槐米中天然产物槲皮素抗人鼻咽癌CNE1细胞活性研究

本研究以槐米为原材料提纯天然产物槲皮素,以人鼻咽癌细胞系CNE1为试验对象,探索天然产物槲皮素对人鼻咽癌细胞的增殖抑制效应及凋亡诱导效应。研究方法采用超声醇提法从槐米中提取芸香苷,再经酸水解和重结晶制备槲皮素精制品;采用槲皮素标准曲线比色法检测槲皮素精制品中槲皮素的纯度;采用MTT比色法进行细胞毒性试验,检测槲皮素精制品对人鼻咽癌细胞CNE1增殖的抑制效应及半数抑制浓度(IC50);采用流式细胞术结合AnnexinV、PI双染色法进行细胞凋亡试验,检测槲皮素精制品对CNE1细胞凋亡的诱导效应。结果表明,从槐米中提纯的天然产物槲皮素精制品,能呈剂量依赖性地抑制人鼻咽癌细胞CNE1增殖并诱导其凋亡。综上所述,应用超声醇提法,从槐米中制备的天然产物槲皮素精制品,不仅纯度高,且有着较优的抗癌活性,进一步明确了槐米等中药材开发天然产物有效成分,服务于人类健康的潜在应用价值和社会与经济效益。

Study on Natural Product of Quercetin from Flos Sophorae Immaturus against Activity of Human Nasopharyngeal Carcinoma CNE1 cells

In this study,the effects of quercetin from Sophora japonica buds were investigated on proliferation(inhibitory effect)and apoptosis(induction effect)of Nasopharyngeal carcinoma cell CNE1,basing on the natural product quercetin purified from the raw material Sophora japonica buds.Rutin was prepared from Flos Sophorae Immaturus with ultrasonic assisted and ethanol extraction method and then the purified product of quercetin was obtained by following acid hydrolysis and recrystallization.The purity of quercetin was detected with quercetin standard curve by colorimetric method.Cytotoxicity test was carried out by MTT assay,including the inhibitory effect of quercetin on cell proliferation in human nasopharyngeal carcinoma CNE1 cells and the half inhibition concentration(IC50).The induction effect of quercetin on cell apoptosis of human nasopharyngeal carcinoma cell line CNE1 was measured for the cell apoptosis test by flow cytometry(FCM)labeled with Annexin V/PI.The results show that the natural products of quercetin purified from in Flos Sophorae Immaturus can inhibit proliferation in human nasopharyngeal carcinoma CNE1 cells with a dose-dependent manner and induces apoptosis.In conclusion,the natural product of quercetin,prepared with the ultrasonic assisted method and ethanol extraction method,was not only high purity but also processing a good anti-tumor activity.In addition,the effective components of natural products by Flos Sophorae Immaturus and other Chinese medicinal materials were further supported and clear,which would have potential application values and economic benefits for human health.