青龙衣提取物对人肝癌细胞株抑制作用的实验研究

摘要

目的：通过青龙衣提取物对人肝癌细胞株SMMC-7721的抑制作用，筛选出其抑瘤活性成分并探讨其机制。方法：青龙衣抑瘤活性成分胡桃醌与氟尿嘧啶、奥沙利铂、亚砷酸注射液分别作用于人肝癌细胞株SMMC-7721，分别于24h、48h、72h收集细胞，采用MTT法比较胡桃醌与其他组药物的抑瘤率，借助电镜观察胡桃醌作用于人肝癌细胞株SMMC-7721后细胞微细结构的形态变化，于激光共聚焦显微镜下检测细胞内Ca2＋浓度变化。结果：MTT结果显示胡桃醌抑瘤具有剂量依赖性；0．01mg／mL剂量24、48 hMTT结果显示胡桃醌的抑瘤强度高于亚砷酸注射液，与氟尿嘧啶相当，低于奥沙利铂；电镜结果显示肝癌细胞染色质凝聚，核固缩，游离核糖体减少，出现大溶酶体颗粒，线粒体数量增多形成髓样变，部分细胞坏死；激光共聚焦显微镜检测结果显示胡桃醌作用于肝癌细胞SMMC-7721后细胞内Ca2＋浓度增加。结论：青龙衣提取物抑瘤活性成分为化合物胡桃醌；胡桃醌对人肝癌细胞株SMMC-7721的抑制作用可能是直接杀伤肿瘤细胞。%Objective:By means of inhibition of Qinglongyi extracts to hepatocellular carcinoma cell,screen out its antitu-mor activity and mechanism of ingredients.Methods: Juglone was the anti-tumor active ingredient of Qinglongyi and fluorouracil, Oxaliplatin, Arsenic trioxide injection were acted on the human hepatocellular carcinoma cell line SMMC-7721, respectively, for 24h, 48h, 72h.The cells were collected by MTT assay compared with other groups of drugs on the inhibition rate.Electron microscope was used to observe the changes of fine structure of hepatocellular carcinoma cell line SMMC-7721 cells treated by juglone.Intracellular Ca2+concentration was detected with Laser scanning confocal microscope.Results:MTT results showed a dose-dependent inhibition of juglone;MTT results at the dose of 0.01mg/ml in 24h, 48h showed that Juglone’s antitumor intensity was higher than arsenite injection,the same with oxaliplatin, and lower than fluorouracil.The results of electron microscope revealed chromatin condensation,karyopyknosis,reduction of free ribosomes,big lysosomal granules,forming of medullary change by increasing mitochondria,and partial necrosis in hepatocellular carcinoma cell.Laser scanning confocal microscopy showed increasesing of intracellular Ca2+concentration in hepatocellular carcinoma cells SMMC-7721 after treating by juglone.Conclusion:The anti-tumor active ingredient of Qinglongyi was the compound juglone.The inhibitory effect of juglone on human hepatoma cell line SMMC-7721 may be shown by directly killing tumor cells.