化州柚花挥发油的GC-MS分析及其对肺癌细胞A549增殖、迁移的影响研究

GC-MS Analysis of Volatile Oil from the Flowers of Citrus grandis and Study on Effects of It on the Proliferation and Migration of Lung Cancer A549 Cells

目的:提取化州柚花挥发油,鉴定其成分,并评价其对肺癌细胞A549增殖、迁移的体外抑制作用。方法:采用水蒸气蒸馏法提取化州柚花挥发油;采用气质联用法(GC-MS)分析其化学组成,以峰面积归一化法计算各组分的相对百分含量;采用MTT法检测其对肺癌细胞A549增殖的抑制作用,计算细胞相对活力和增殖抑制率;采用细胞划痕实验评价其对肺癌细胞A549迁移能力的影响,计算相对迁移面积和迁移抑制率。结果:从100 g化州柚花中提取得到挥发油0.7 g,得率为0.7%。共从中分离得到了46个组分,鉴定了其中35个组分,占总峰面积的96.95%;相对百分含量较高的化合物包括D-柠檬烯(28.43%)、γ-萜品烯(20.74%)、β-月桂烯(11.97%)、芳樟醇(7.41%)等。与空白对照组比较,120 mg/L化州柚花挥发油可显著降低肺癌细胞A549的相对活力(P<0.05),增殖抑制率上升至21.72%;而1、5、10、20、40、60、80、100 mg/L化州柚花挥发油对肺癌细胞A549的相对活力均无显著影响(P>0.05)。作用24、48 h时,60、120 mg/L化州柚花挥发油可显著缩小肺癌细胞A549的相对迁移面积(P<0.05),迁移抑制率分别为132.46%、125.08%(24 h)和124.54%、124.66%(48 h);而30 mg/L化州柚花挥发油对肺癌细胞A549的相对迁移面积均无显著影响(P>0.05)。结论:GC-MS法可用于鉴定化州柚花挥发油的化学组成。化州柚花挥发油以D-柠檬烯、γ-萜品烯、β-月桂烯等成分为主,对肺癌细胞A549的增殖、迁移能力具有一定的体外抑制作用。

OBJECTIVE:To extract and identify the volatile oil from the flowers of Citrus grandis,and evaluate inhibitory effects of it on the proliferation and migration of lung cancer A549 cells in vitro.METHODS:The volatile oil was extracted from the flowers of C.grandis by steam distillation.The chemical constituents were analyzed by GC-MS.Relative percentage of each component was calculated by peak area normalization.Inhibitory effect of volatile oil on the proliferation of lung cancer A549 cells was detected by MTT assay,and the relative activity and proliferation inhibitory rate were calculated.Inhibitory effect of volatile oil on the migration of lung cancer A549 cells was evaluated by cell scratch test,and the relative migration area and migration inhibitory rate were calculated.RESULTS:Totally 0.7 g volatile oil was obtained from 100 g flowers of C.grandis with a yield of 0.7%.A total of 46 chemical components were isolated from the volatile oil,among which there were 35 compounds identified,accounting for 96.95%of the total peak area.The compounds with relatively higher percentage content included D-limonene(28.43%),γ-terpinene(20.74%),β-myrcene(11.97%)and linalool(7.41%),etc.Compared with blank control group,120 mg/L volatile oil from the flowers of C.grandis could significantly reduce the relative activity of lung cancer A549 cells(P<0.05),and the proliferation inhibitory rate increased to 21.72%.The volatile oil from the flowers of C.grandis had no obvious inhibitory effects on the relative activity of lung cancer A549 cells at the concentration of 1,5,10,20,40,60,80,100 mg/L(P>0.05).After treated for 24,48 h,the volatile oil from the flowers of C.grandis could significantly reduce the relative migration area of lung cancer A549 cells at the concentration of 60,120 mg/L(P<0.05);and the migration inhibitory rate were 132.46%,125.08%(24 h)and 124.54%,124.66%(48 h),respectively.But 30 mg/L volatile oil from the flowers of C.grandis had no obvious inhibitory effects on the relative migration area of lung cancer A549 cells(P>0.05).CONCLUSIONS:GC-MS can be used for identifying chemical compositions of volatile oil from the flowers of C.grandis.Volatile oil from the flowers of C.grandis are mainly D-limonene,γ-terpinene,β-myrcene,etc.,which can inhibit the the proliferation and migration of lung cancer A549 cells.