【摘要】 目的体外观察中药仙鹤草的抗肝癌作用,并初步探讨其机制。方法制备仙鹤草水提液,以含不同浓度仙鹤草水提液的细胞培养液培养SMMC-7721肝癌细胞,于不同时间点采用MTT法测定细胞活力并计算抑制率(IR),光镜下观察细胞形态学变化,流式细胞仪检测细胞凋亡率,免疫细胞化学染色检测Bcl-2、P53蛋白的表达。结果仙鹤草水提液作用SMMC-7721肝癌细胞24、48、72h后,5mg/mL组和10mg/mL组无明显细胞生长抑制作用,20mg/mL组的IR分别为0.5%、23.9%和27.5%,40mg/mL组的IR分别为23.3%、51.7%和71.6%。仙鹤草水提液有效浓度作用48、72h后,细胞呈现明显的凋亡形态学改变;20mg/mL组的细胞凋亡率分别为19.5%和23.0%,40mg/mL组分别为33.4%和42.7%。20mg/mL组的Bcl-2蛋白阳性表达率分别为71.9%和58.5%,40mg/mL组分别为47.9%和26.5%。20mg/mL组的P53蛋白阳性表达率分别为22.9%和50.6%,40mg/mL组分别为48.7%和83.7%。结论仙鹤草水提液能较明显地抑制SMMC-7721肝癌细胞增殖,诱导细胞凋亡,并呈剂量、时间依赖性,凋亡发生可能与Bcl-2和P53蛋白表达改变有关。更多还原

【Abstract】 Objective To observe the anti-tumor effects of Agrimonia Pilosa Ledeb(APL),a Chinese herbal medicine,on hepatocellular carcinoma cells in vitro and investigate the underlined mechanisms preliminarily.Methods APL water extracts were prepared.SMMC-7721 cells were cultured with the medium containing different concentrations of APL water extracts,and at different time points,cell viabilities were measured by the MTT assay and inhibitory rates(IR)were calculated;cell morphologic changes were observed under a light microscope;apoptotic ratios were measured by flow cytometry;and the expressions of Bcl-2 and P53 proteins were examined by immunocytochemistry.Results After the cells were cultured with the medium containing APL water extracts for 24h,48h and 72h,no obvious effects were found on the cell proliferation in 5mg/mL group and 10mg/mL group,but IR were 0.5%,23.9% and 27.5% in 20mg/mL group and 23.3%,51.7% and 71.6% in the 40mg/mL group,respectively.In the groups with effects on the cells proliferation,morphological characteristics of apoptosis were obvious,and the cell apoptotic ratios were 19.5%and 23.0% in 20mg/mL group and 33.4% and 42.7% in 40mg/mL group at 48h and 72h.The expressions of Bcl-2 protein were 71.9%and 58.5% in 20mg/mL group and 47.9% and 26.5% in 40mg/mL group at 48h and 72h,and the expressions of P53 protein were 22.9% and 50.6% in 20mg/mL group and 48.7% and 83.7% in 40mg/mL group at 48hand 72h.Conclusion The water extracts of APL are able to inhibit proliferation and induce apoptosis of SMMC-7721 cells dose-time dependently in vitro,which might be associated with the expression changes of Bcl-2 and P53 protein.更多还原