目的 从尖叶假龙胆不同极性部位中寻找对H₂O₂诱导PC12细胞氧化损伤具有较强保护作用的化合物。方法 采用活性追踪分离的方式，通过H₂O₂诱导PC12细胞损伤建立氧化应激损伤模型，利用实时细胞分析技术（real-time cell analysis，RTCA）研究尖叶假龙胆不同极性部位对H₂O₂诱导PC12细胞氧化损伤的保护作用；采用HPLC法测定具有活性的单体化合物的质量分数，并确定其化学结构。结果 尖叶假龙胆正丁醇和醋酸乙酯部位对H₂O₂诱导PC12细胞氧化损伤具有保护作用。同时，从尖叶假龙胆正丁醇和醋酸乙酯部位中分离得到的化合物1、2、3、5也对H₂O₂诱导PC12细胞氧化损伤具有保护作用。其中化合物1和2的保护作用在3.125～50.000 μmol/L呈一定剂量依赖性，浓度为50.000 μmol/L时，对H₂O₂诱导PC12细胞氧化损伤的保护作用最强；而化合物3和5的保护作用在3.125～50.000 μmol/L内不具有剂量依赖性，浓度分别为25.000、3.125 μmol/L时，对H₂O₂诱导PC12细胞氧化损伤的保护作用最强。经结构鉴定，化合物1、2、3、5分别为雏菊叶龙胆酮、去甲基雏菊叶龙胆酮、当药醇苷及去甲基当药醇苷。结论 化合物1、2、3、5对H₂O₂诱导PC12细胞氧化损伤具有保护作用，可能是尖叶假龙胆抗氧化的主要活性成分。

Objective To investigate the compounds of Gentianella acuta with strong protective effect on oxidative damage in PC12 cells induced by H₂O₂. Methods Damage model of PC12 cells was established by H₂O₂ and real-time cell analysis (RTCA) was utilized to evaluate the protective effect of extracts and compounds. Meanwhile, bioassay-guided method was applied to separating effective components, and HPLC was used for the determination and structure confirmation of compounds. Results Fractions of n-butanol and acetic ether showed protective effect on oxidative damage in PC12 cells induced by H₂O₂. The anti-oxidant activity of compounds 1 and 2 showed dose-dependent manner in the scope of 3.125—50.000 μmol/L, and the protective effect was the strongest at the concentration of 50.000 μmol/L. Compounds 3 and 5 showed the best protective effect at the concentration of 25.000 μmol/L and 3.125 μmol/L respectively. Compounds 1, 2, 3, and 5 were identified as bellidifolin, demethylbellidifolin, swertianolin and norswertianolin, respectively. Conclusion Compounds 1, 2, 3, and 5 show protective effect on oxidative damage in PC12 cells induced by H₂O₂, maybe the main active components in G. acuta. But the mechanism is still uncertain, and further exploration is imperative.

国家自然科学基金资助项目（81160504，81303306）；天津市应用基础与前沿技术研究计划（15JCQNJC13400）