山莨菪碱对血小板活化因子致肺微血管内皮细胞形态和骨架变化的影响

Effects of anisodamine on the injury of rat pulmonary microvascular endothelial cells induced by platelet-activating factor

目的 观察血小板活化因子 (PAF)对培养肺微血管内皮细胞形态、单层通透性、肌动蛋白骨架的影响及山莨菪碱的干预作用。方法 HE染色观察PAF对体外培养大鼠肺微血管内皮细胞 (RPMVEC)的形态影响 ;用针头式滤器观察PAF对RPMVEC单层通透性的影响 ;用免疫细胞化学的方法观察PAF对RPMVECF 肌动蛋白 (F actin)分布的影响 ;并观察山莨菪碱干预后的变化。结果 PAF浓度大于0 1mg/L作用 4 8h内观察到细胞脱落和破裂。 10mg/LPAF 12 0min内可使RPMVEC单层通透性增高、F actin解聚 ,山莨菪碱可抑制上述变化。结论 PAF引起RPMVEC脱落和破裂呈时间和剂量依赖性 ;PAF引起内皮单层通透性的增高的机制与F actin解聚密切相关 ;山莨菪碱可抑制PAF引起RPMVEC细胞脱落、单层通透性增高和F actin解聚 。

Objective To investigate the mechanism of platelet-activating factor(PAF) induced injury on the cultured rat pulmonary microvascular endothelial cells (RPMVEC),and the interfering action of anisodamine. Methods RPMVEC were isolated and cultured from Wistar rat in vitro,the effects of PAF on morphology,monolayer permeability; F-actin of RPMVEC were observed. F-actin expression was evaluated by immunocytochemistry. Results PAF in the concentration higher than 0 1 mg/L induced detachment and rupture of RPMVEC within 48 h. 10 mg/L of PAF increased permeability of RPMVEC monolayer and depolymerization of F-actin within 120 min. Anisodamine inhibited the effects of PAF. Conclusion The detachment and rupture of RPMVEC induced by PAF depends on the exposed concentration and time. The increased permeability of RPMVEC monolayer induced by PAF is significantly correlated with the depolymerization of F-actin. The increased permeability of RPMVEC monolayer and depolymerization of F-actin induced by PAF are markedly inhibited by anisodamine,which exerts protective action on RPMVEC injury induced by PAF.