目的 研究桃叶珊瑚苷(AU)对血管紧张素Ⅱ(Ang Ⅱ)诱导的大鼠心肌成纤维细胞(CFs)增殖的影响及作用机制.方法 采用胰酶消化法及差速贴壁法培养大鼠CFs,实验分为空白对照组、Ang Ⅱ(1×10-6 mol/L)干预组、低浓度AU 干预组[Ang Ⅱ(1×10-6 mol/L)+AU(1 μmol/L)]、中浓度AU 干预组[Ang Ⅱ(1×10-6 mol/L)+AU(10 μmol/L)]、高浓度AU 干预组[Ang Ⅱ(1×10-6 mol/L)+AU(100 μmol/L)].用CCK8 法测定细胞增殖活性,酶联免疫吸附法(ELISA)测定细胞上清中Ⅰ、Ⅲ型胶原含量,荧光酶标仪检测活性氧(ROS)水平,Western blot 法检测p38MAPK和p-p38MAPK蛋白表达.结果 与Ang Ⅱ干预组比较,中、高浓度AU 干预组CFs 增殖和p38MAPK 磷酸化被明显抑制(P ＜ 0.05).与Ang Ⅱ干预组比较,低、中、高浓度AU 干预组ROS、Ⅰ、Ⅲ 型胶原的生成明显减少(P ＜ 0.05).结论 AU 可通过ROS-MAPK 通路途径抑制Ang Ⅱ诱导的CFs 增殖和胶原生成.%Objective To investigate the effect and mechanism of aucubin (AU) on cardiac fibroblasts (CFs) proliferation induced by angiotension Ⅱ (Ang Ⅱ). Methods Rat CFs were cultured by the methods of trypsin digestion and differential adhesion. They were divided into blank control group, Ang Ⅱ (1×10-6 mol/L) intervention group, low concentration of AU intervention group [Ang Ⅱ (1×10-6 mol/L) + AU (1 μmol/L)], medium concentration of AU intervention group [Ang Ⅱ (1×10-6 mol/L) + AU (10 μmol/L)], high concentration of AU intervention group [Ang Ⅱ (1×10-6 mol/L) + AU (100 μmol/L)]. CCK8 method was used to test cell proliferation activity, collagen type Ⅰ, Ⅲ content in cell supernatant were determed by enzyme linked immunosorbent (ELISA), fluorescence enzyme standard instrument was used to detect reactive oxygen species (ROS) level, Western blot method was used to detect p38MAPK and p-p38MAPK protein expression. Results Compared with Ang Ⅱ intervention group, CFs proliferation and p38MAPK phosphorylation were significantly inhibited in the medium and high concentration AU intervention groups (P ＜ 0.05). Compared with Ang Ⅱ intervention group, low, medium and high concentration of AU intervention group ROS, the production of collagen type Ⅰ, Ⅲ decreased significantly (P ＜ 0.05). Conclusion AU can restrain by ROS-MAPK pathway Ang Ⅱ induction of CFs proliferation and collagen formation.
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