山莨菪碱对内毒素致肺微血管内皮细胞骨架变化的影响

目的观察内毒素（ＬＰＳ）对肺微血管内皮细胞（ＰＭＶＥＣ）单层通透性和肌动蛋白骨架的影响，探讨与β受体和Ｇｓα蛋白变化的关系及山莨菪碱的干预作用。方法体外培养大鼠ＰＭＶＥＣ，采用放射性配基结合分析法和流式细胞仪技术。结果ＬＰＳ在体外可诱导ＰＭＶＥＣ单层通透性增高和中央Ｆ－肌动蛋白发生解聚、密度明显减低，同时出现β受体下调及Ｇｓα蛋白水平降低；山莨菪碱对上述变化有明显抑制作用。结论ＬＰＳ可直接导致ＰＭＶＥＣ单层通透性增高，并与Ｆ－肌动蛋白重排相关。山莨菪碱可能通过影响ＰＭＶＥＣ膜β受体和Ｇｓα蛋白两个环节，参与调控ＰＭＶＥＣ的肌动蛋白骨架变化，减轻ＬＰＳ所致的大鼠ＰＭＶＥＣ单层通透性增高。

Effects of anisodamine on the change of cytoskeleton of rat pulmonary microvascular endothelial cells induced by endotoxin

AIM　Observing the influence of lipopolysaccharide (LPS) on changes of the permeability and actin cytoskeleton of rat pulmonary microvascular endothelial cells (PMVEC) monolayer, we explore their relationship with β-adrenoceptor(β-AR) and G protein subunit(Gsα),and the interfering action of anisodamine. METHODS　PMVEC was isolated and cultured from Wistar rat in vitro, radioligand binding assay and flow cytometer were used. RESULTS　After incubation of LPS in vitro, the central F-actin of PMVEC depolymerized and its density decreased obviously ,while the permeability of PMVEC monolyer increased significantly. Meanwhile it was found that LPS can induce the down regulation of β-AR and Gsα protein level. Anisodamine can inhibit these changes above. CONCLUSION　LPS can directly cause the increase of permeability of PMVEC monolayer which relates to the depolymerization of central F-actin in PMVEC. Anisodamine may take part in regulating actin cytoskeleton of PMVEC and attenuate LPS-induced the increase of permeability of PMVEC monolayer by influencing two links of both β-AR and Gsα protein.