茅苍术麸炒品对人肝癌SMMC-7721细胞的体内外抑制作用  

Inhibitory effects of Atractylodes lancea(Thunb.)DC.stir-fried with bran on human hepatocellular carcinoma SMMC-7721 cells in vivo and in vitro

作　　者：罗吉辉[1] 杨熙华 肖方涛 周美华 LUO Ji-hui;YANG Xi-hua;XIAO Fang-tao;ZHOU Mei-hua(The Second Department of Tumor Surgery,South District of Chenzhou First People's Hospital,Chenzhou 423000,China)

机构地区：[1]湖南省郴州市第一人民医院南院肿瘤外科二区,郴州市423000

出　　处：《广西医学》2020年第11期1399-1403,1413,共6页Guangxi Medical Journal

基　　金：湖南省郴州市科技计划项目(2018cj102)。

摘　　要：目的探讨茅苍术麸炒品对人肝癌SMMC-7721细胞的体内外抑制作用。方法(1)取对数生长期SMMC-7721细胞,给予0μg/mL、25μg/mL、50μg/mL、100μg/mL、200μg/mL、400μg/mL和600μg/mL茅苍术麸炒品干预24 h后,检测SMMC-7721细胞的存活率;给予0μg/mL、100μg/mL、200μg/mL、400μg/mL茅苍术麸炒品干预24 h后,观察SMMC-7721细胞形态学,同时行Hoechst染色后于倒置荧光显微镜观察细胞凋亡情况,并应用流式细胞术检测细胞的凋亡率。(2)取24只裸鼠,皮下注射SMMC-7721细胞建立移植性肝癌模型。待肿瘤体积达到100 mm^3时,将裸鼠随机分为模型组和茅苍术麸炒品低、中、高剂量组,每组6只。低、中、高剂量组裸鼠分别每天灌胃0.8 g/kg、1.6 g/kg、3.2 g/kg的茅苍术麸炒品,模型组灌胃等体积的磷酸缓冲盐溶液。14 d后剥取肿瘤组织称质量,计算抑瘤率,并检测肿瘤组织B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关性X蛋白(Bax)和Survivin的蛋白表达水平。结果(1)100μg/mL、200μg/mL、400μg/mL和600μg/mL茅苍术麸炒品干预后细胞的存活率均低于0μg/mL,且随浓度增加细胞存活率依次下降(均P<0.05)。细胞形态学观察和Hoechst 33258染色可见,不同浓度茅苍术麸炒品干预后,细胞皱缩,并生成凋亡小体。0μg/mL、100μg/mL、200μg/mL、400μg/mL茅苍术麸炒品干预后细胞凋亡率依次升高(均P<0.05)。(2)模型组、茅苍术麸炒品低、中、高剂量组的肿瘤质量、Bcl-2和Survivin蛋白表达水平依次降低(均P<0.05),而抑瘤率、Bax蛋白表达水平均依次升高(均P<0.05)。结论茅苍术麸炒品能抑制肝癌SMMC-7721细胞增殖并诱导细胞凋亡,同时其能够抑制裸鼠移植瘤的生长,而这可能与其上调Bax蛋白表达和下调Bcl-2和Survivin蛋白表达有关。Objective To investigate the inhibitory effects of Atractylodes lancea(Thunb.)DC.stir-fried with bran on human hepatocellular carcinoma SMMC-7721 cells in vivo and in vitro.Methods(1)The SMMC-7721 cells at logarithmic growth phase were collected,then the survival rate of SMMC-7721 cells was detected after 24 hours of intervention with Atractylodes lancea(Thunb.)DC.stir-fried with bran of 0μg/mL,25μg/mL,50μg/mL,100μg/mL,200μg/mL,400μg/mL or 600μg/mL.After 24 hours of intervention with Atractylodes lancea(Thunb.)DC.stir-fried with bran of 0μg/mL,100μg/mL,200μg/mL or 400μg/mL,the morphology of SMMC-7721 cells was observed;moreover,cell apoptosis was observed under inverted fluorescence microscope after Hoechst staining,and cell apoptosis rate was measured by flow cytometry.(2)Twenty-four nude mice were selected and injected subcutaneously with SMMC-7721 cells for establishing a transplanted liver cancer model.The nude mice were randomized to model group,low-,moderate-and high-dose Atractylodes lancea(Thunb.)DC.stir-fried with bran groups when tumor volume reached 100 mm^3,with 6 mice in each group.Daily gavage administration of Atractylodes lancea(Thunb.)DC.stir-fried with bran was conducted in the low-,moderate-and high-dose Atractylodes lancea(Thunb.)DC.stir-fried with bran groups(0.8 g/kg,1.6 g/kg and 3.2 g/kg respectively),while intragastric administration of isometric phosphate-buffered saline was performed in the model group.After 14 days,the whole tumor tissues were removed and weighed up,then the antitumor rate was calculated;besides,the expression levels of B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax)and Survivin proteins in tumor tissues were determined.Results(1)The cell survival rate was decreased after intervention by Atractylodes lancea(Thunb.)DC.stir-fried with bran of 100μg/mL,200μg/mL,400μg/mL or 600μg/mL as compared with the rate after intervention with the concentration of 0μg/mL,and the cell survival rate declined with the increase of drug concentration(all P<0.05).Aft

关 键 词：肝癌 SMMC-7721细胞 裸鼠 移植瘤 茅苍术麸炒品 细胞增殖 细胞凋亡 B细胞淋巴瘤-2蛋白 B细胞淋巴瘤-2相关性X蛋白 Survivin蛋白 

分 类 号：R285.5[医药卫生—中药学] R735.7[医药卫生—中医学]