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目的:分析艳山姜挥发油(EOFAZ)保护脂多糖(LPS)诱导人脐静脉内皮细胞(HUVECs)损伤的主要活性组分。方法:将EOFAZ中含量较高的4个成分(α-蒎烯,β-蒎烯,1,8-桉叶油醇,莰烯)作为考察因素,每个因素选取有或无共2个水平,按L8(27)正交表设计分组与给药。运用四甲基偶氮唑盐(MTT)法和NO检测法,以细胞活力和细胞分泌NO为指标分析EOFAZ的主要药效物质基础。采用单成分分析法测定各成分不同浓度给药保护后细胞活力及培养上清液NO含量,观察4个成分对LPS诱导损伤的HUVECs的保护作用。结果:挥发油中4个成分均能明显提高细胞存活率,保护作用主次顺序为莰烯>α-蒎烯>1,8-桉叶油醇>β-蒎烯,4个成分间均存在明显差异。单成分分析表明4个成分作用于HUVECs后,可提高细胞活力及培养上清液NO含量,且呈一定剂量相关性。结论:EOFAZ对LPS诱导损伤的HUVECs具有保护作用,其中莰烯,α-蒎烯,1,8-桉叶油醇和β-蒎烯为主要药效组分。
OBJECTIVE: To analyze the main active components of Yau Ginger volatile oil (EOFAZ) and lipopolysaccharide (LPS) -induced injury of human umbilical vein endothelial cells (HUVECs). Methods: Four factors (α-pinene, β-pinene, 1,8-eucalyptol, camphene) with high contents in EOFAZ were selected as the study factors. Each factor was selected with or without two levels , According to L8 (27) orthogonal table design and administration. The main pharmacodynamic basis of EOFAZ was analyzed by MTT assay and NO assay using cell viability and NO secretion as indicators. The single factor analysis was used to determine the cell viability and the content of NO in the supernatant after the different concentrations of the components were administered. The protective effects of the four components on LPS-induced HUVECs injury were observed. Results: Four components of volatile oil significantly increased the cell viability. The order of protection was camphene> α-pinene> 1,8-eucalyptol> β-pinene, with significant differences among the four components difference. Single component analysis showed that the four components can increase the cell viability and the content of NO in the supernatant after treated with HUVECs in a dose-dependent manner. CONCLUSION: EOFAZ has a protective effect on HUVECs induced by LPS, among which camphene, α-pinene, 1,8-eucalyptol and β-pinene are the main effective components.