Liuwei Dihuang soft capsules inhibits phenotypic conversion of VSMCs via up-regulating expression of myocardin by promoting interaction between ERα and SRC3  

作　　者：MENG Qing-hai YU Xi-chao CHEN Qi KONG Xue-yun CHENG Peng LI Yu BIAN Hui-min 

机构地区：[1]Nanjing University of Chinese Medicine

出　　处：《中国药理学与毒理学杂志》2019年第9期694-695,共2页Chinese Journal of Pharmacology and Toxicology

基　　金：National Natural Science Foundation of China(81773190;81774029);Jiangsu Provincial Science and Technology Department Social Development Fund(BE2011846)

摘　　要：OBJECTIVE To reveal the under mechanism of how Liuwei Dihuang(LWDH)inhibits the phenotypic con⁃version of VSMCs.METHODS 24 ApoE-/-mice were divided into 4 groups:sham group,model group,E2 group,and LWDH group.Six C57BN/L6 mice were used as control group.The primary VSMCs were divided into control group,model group,E2 group,LWDH group,LWDH+MPP group,and LWDH+PHTPP group with or without control siRNA,ERαsiRNA,ERβsiRNA,and myocardin siRNA.Oil red staining was used to evaluate the lipid deposition in the cardiac aorta.Serum chemistry analysis to test serum TG,TC,LDL,and HDL.Immunofluorescence staining was used to testα-SMA,osteopontin and F-actin.Immunohistochemical staining was performed to check out the myocardin in the cardiac aorta.The mRNA levels ofα-SMA,osteopontin,ERα,ERβ,SRC3 and myocardin were detected by real time-PCR,and the protein expression levels of them were detected by Western blotting.Co-immunoprecipitation was proceeded to test the interaction between ERαand SRC3 and SRC3 and myocardin.Flow cytometry was used to check out the cell cycle.Wound healing assay and Transwell were managed to evaluate the migration capacity of VSMCs.RESULTS In vivo administration of LWDH suppressed atherosclerotic symptoms,decreases phenotypic marker of vascular endothelial cell,and increases phenotypic marker of VSMC in ovariectomized ApoE-/-female mice.Moreover,LWDH significantly increased the mRNA and protein expression levels of ERα,ERβ,SRC3 and myocardin in the cardiac aorta of ovariecto⁃mized ApoE-/-female mice.In vitro,LWDH altered cell cycle and reduced the elevated cyclinD protein expression migra⁃tion capacity and in the model VSMCs.In addition,LWDH inhibited phenotypic conversion and promoted the expression of ER,SRC3,and myocardin of the primary VSMC phenotypic conversion model.Inhibition of ERαalmost completely eliminated the impacts of LWDH onα-SMA and osteopontin.Furthermore,LWDH promoted the interaction between ERαand SRC3 and up-regulated the co-activation of SRC3 and myocardin.CONCLUSIOOBJECTIVE To reveal the under mechanism of how Liuwei Dihuang(LWDH) inhibits the phenotypic conversion of VSMCs. METHODS 24 Apo E-/-mice were divided into 4 groups: sham group, model group, E2 group, and LWDH group. Six C57 BN/L6 mice were used as control group. The primary VSMCs were divided into control group,model group, E2 group, LWDH group, LWDH+MPP group, and LWDH+PHTPP group with or without control si RNA,ERα si RNA, ERβ si RNA, and myocardin si RNA. Oil red staining was used to evaluate the lipid deposition in the cardiac aorta. Serum chemistry analysis to test serum TG, TC, LDL, and HDL. Immunofluorescence staining was used to testα-SMA, osteopontin and F-actin. Immunohistochemical staining was performed to check out the myocardin in the cardiac aorta. The mRNA levels of α-SMA, osteopontin, ERα, ERβ, SRC3 and myocardin were detected by real time-PCR, and the protein expression levels of them were detected by Western blotting. Co-immunoprecipitation was proceeded to test the interaction between ERα and SRC3 and SRC3 and myocardin. Flow cytometry was used to check out the cell cycle.Wound healing assay and Transwell were managed to evaluate the migration capacity of VSMCs. RESULTS In vivo administration of LWDH suppressed atherosclerotic symptoms, decreases phenotypic marker of vascular endothelial cell, and increases phenotypic marker of VSMC in ovariectomized Apo E-/-female mice. Moreover, LWDH significantly increased the mRNA and protein expression levels of ERα, ERβ, SRC3 and myocardin in the cardiac aorta of ovariectomized Apo E-/-female mice. In vitro, LWDH altered cell cycle and reduced the elevated cyclin D protein expression migration capacity and in the model VSMCs. In addition, LWDH inhibited phenotypic conversion and promoted the expression of ER, SRC3, and myocardin of the primary VSMC phenotypic conversion model. Inhibition of ERα almost completely eliminated the impacts of LWDH on α-SMA and osteopontin. Furthermore, LWDH promoted the interaction between ERα and SRC3 and up-regula

关 键 词：Liuwei Dihuang VSMC atherosclerosis MENOPAUSE ER MYOCARDIN 

分 类 号：R73[医药卫生—肿瘤]