鄂西香茶菜中4个二萜类成分的含量及其对细胞增殖的影响

Contents of Four Diterpenoids and Cytotoxic Activity in Isodon henryi

目的:以冬凌草甲素,毛叶香茶菜素E,毛栲利素,香茶菜素B为指标,建立HPLC同时测定鄂西香茶菜中4个二萜的质量控制方法,并对4个成分进行体外抗肿瘤活性的评价。方法:采用YMC C18反相色谱柱(4.6 mm×250 mm,5μm),流动相乙腈-水(38∶62),流速1.0 m L·min^(-1),柱温30℃,检测波长230 nm;采用噻唑蓝(MTT)比色法测定了鄂西香茶菜中4个指标性成分对HCT^(-1)16,Hep G2,BGC-823,MB-231,A2780细胞的增殖抑制作用。结果:冬凌草甲素,毛叶香茶菜素E,毛栲利素,香茶菜素B分别在0.085 6~0.856,0.014 04~0.140 4,0.070 8~0.708,0.018 98~0.189 8 g·L^(-1)呈良好的线性关系,平均回收率分别为101.78%,101.70%,100.23%,104.60%。4个二萜均显示了一定的细胞增殖抑制作用,其中冬凌草甲素,毛栲利素对5种细胞株的增值抑制率均>50%。冬凌草甲素、毛栲利素对5株肿瘤细胞株的细胞毒作用均优于毛叶香茶菜素E和香茶菜素B。半数抑制浓度(IC50)分别为4.10~6.46,2.34~4.77μmol·L^(-1)。结论:该方法简便,灵敏,重复性好,可用于鄂西香茶菜中主要细胞增殖抑制二萜的质量控制。

Objective： To establish a method for simultaneous determination of four diterpenoids in Isodon henryi, and evaluate the in vitro antitumor activity of four diterpenes （oridonin, zea pollinium, lasiokaurin, rabdosin B）. Method： YMC C18 column （4.6 mm × 250 mm, 5 μm） was adopted with acetonitrile-water （38：62） as mobile phase for isocratic elution. The flow rate was 1.0 mL min-1 ; the column temperature was 30 ℃ ; and the detection wavelength was at 230 nm. They were tested for their cytotoxic effects against five human cancer cell lines： HCT-116, HepG2, BGC-823, MB-231, and A2780 by MTT. Result： A good linearity wasobserved in the range of 0.085 6-0.856, 0.014 04-0. 140 4, 0.070 8-0.708, 0.018 98-0. 189 8 g-L-1 for oridonin, maoyecrystal E, lasiokaurin, rabdosin B, with the average recoveries of 101.78%, 101.70%, 100.23% , 104.60% , respectively; 4 diterpenes showed a certain cytotoxie effect, in which oridonin, lasiokaurin 5 kinds of cell strains of the rate of inhibition were greater than 50%. Oridonin, lasiokaurin exhibited more obviously cytotoxic activity in the tested cell lines with IC50 values ranging from 4. 10-6.46, 2. 34-4.77 μmmol·L-1 respectively. Conclusion： The method is simple, sensitive and reproducible, and it can be used for the quality control of the main cytotoxic active diterpenes in L henryi.