目的 评价香附四物汤及其不同分离部位对大鼠卵巢颗粒细胞增殖的影响,并对活性部位成分进行分析.方法 体外培养大鼠卵巢颗粒细胞,将香附四物汤的不同分离方法 所得提取物以2、20、200μg/mL 3个终浓度分别加入培养体系中,通过四甲基偶氮唑盐微量酶反应比色法(MTT法)检测细胞增殖情况,观察其对卵巢颗粒细胞生长的影响.采用超高效液相-四级杆串联飞行时间-质谱(UPLC-QTOF-MS)分析鉴定活性部位的成分.结果 香附四物汤挥发油、水煎液、醇沉沉淀、醇沉上清液及醇沉分子量大于5万的部位对卵巢颗粒细胞均具有明显的促进增殖作用,并呈现剂量依赖性.其中水提物大孔树脂醇洗脱部位在200μg/mL剂量下30%及40%部位表现出显著的促进增殖作用.从醇沉上清液中分析鉴定了16种化合物.结论 香附四物汤对离体培养的卵巢颗粒细胞增殖具有明显的促进作用,其调节机体性激素生成作用可能与促进卵巢颗粒细胞生长效应相关;活性部位醇沉上清液的化学组分主要为萜类、有机酸类和生物碱类成分.%OBJECTIVE To investigate the effects of Xiangfu Siwu Decoction and its separated parts on the proliferation of ovary granulosa cells of rats, and to analyze the chemical components of the bioactive fraction. METHODS Xiangfu Siwu Decoction as well as its extractives obtained through different separation methods at the concentration of 2,20,200μg/mL was added into the rat ovary granulosa cells cultivated in vitro to measure the proliferation of the cells by MTT.Bioactive components were analyzed by UPLC-QTOF-MS. RESULTS Volatile oil, water extraction, ethanol deposition,ethanol supernatant and fraction with molecular weight higher than 50 000 Da significantly improved the proliferation of granulosa cells in a dose-dependent manner. 30 ％ and 40％ ethanol eluted fraction also showed remarkable promotion on proliferation at the dose of 200 μg/mL. Altogether 16 compounds in ethanol supernatant were identified. CONCLUSION Xiangfu Siwu Decoction as well as its extractives encourages the proliferation of rat ovary granulosa cells, whose mechanism may lie in its regulation of steroidogenesis. Terpenoids, organic acids and alkaloids are identified in the effective fraction ethanol supernatant.
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