Résumé
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.
Résumé
AIM: To study the effect of lipopolysaccharide(LPS) on cytosolic Ca 2+ and Gq protein in pulmonary microvascular endothelial cells of rats (RPMVECS) and the interferring action of anisodamine.METHODS: RPMVECS of Wistar rats were isolated, cultured and identified. The concentration of cytosolic Ca 2+ and Gq protein were measured by Fura-2/AM and flow cytometry, respectively.RESULTS: (1) Compared with the control group, the level of cytosolic Ca 2+ increased and the level of Gq protein decreased significantly at 30 min and 90 min after stimulation of LPS;(2) Increase in cytosolic Ca 2+ and decrease in Gq protein in RPMVECS were inhibited by anisodamine. CONCLUSION: LPS induced the increase in cytosolic Ca 2+ and the decrease in Gq protein in RPMVEC, which was inhibited by anisodamine.
Résumé
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 ?-adrenoceptor(?ＡＲ) and G protein subunit(Gs?), and the interfering action of anisodamine. METHODS PMVEC was isolated and cultured from Wistar rat in vitro, radi- oligand 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 5-AR and Gsa protein level. Anisodamine can in- hibit 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 monolaver by influencing two links of both ?-AR and Gsa protein