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徐顺洲,陈凌波,阎卉芳,邓常清.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[J].湖南中医药大学学报,2023,43(4):579-584[点击复制] |
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| 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 |
| 徐顺洲,陈凌波,阎卉芳,邓常清 |
| (湖南中医药大学中西医结合学院, 湖南 长沙 410208;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) |
| 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（astragalosideⅠ, ASⅠ）、黄芪甲苷（astragaloside Ⅳ, AS Ⅳ）、毛蕊异黄酮（calycosin, CAL）配伍对血管紧张素Ⅱ（angiotensinⅡ, 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 typeⅠ, COLⅠ）、Ⅲ型胶原（collagen typeⅢ, 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、AS Ⅳ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、AS Ⅳ、CAL敲入后促进MMP2表达的作用增强（P＜0.05或P＜0.01），FMN、AS Ⅳ、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。 |
| 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 |
| DOI：10.3969/j.issn.1674-070X.2023.04.001 |
| 投稿时间：2022-06-13 |
| 基金项目:国家自然科学基金项目（81874406）；湖南省教育厅科学研究项目（18B245）。 |
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| 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 |
| 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) |
| Abstract: |
| 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), astragalosideⅠ (ASⅠ), astragalosideⅣ(ASⅣ) and calycosin (CAL), on the synthesis of extracellular matrix (ECM) by vascular adventitia fibroblasts (VAF) induced by angiotensinⅡ (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 AS I, 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, AS I, and AS Ⅳ, 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, ASⅣ and CAL, the expression of MMP2 was enhanced (P<0.05 or P<0.01), and after the knock-in of FMN, ASⅣ and CAL, the expression of TIMP2 was enhanced (P<0.05 or P<0.01); the compatibility of six active ingredients of Huangqi (Radix Astragali)-Danggui (Radix Angelicae Sinensis) can inhibit the expression of TGF-β1 (P<0.05); after the knock-in of FA, CG and FMN, the expression of TGF-β1 was more inhibited (P<0.05 or P<0.01).Conclusion The compatibility of six active ingredients of Huangqi (Radix Astragali)-Danggui (Radix Angelicae Sinensis) can inhibit the synthesis of ECM by VAF in rats, possibly by regulating TGF-β1, MMP2 and TIMP2. |
| Key words: Huangqi (Radix Astragali) Danggui (Radix Angelicae Sinensis) active ingredients vascular adventitia fibroblasts extracellular matrix |
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