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)reorganiza...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),p38 MAPK,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.展开更多
Objective:To study the protective effect of the Mixture of Shengmai Powder and Danshen Decoction(生脉散丹参饮合剂,abbreviated as the Mixture) in the rat model with type 2 diabetic cardiomyopathy (DCM).Methods:Fo...Objective:To study the protective effect of the Mixture of Shengmai Powder and Danshen Decoction(生脉散丹参饮合剂,abbreviated as the Mixture) in the rat model with type 2 diabetic cardiomyopathy (DCM).Methods:Forty-two SD rats with DCM model,established by the combination of insulin resistance by a high-fat diet with the damage of pancreatic islet p cells by intraperitoneal injection of high dose streptozotocin (50 mg/kg) once,were evaluated in the damage of the myocardium by electrocardiogram at the end of 12 weeks of grouping and intervention administration;the extent of damage in the myocardial subcellular structure was observed by electron microscopy;the content of myocardial collagen in the left cardiac ventricle was quantified by Masson staining test;the myocardial cell apoptosis was determined by TUNEL;the changes in the mRNA expression levels of thrombospodin-1(TSP-1) and tribbles homolog 3(TRB-3) by real-time quantitative PCR,the expression levels of myocardial TSP-1,tumor growth factorβ1(TGF-β1),TRB-3,and chymase were detected by immunohistochemistry,and the changes in the expression levels of myocardial TSP-1,active-TGF-β1 (A-TGF-β1) and latent-TGF-β1(L-TGF-β1) protein were tested by Western blotting.Results:Compared with the control group,the myocardial tissue was less damaged,and the extent of damage in the myocardial subcellular structure was less;the collagen fiber content and the cell apoptosis were reduced;the expression levels of TSP-1 mRNA and TRB-3 mRNA,the expression levels of myocardial TSP-1,TGF-β1,TRB-3,and chymase,as well as the average expression levels of the myocardial TSP-1,A-TGFβ1,and L-TGF-β1 protein were decreased in the Mixture group.Conclusion:The Mixture of Shengmai Powder and Danshen Decoction could inhibit the process of myocardial fibrosis in the rat myocardium of DCM through multiple pathways and significantly delay the genesis and progress of DCM in hyperglycemic rats.展开更多
基金This work was supported by funding from the National Natural Science Foundation of China(Grant Nos.81473520 and 81102697).
文摘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),p38 MAPK,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.
基金Supported by the First Grade of China Postdoctoral Science Foundation(No.20070410129)the Special Fund by China Postdoctoral Science Foundation(No.200801166)+1 种基金the Major Project of Beijing Municipal Science and Technology Committee (No.H020920010330)the Subject of Science and Technology Plan of Beijng Municipal Science and Technology Committee(No.D08050703020802)
文摘Objective:To study the protective effect of the Mixture of Shengmai Powder and Danshen Decoction(生脉散丹参饮合剂,abbreviated as the Mixture) in the rat model with type 2 diabetic cardiomyopathy (DCM).Methods:Forty-two SD rats with DCM model,established by the combination of insulin resistance by a high-fat diet with the damage of pancreatic islet p cells by intraperitoneal injection of high dose streptozotocin (50 mg/kg) once,were evaluated in the damage of the myocardium by electrocardiogram at the end of 12 weeks of grouping and intervention administration;the extent of damage in the myocardial subcellular structure was observed by electron microscopy;the content of myocardial collagen in the left cardiac ventricle was quantified by Masson staining test;the myocardial cell apoptosis was determined by TUNEL;the changes in the mRNA expression levels of thrombospodin-1(TSP-1) and tribbles homolog 3(TRB-3) by real-time quantitative PCR,the expression levels of myocardial TSP-1,tumor growth factorβ1(TGF-β1),TRB-3,and chymase were detected by immunohistochemistry,and the changes in the expression levels of myocardial TSP-1,active-TGF-β1 (A-TGF-β1) and latent-TGF-β1(L-TGF-β1) protein were tested by Western blotting.Results:Compared with the control group,the myocardial tissue was less damaged,and the extent of damage in the myocardial subcellular structure was less;the collagen fiber content and the cell apoptosis were reduced;the expression levels of TSP-1 mRNA and TRB-3 mRNA,the expression levels of myocardial TSP-1,TGF-β1,TRB-3,and chymase,as well as the average expression levels of the myocardial TSP-1,A-TGFβ1,and L-TGF-β1 protein were decreased in the Mixture group.Conclusion:The Mixture of Shengmai Powder and Danshen Decoction could inhibit the process of myocardial fibrosis in the rat myocardium of DCM through multiple pathways and significantly delay the genesis and progress of DCM in hyperglycemic rats.