摘要Objective:It has been documented that ezrin/radixin/moesin (ERM) phosphorylation by the p38 mitogen-activated protein kinase (MAPK),Rho/ROCK,and protein kinase C (PKC) pathways leads to filamentous actin (F-actin) reorganization and microvascular endothelial cell hyperpermeability.In this study,we investigated the effects of Xijiao Dihuang Decoction combined with Yinqiao Powder (XDY) on influenza virus (Ⅳ)-induced F-actin restructuring and ERM phosphorylation regulated by the Rho/Rho kinase 1 (ROCK),p38MAPK,and PKC signaling pathways in pulmonary microvascular endothelial cells (PMVECs).Methods:Serum containing XDY (XDY-CS;13.8 g/kg) was acquired using standard protocols for serum pharmacology.Primary PMVECs were obtained from male Wistar rats and cultured.After adsorption of Ⅳ A (multiplicity of infection,0.01) for 1 h,medium with 20％ XDY-CS was added to the PMVECs.The distributions of F-actin and phosphorylated ERM were determined by confocal microscopy,and F-actin expression was measured by flow cytometry.The expression levels of ROCK1,phosphorylated myosin phosphatase target-subunit (p-MYPT),phosphorylated MAPK kinase,phosphorylated p38 (p-p38),phosphorylated PKC (p-PKC),and phosphorylated ERM (p-ERM) were determined by western blotting.Results:F-actin reorganization in Ⅳ-infected PMVECs was reversed by XDY-CS treatment,which was accompanied by reduced p-ERM production.The p-ERM protein accumulated at plasma membrane of PMVECs infected with Ⅳ,which was also inhibited by XDY-CS treatment.In addition,XDY-CS treatment drastically reduced the levels of p-p38,ROCK1,p-MYPT,and p-PKC induced by Ⅳ infection in PMVECs.Conclusion:These results show that XDY-CS inhibited influenza-induced F-actin reorganization in PMVECs by down-regulating p-ERM expression via inhibition of the Rho/ROCK,p38 MAPK,and PKC pathways.In conclusion,XDY could reduce the damage to endothelial cytoskeleton induced by Ⅳ infection,thus protecting the barriers of PMVECs.