目的 研究齐墩果酸（OA）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，并探讨其可能的作用机制。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（10-6 mol·L-1）组、Ang II（10-6 mol·L-1）+OA（60，80，100 μmol·L-1）组。细胞长至融合状态时，先加入OA（60，80， 100 μmol·L-1）预处理1 h，然后加入Ang II（10-6 mol·L-1）作用24 h。应用二甲基四唑氮蓝（MTT）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型胶原含量，蛋白质印迹法（Western blotting）和激光扫描共聚焦显微镜分别检测各组CFs中结缔组织生长因子（connective tissue growth factor，CTGF）蛋白表达和活性氧（ROS）变化。结果 Ang II能诱导CFs增殖，并能增加细胞上清中I型胶原的含量，同时CTGF的蛋白表达及CFs中ROS水平也显著增加。OA在60~100 μmol·L-1浓度范围内呈浓度依赖性抑制Ang II诱导的CFs增殖，并且OA能减弱Ang II诱导的CTGF和ROS表达。结论 OA可通过ROS-CTGF途径抑制Ang II诱导的CFs增殖和胶原生成。

Objective To investigate the inhibitive effect of oleanolic acid（OA） on cardiac fibroblasts（CFs） proliferation induced by angiotension II（Ang II） and to explore the related mechanism. Methods Rat CFs were isolated by the methods of digestive enzyme and differential adhesion. When the rat CFs became cell fusion，OA （60，80，100 μmol·L-1） was added into the medium. One hour later，Ang II（10-6 mol·L-1） was added into the medium for co-culture for 24 hours. Cell proliferation was measured by MTT assay. The content of type I collagen in the culture medium was measured by enzyme-linked immunosorbent assay（ELISA）. Western blotting method was used to detect the protein level of connective tissue growth factor（CTGF），and cellular reactive oxygen species（ROS） levels were detected by confocal laser scanning microscope. Results Ang II significantly induced CFs proliferation，increased the content of type I collagen，elevated CTGF protein expression and ROS levels in CFs. OA at the dose of 60~100 μmol·L-1 showed inhibitory effect on CFs proliferation induced by Ang II in dose-dependent manner. OA also inhibited the expression levels of CTGF and ROS induced by Ang II. Conclusion OA can inhibit the proliferation of CFs induced by Ang II and the formation of collagen through ROS-CTGF pathway.

R285.5

国家自然科学基金（31300946）；泸州医学院附属医院人才基金（12290）；泸州市科技计划项目（12330）。