目的 探讨温阳活血方对自噬损伤内皮细胞保护作用的物质基础以及对凋亡相关因子Activate Caspase-3、自噬相关蛋白泛素结合蛋白P62以及分化抑制因子1（ID1）表达的影响。方法 将温阳活血方不同极性提取物（石油醚提取物、乙酸乙酯提取物、乙醇提取物和水提取物）作用于过氧化氢诱导的自噬性损伤内皮细胞模型，应用二苯基四氮唑溴盐（MTT）法观察内皮细胞的活性，用气相色谱-质谱联用技术（GS-MS）分析有效部位的化学成分，Western blot法检测有效部位对Activate Caspase-3、P62、ID1表达的影响。结果 温阳活血方石油醚部位可提高内皮细胞活性；GS-MS检测发现石油醚部位主要成分有4-（4-羟基-3-甲氧基苯基）-2-丁酮、3-丁烯基苯酞、Z-藁本内酯、E-藁本内酯、2，10-冰片二醇、Z，Z-9，12-亚油酸、6-姜酚、8-姜酚等；Western blot检测发现，石油醚部位能上调ID1、P62的表达，抑制Activate Caspase-3的表达。结论 石油醚部位是温阳活血方对抗内皮细胞自噬性死亡的活性部位，其通过降低内皮细胞内Activate Caspase-3的表达，提高P62和ID1的表达，以对抗内皮细胞的自噬性死亡。

Objective To investigate the active fraction of Wenyang Huoxue Decoction in protecting human umbilical vein endothelial cells（HUVECs） from autophagic cell deaths induced by hydrogen peroxide，and to research its effect on the expression of activated caspase-3，ubiquitin-binding protein P62 and inhibitor of differentiation 1（ID1）. Methods Wenyang Huoxue Decoction was boiled in Soxhlet extractor，and was extracted separately with the solvents of petroleum ether（F1），acetic ether（F2），ethanol（F3） and water（F4）. HUVECs with autophagic cell deaths induced by hydrogen peroxide were cocultured with the different extraction fractions of petroleum ether fraction，acetic ether fraction，ethanol fraction and water fraction. And then methyl thiazolyl tetrazolium assay was used to observe the activities of HUVECs，and the components of active fraction were analyzed by gas chromatography-mass spectrogram（GC-MS）. Western blotting was recruited to analyze the effect of active fraction on the expression of activated caspase-3，P62 and ID1. Results In our study，petroleum ether fraction（F1） was the active fraction of Wenyang Huoxue decoction to protect HUVECs from autophagic deaths. The results of GC-MS showed that main ingredients of F1 were 4-（4-hydroxy-3-methoxyphenyl）-2-butanone，3-butylidenephthalide，Z-ligustilide，E-ligustilide，2，10 -bornyl glycol，Z，Z-9，12-octadecadienoic acid，6-gingerol，8- gingerol. Western blotting analysis showed that F1 could increase the expression of ID1 and P62，and inhibit the expression of activated caspase-3 in HUVECs. Conclusion Petroleum ether fraction（F1） is the active fraction of Wenyang Huoxue Decoction to protect HUVECs from autophagic cell deaths，and its mechanism is related to the decrease of expression of activated caspase-3 in HUVECs，and the increase of the expression of P62and ID1.

R284.1

广西壮族自治区自然科学基金重点项目（2010GXNSFD013051）。