多伞阿魏挥发油化学成分GC-MS分析及D-柠檬烯抗胃癌活性研究

GC-MS Analysis of Volatile Oil from Ferula Ferulaeoides and Anti-gastric Cancer Activity of D-Limonene in Vitro

关键词:  多伞阿魏  挥发油  GC-MS  D-柠檬烯  抗胃癌

Key words:  Ferula ferulaeoides  volatile oil  GC-MS  D-limonene  anti-gastric cancer

摘要:

目的 分析多伞阿魏新鲜药材挥发油的化学成分，初步探讨D-柠檬烯体外抗胃癌作用。方法 采用GC-MS分析多伞阿魏挥发油的化学成分；采用MTT法检测D-柠檬烯对人胃癌细胞AGS和SGC-7901的增殖抑制作用；采用Hoechst33258荧光染色法观察D-柠檬烯对人胃癌细胞AGS的影响；应用流式细胞仪检测D-柠檬烯对人胃癌细胞AGS凋亡的影响。结果 多伞阿魏挥发油主要的化学成分有α-蒎烯（6.76%）、崁烯（8.38%）、α-月桂烯（4.66%）、3-蒈烯（4.84%）、D-柠檬烯（52.84%）、乙酸龙脑酯（5.24%）等。MTT法检测结果显示，与空白对照组比较，D-柠檬烯对人胃癌细胞AGS和SGC-7901均具有不同程度的增殖抑制作用，呈现剂量依赖关系，其中D-柠檬烯对人胃癌细胞AGS的增殖抑制作用较强，IC₅₀为（0.87±0.05）mmol·L^-1；D-柠檬烯作用于人胃癌细胞AGS后，细胞核被Hoechst 33258染色呈亮蓝色，随着药物浓度的增加蓝色荧光越强；流式细胞凋亡检测结果显示，与空白对照组比较，D-柠檬烯可诱导人胃癌细胞AGS发生凋亡，呈现剂量依赖关系。结论 多伞阿魏挥发油的主要物质基础为单萜类、倍半萜类化合物，其中D-柠檬烯的相对百分含量最高；D-柠檬烯对人胃癌细胞AGS和SGC-7901均具有一定的增殖抑制作用，其中D-柠檬烯对人胃癌细胞AGS的诱导凋亡活性最强。

Abstract:

OBJECTIVE To analyze the chemical constituents of the volatile oil from the fresh herbal medicine of Ferula ferulaeoides, and explore the anti-gastric cancer activity of D-limonene in vitro. METHODS GC-MS was used to analyze the chemical constituents of volatile oil of Ferula ferulaeoides; the MTT method was used to detect the proliferation inhibition effect of D-limonene on human gastric cancer cells AGS and SGC-7901; Hoechst 33258 fluorescent staining was used to observe the effect of D-limonene on the human gastric cancer cells AGS; the apoptosis of human gastric cancer cells AGS induced by D-limonene was detected by flow cytometry. RESULTS The results showed that α-pinene(6.76%), camphene(8.38%), α-myrcene(4.66%), 3-carene(4.84%), D-limonene(52.84%), bornyl acetate(5.24%), ect, were the main chemical constituents in the volatile oil of Ferula ferulaeoides. The MTT assay results showed that D-limonene had different degrees of proliferation inhibition effects on human gastric cancer cells AGS and SGC-7901, and showed a dose-dependent relationship. D-limonene had a strong proliferation inhibition effect on the human gastric cancer cell AGS, and its IC₅₀ was (0.87±0.05)mmol·L^-1. The cell nucleus of human gastric cancer cell lines AGS was stained by Hoechst 33258, and the blue fluorescence was stronger with the increase of drug concentration. The results of flow cytometry showed that D-limonene induced apoptosis of human gastric cancer cells AGS, and showed a dose-dependent relationship. CONCLUSION Monoterpenoids and sesquiterpenoids are main chemical constituents of the volatile oil of Ferula ferulaeoides. D-limonene is account for highest percentage of the total constituents. D-limonene has proliferation inhibition effect on human gastric cancer cells AGS and SGC-7901, the cytotoxicity activity of D-limonene on human gastric cancer cell AGS is strongest.