刺头复叶耳蕨总提取物与总黄酮体外抗肿瘤活性的比较研究

Comparison of Anti-tumor Activities of Total Flavones and Extracts from Arachniodes Exilis(Hance) Ching in Vitro

关键词:  刺头复叶耳蕨  总黄酮  肿瘤细胞  增殖

Key words:  Arachniodes exilis(Hance) Ching  total flavones  tumor cells  proliferation

摘要:

目的 比较刺头复叶耳蕨总提取物与总黄酮体外抗肿瘤活性。方法 以不同浓度的刺头复叶耳蕨总提取物与刺头复叶耳蕨总黄酮分别作用于人肝癌HepG2细胞、人肺癌A549细胞和人成骨肉瘤Saos2细胞，400 nmol·L^-1槲皮素作阳性对照，0.1% DMSO的完全培养基为阴性对照，倒置显微镜下连续观察3 d各组细胞形态及密度的变化；CCK-8法检测药物的抑制率及半数抑制浓度(IC₅₀)，流式细胞仪检测HepG2细胞凋亡情况。结果 与阴性对照组比较，人肝癌HepG2细胞和肺癌A549细胞加入不同终浓度复叶耳蕨总提取物与总黄酮，1 d后发现细胞生长出现明显抑制，细胞收缩变圆、裂解，且呈浓度依赖，而人骨肉瘤Saos2细胞形态无明显改变。CCK-8法检测显示复叶耳蕨总提物、总黄酮对人肝癌HepG2细胞和肺癌A549细胞的抑制呈时间及浓度依赖关系，复叶耳蕨总提物和复叶耳蕨总黄酮对HepG2作用的IC₅₀分别为76.62，36.75 μg·mL^-1，对A549作用的IC50分别是170.13，117.60 μg·mL^-1。一定浓度刺头复叶耳蕨总黄酮可引起HepG2细胞凋亡。结论 刺头复叶耳蕨总黄酮体外抗肿瘤活性优于总提取物。

Abstract:

OBJECTIVE To compare the anti-tumor activities of total flavones and extracts from Arachniodes Exilis(Hance) Ching in vitro. METHODS Total extracts and flavones from Arachniodes exilis(Hance) Ching in different concentration were respectively used on human hepatocellular carcinoma HepG2 cells, human lung cancer A549 cells and osteosarcoma Saos2 cells. Positive control was 400 nmol·L^-1 quercetin, and negative control was 0.1% DMSO complete medium. The changes of shape and density of cells were observed continuously in inverted phase contrast microscope for 3 d. CCK-8 detection was used to detect inhibition ratio and half maximal inhibitory concentration(IC₅₀). Apoptosis of HepG2 cells were detected by flow cytometry. RESULTS Compared with the negative control group, the growth of A549 cells and HepG2 cells in different concentration of total extracts and flavones were inhibited after 24 h. The cells shrinked, turned round and cracked with concentration dependence. However, Saos-2 cells did not show obviously morphological changes. The result of CCK-8 detection suggested that the inhibition of total extracts and flavones on HepG2 cells and A549 cells had a concentration and time dependence. The IC₅₀ of total extracts and flavones on HepG2 cells were 76.62, 36.75 μg·mL^-1 and 170.13, 117.60 μg·mL^-1 on A549 cells. Certain concentration of flavones induced apoptosis of HepG2 cells. CONCLUSION The anti-tumor activity of total flavones from Arachniodes exilis(Hance) Ching is better than total extracts.