Suttorp N, Polley M, Seybold J, Schnittler H, Seeger W, Grimminger F, Aktories K
Department of Internal Medicine, Justus Liebig-University, Giessen, Germany.
J Clin Invest. 1991 May;87(5):1575-84. doi: 10.1172/JCI115171.
The endothelial cytoskeleton is believed to play an important role in the regulation of endothelial permeability. We used botulinum C2 toxin to perturb cellular actin and determined its effect on the permeability of endothelial cell monolayers derived from porcine pulmonary arteries. The substrate for botulinum C2 toxin is nonmuscle monomeric actin which becomes ADP-ribosylated. This modified actin cannot participate in actin polymerization and, in addition, acts as a capping protein. Exposure of endothelial cell monolayers to botulinum C2 toxin resulted in a dose- (3-100 ng/ml) and time-dependent (30-120 min) increase in the hydraulic conductivity and decrease in the selectivity of the cell monolayers. The effects of C2 toxin were accompanied by a time- and dose-dependent increase in ADP-ribosylatin of G-actin. G-Actin content increased and F-actin content decreased time- and dose-dependently in C2 toxin-treated endothelial cells. Phalloidin which stabilizes filamentous actin prevented the effects of botulinum C2 toxin on endothelial permeability. Botulinum C2 toxin induced interendothelial gaps. The effects occurred in the absence of overt cell damage and were not reversible within 2 h. The data suggest that the endothelial microfilament system is important for the regulation of endothelial permeability.
内皮细胞骨架被认为在调节内皮通透性方面发挥重要作用。我们使用肉毒杆菌C2毒素干扰细胞肌动蛋白，并确定其对源自猪肺动脉的内皮细胞单层通透性的影响。肉毒杆菌C2毒素的底物是非肌肉单体肌动蛋白，其会发生ADP-核糖基化。这种修饰的肌动蛋白不能参与肌动蛋白聚合，此外，还作为一种封端蛋白。将内皮细胞单层暴露于肉毒杆菌C2毒素会导致水力传导率呈剂量（3 - 100 ng/ml）和时间依赖性（30 - 120分钟）增加，以及细胞单层选择性降低。C2毒素的作用伴随着G-肌动蛋白的ADP-核糖基化呈时间和剂量依赖性增加。在C2毒素处理的内皮细胞中，G-肌动蛋白含量随时间和剂量依赖性增加，F-肌动蛋白含量随时间和剂量依赖性降低。稳定丝状肌动蛋白的鬼笔环肽可阻止肉毒杆菌C2毒素对内皮通透性的影响。肉毒杆菌C2毒素诱导内皮细胞间出现间隙。这些作用在没有明显细胞损伤的情况下发生，并且在2小时内不可逆。数据表明内皮微丝系统对调节内皮通透性很重要。
