种表型都表现出初始的时间依赖性封闭反应,但在汇合后≥4天的RPMVEC单层中,Lp也与ΔP呈负相关,这归因于细胞过度生长而非连接长度。在钙黏蛋白含量的比较中,E-钙黏蛋白在RPMVEC中占主导,但VE-钙黏蛋白在RPAEC中占主导。在40 - 45 cmH₂O的恒定ΔP下处理2小时,RPAEC单层中的J(v)/A增加了225%,但RPMVEC单层中没有显著变化。用5%白蛋白、GdCl₃或异丙肾上腺素加咯利普兰处理后,两种表型的Lp均显著降低。因此,肺微血管内皮细胞的Lp明显低于管道血管内皮细胞,这在肺血管压力升高期间相对于血管周围水肿袖套形成会限制肺泡积水。