黄芪-当归6种活性成分配伍对大鼠血管外膜成纤维细胞合成细胞外基质的影响  

Effects of compatibility of 6 active ingredients of Huangqi(Radix Astragali)-Danggui(Radix Angelicae Sinensis)on extracellular matrix synthesis by vascular adventitia fibroblasts in rats

作　　者：徐顺洲 陈凌波[1] 阎卉芳 邓常清[1] XU Shunzhou;CHEN Lingbo;YAN Huifang;DENG Changqing(School of Integrated Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha,Hunan 410208,China;Department of Cardiology,The First Hospital of Hunan College of Chinese Medicine,Zhuzhou,Hunan 412000,China)

机构地区：[1]湖南中医药大学中西医结合学院,湖南长沙410208 [2]湖南中医药高等专科学校附属第一医院心内科,湖南株洲412000

出　　处：《湖南中医药大学学报》2023年第4期579-584,共6页Journal of Hunan University of Chinese Medicine

基　　金：国家自然科学基金项目(81874406);湖南省教育厅科学研究项目(18B245)。

摘　　要：目的观察黄芪-当归活性成分阿魏酸(ferulic acid,FA)、毛蕊异黄酮苷(calycosinglucoside,CG)、芒柄花素(formononetin,FMN)、黄芪皂苷Ⅰ(astragaloside Ⅰ,ASⅠ)、黄芪甲苷(astragaloside Ⅳ,ASⅣ)、毛蕊异黄酮(calycosin,CAL)配伍对血管紧张素Ⅱ(angiotensin Ⅱ,AngⅡ)诱导的大鼠血管外膜成纤维细胞(vascular adventitia fibroblasts,VAF)合成细胞外基质(extracellular matrix,ECM)的影响。方法以AngⅡ诱导VAF增殖模型,采用目标成分“敲除/敲入”的方法,将细胞分为空白组、模型组、IC10配伍组、某一活性成分敲除组、某一活性成分敲入组,分别检测细胞及培养液中纤维连接蛋白(fibronectin,FN)、层粘连蛋白(laminin,LN)、Ⅰ型胶原(collagen type Ⅰ,COLⅠ)、Ⅲ型胶原(collagen type Ⅲ,COLⅢ)含量,并检测细胞基质金属蛋白酶2(matrix metalloproteinase 2,MMP2)、基质金属蛋白酶组织抑制剂2(tissue inhibitor of metalloproteinase 2,TIMP2)、转化生长因子β1(transforming growth factor-β1,TGF-β1)蛋白表达,研究黄芪-当归6种活性成分配伍对VAF合成ECM的影响。结果黄芪-当归6种活性成分配伍抑制VAF合成FN、LN、COLⅠ、COLⅢ(P<0.01)。FA、CG、FMN、ASⅠ敲入后抑制FN、LN合成的作用增强(P<0.05或P<0.01),FA、CAL、FMN、ASⅠ、ASⅣ敲入后抑制COLⅠ、COLⅢ合成的作用增强(P<0.05或P<0.01)。黄芪-当归6种活性成分配伍促进MMP2、TIMP2的表达(P<0.01),FA、CG、FMN、ASⅣ、CAL敲入后促进MMP2表达的作用增强(P<0.05或P<0.01),FMN、ASⅣ、CAL敲入后促进TIMP2表达的作用增强(P<0.05或P<0.01);黄芪-当归6种活性成分配伍抑制TGF-β1表达(P<0.05),FA、CG、FMN敲入后抑制TGF-β1表达的作用增强(P<0.05或P<0.01)。结论黄芪-当归6种活性成分配伍可抑制VAF合成ECM,其作用可能是通过调节TGF-β1、MMP2、TIMP2发挥的。Objective To observe the effects of 6 active components of Huangqi(Radix Astragali)-Danggui(Radix Angelicae Sinensis),namely ferulic acid(FA),calycosinglucoside(CG),formononetin(FMN),astragaloside Ⅰ(ASⅠ),astragaloside Ⅳ(ASⅣ)and calycosin(CAL),on the synthesis of extracellular matrix(ECM)by vascular adventitia fibroblasts(VAF)induced by angiotensin Ⅱ(AngⅡ)in rats.Methods By employing VAF proliferation model induced by AngⅡ,and the target component"knock-out/knock-in"method,the cells were divided into blank group,model group,10% inhibitory concentration compatibility group,active component knock-out group,and active component knock-in group.Then,the content of fibronectin(FN),laminin(LN),collagen typeⅠ(COLⅠ),and collagen type Ⅲ(COLⅢ)in cells and culture medium were measured respectively.The expression levels of matrix metalloproteinase2(MMP2),tissue inhibitor of matrix metalloproteinase2(TIMP2),and transforming growth factor-β1(TGF-β1)were detected.Based on the above,we studied the effects of compatibility of six active components of Huangqi(Radix Astragali)-Danggui(Radix Angelicae Sinensis)on extracellular matrix synthesis by vascular adventitia fibroblasts in rats.Results The compatibility of 6 active ingredients of Huangqi(Radix Astragali)-Danggui(Radix Angelicae Sinensis)can inhibit the synthesis of FN,LN,COLⅠ and COLⅢ by VAF(P<0.01);after the knock-in of FA,CG,FMN and ASⅠ,the synthesis of FN and LN was more inhibited(P<0.05 or P<0.01);and after the knock-in of FA,CAL,FMN,ASⅠ,and ASⅣ,the synthesis of COLⅠ and COLⅢ was more inhibited(P<0.05 or P<0.01).The compatibility of 6 active ingredients of Huangqi(Radix Astragali)-Danggui(Radix Angelicae Sinensis)can promote the expression of MMP2 and TIMP2(P<0.01);after the knock-in of FA,CG,FMN,ASⅣand CAL,the expression of MMP2 was enhanced(P<0.05 or P<0.01),and after the knock-in of FMN,ASⅣand CAL,the expression of TIMP2 was enhanced(P<0.05 or P<0.01);the compatibility of six active ingredients of Huangqi(Radix Astragali)-Danggu

关 键 词：黄芪 当归 活性成分 血管外膜成纤维细胞 细胞外基质 

分 类 号：R285.5[医药卫生—中药学]