CAJ | 학술논문

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、72 h后，5 mg/mL组和10 mg/mL 组无明显细胞生长抑制作用，20 mg/mL 组的 IR分别为0．5％、23．9％和27．5％，40 mg/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、72 h后，细胞呈现明显的凋亡形态学改变；20 mg/mL组的细胞凋亡率分别为19．5％和23．0％，40 mg/mL组分别为33．4％和42．7％。20 mg/mL组的Bcl-2蛋白阳性表达率分别为71．9％和58．5％，40 mg/mL组分别为47．9％和26．5％。20 mg/mL组的P53蛋白阳性表达率分别为22．9％和50．6％，40 mg/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。
목적：체외관찰중약선학초적항간암작용，병초보탐토기궤제。방법제비선학초수제액，이함불동농도선학초수제액적세포배양액배양SMMC-7721간암세포，우불동시간점채용MTT법측정세포활력병계산억제솔（IR），광경하관찰세포형태학변화，류식세포의검측세포조망솔，면역세포화학염색검측Bcl-2、P53단백적표체。결과선학초수제액작용SMMC-7721간암세포24、48、72 h후，5 mg/mL조화10 mg/mL 조무명현세포생장억제작용，20 mg/mL 조적 IR분별위0．5％、23．9％화27．5％，40 mg/mL조적IR분별위23．3％、51．7％화71．6％。선학초수제액유효농도작용48、72 h후，세포정현명현적조망형태학개변；20 mg/mL조적세포조망솔분별위19．5％화23．0％，40 mg/mL조분별위33．4％화42．7％。20 mg/mL조적Bcl-2단백양성표체솔분별위71．9％화58．5％，40 mg/mL조분별위47．9％화26．5％。20 mg/mL조적P53단백양성표체솔분별위22．9％화50．6％，40 mg/mL조분별위48．7％화83．7％。결론선학초수제액능교명현지억제SMMC-7721간암세포증식，유도세포조망，병정제량、시간의뢰성，조망발생가능여Bcl-2화P53단백표체개변유관。
Objective To observe the anti-tumor effects of Agrimonia Pilosa Ledeb(APL) ,a Chinese herbal medicine ,on hepato-cellular carcinoma cells in vitro and investigate the underlined mechanisms preliminarily .Methods APL water extracts were pre-pared .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 ex-tracts for 24 h ,48 h and 72 h ,no obvious effects were found on the cell proliferation in 5 mg/mL group and 10 mg/mL group ,but IR were 0 .5% ,23 .9% and 27 .5% in 20 mg/mL group and 23 .3% ,51 .7% and 71 .6% in the 40 mg/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 20 mg/mL group and 33 .4% and 42 .7% in 40 mg/mL group at 48 h and 72 h .The expressions of Bcl-2 protein were 71 .9% and 58 .5% in 20 mg/mL group and 47 .9% and 26 .5% in 40 mg/mL group at 48 h and 72 h ,and the ex-pressions of P53 protein were 22 .9% and 50 .6% in 20 mg/mL group and 48 .7% and 83 .7% in 40 mg/mL group at 48 h and 72 h . Conclusion The water extracts of APL are able to inhibit proliferation and induce apoptosis of SMMC-7721 cells dose-time depend-ently in vitro ,which might be associated with the expression changes of Bcl-2 and P53 protein .