补阳还五汤苷类有效组分及其有效成分配伍对血管平滑肌细胞增殖及相关信号转导通路的影响  

Effects of glycosides components and combinations of Buyang Huanwu decoction on vascular smooth muscle cells proliferation and related signaling pathway

作　　者：曹浪[1] 邓常清[1] 

机构地区：[1]湖南中医药大学分子病理实验室,中西医结合心脑疾病防治湖南省重点实验室,细胞生物学与分子技术湖南省高校重点实验室,湖南长沙410208

出　　处：《中国中药杂志》2016年第10期1889-1897,共9页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81072750);湖南省高校科技创新团队项目;湖南省自然科学析群体项目

摘　　要：研究补阳还五汤苷类有效组分和其主要有效成分黄芪甲苷、苦杏仁苷、芍药苷及其配伍抗血管平滑肌细胞(vascular smooth muscle cell,VSMC)增殖的作用,阐明其抗VSMC增殖的主要药效物质,并从细胞信号转导途径研究其作用机制。大鼠口服给药后制备含药血浆。培养大鼠主动脉VSMC,采用血小板衍生生长因子(platelet derived growth factor,PDGF)刺激VSMC增殖,加入含药血浆,测定细胞增殖活性、细胞周期及其相关蛋白表达、细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)、磷脂酰肌醇3激酶(phos-phatidylinositol-3-kinase/protein kinase B,PI3K/Akt)、Janus激酶/信号转导与激活因子(Janus kinase/signal transducer and activator of transcription,JAK/STAT)信号通路相关蛋白表达。结果显示,PDGF刺激后,VSMC增殖活性增强(P<0.01),G0/G1期细胞减少(P<0.01),S/M期细胞增多(P<0.01),同时细胞周期相关蛋白PCNA,cyclin D1蛋白表达增强(P<0.01)。苷类组分、3个主要有效成分黄芪甲苷、芍药苷、苦杏仁苷及其配伍均可抑制细胞增殖(P<0.05或P<0.01),作用呈量效关系和时效关系;且可使G0/G1期细胞增多(P<0.01),S/M期细胞减少(P<0.01),PCNA,cyclin D1蛋白表达下调(P<0.01),3个有效成分配伍的作用强于单个有效成分(P<0.05)。PDGF诱导VSMC增殖后,p-ERK1/2表达增强(P<0.01),PI3K表达降低(P<0.01),p-PI3K表达增强(P<0.01),STAT3表达降低(P<0.01),p-STAT3表达增高(P<0.01)。苷类有效组分和主要有效成分黄芪甲苷、芍药苷、苦杏仁苷及其配伍可使p-ERK1/2表达降低(P<0.05),PI3K表达升高(P<0.01),p-PI3K表达降低(P<0.05或P<0.01),STAT3表达升高(P<0.01),p-STAT3表达下调(P<0.05或P<0.01)。上述结果表明PDGF可诱导VSMC细胞周期转化,使VSMC增殖。其机制与ERK,PI3K/Akt,JAK/STAT信号通路激活有关。补阳还五汤苷类有效组分和其主要有效成分黄芪甲苷、芍药苷、苦杏仁苷及其配伍可抑制VSMC细胞周期转换,具有抗VSMC�This paper was aimed to explore the effects of glycosides,the effective component of Buyang Huanwu decoction,and its main active components such as astragaloside Ⅳ,amygdalin,peoniflorin and their combinations on vascular smooth muscle cells（ VSMC） proliferation,clarify the major active materials of anti-VSMC proliferation and investigate the mechanisms via the signal transduction pathway. Plasma containing drug was prepared via oral administration in rats. VSMCs of rats aorta were cultured,and then VSMC proliferation was stimulated by using platelet derived growth factor（ PDGF）. The plasma containing drug was added to detect the activity of cell proliferation,cell cycle and related protein expressions of signaling pathway such as extracellular signal-regulated kinase（ ERK）,phos-phatidylinositol-3-kinase/protein kinase B（ PI3K/Akt） and Janus kinase/signal transducer and activator of transcription（ JAK/STAT）. After being stimulated by PDGF,the proliferation activity of VSMC was strengthened（ P〈0. 01）,G0/ G1 phase cells were decreased（ P〈0. 01）,S / M phase cells were increased（ P〈0. 01）,and Pc NA,cyclin D1 protein expressions related to cell cycle were up-regulated（ P〈0. 01）. Glycosides,astragaloside Ⅳ,amygdalin,peoniflorin and their combinations could inhibit the cell proliferation（ P〈0. 05 or P〈0. 01） in a dose-effect relationship and time-effect relationship. They could increase G0/ G1 phase cells（ P〈0. 01）,decrease S/M phase cells（ P〈0. 01）,and down-regulate the protein expressions of PCNA,cyclin D1（ P〈0. 01）; and the effects of the combinations were greater than those of single active component（ P〈0. 05）. After VSMC proliferation was induced by PDGF,p-ERK1 /2 expression was increased（ P〈0. 01）,PI3 K expression was down-regulated while p-PI3 K expression was up-regulated（ all P〈0. 01）,and STAT3 expression was reduced while p-STAT3 expression was increased（ all P〈0. 01）. Glycosides,astragaloside Ⅳ,amygdalin,peonifl

关 键 词：补阳还五汤 苷类有效组分 黄芪甲苷 苦杏仁苷 芍药苷 血管平滑肌细胞增殖 细胞周期 

分 类 号：R285.5[医药卫生—中药学]