瑶药苦赖咪总醌提取物的制备及其抗肝癌活性研究  

Preparation of total quinone extract from traditional Yao drug Rumex daiwoo Makino and its activity against hepatoma

作　　者：蒙永 龙奕霄[1,2] 蒋霞 MENG Yong;LONG Yi-xiao;JIANG Xia(School of Pharmacy,Guangxi Medical University,Nanning 530021,China;Department of Pharmacy,Wuzhou Red Cross Hospital,Wuzhou 543002,China;Department of Pharmacy,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学药学院,南宁市530021 [2]广西梧州红十字会医院药学部,梧州市543002 [3]广西医科大学第一附属医院药学部,南宁市530021

出　　处：《广西医学》2022年第2期179-183,共5页Guangxi Medical Journal

基　　金：广西中医药适宜技术开发与推广项目(GZSY20-09);广西壮族自治区卫生健康委员会自筹经费科研课题(Z20190913,Z20190916)。

摘　　要：目的研究瑶药苦赖咪总醌提取物的制备方法,并分析该提取物对人肝癌细胞的抑制作用。方法采用醇提再酸碱处理及有机溶剂萃取的方法获得瑶药苦赖咪的醌类提取物,采用分光光度法对提取物中总醌的含量进行测定。使用不同浓度苦赖咪总醌提取物干预人肝癌HepG2细胞后,采用细胞计数检测法检测细胞增殖抑制率,通过细胞克隆形成实验、划痕实验分别检测HepG2细胞克隆、迁移能力,并采用PCR法检测HepG2细胞中肿瘤坏死因子α(TNF-α)、白细胞介素8(IL-8)mRNA的表达水平。结果(1)提取物的总醌含量为52.31%。(2)随着干预浓度的增加,苦赖咪总醌提取物对HepG2细胞的抑制作用总体呈增强趋势,且在25~200μg/mL的浓度范围内其抑制作用呈浓度依赖性。(3)经50μg/mL、100μg/mL苦赖咪总醌提取物干预24 h后,HepG2细胞的克隆形成数减少,细胞迁移率降低,且药物浓度越高,细胞克隆形成数越少,细胞迁移率越低(均P<0.05)。(4)经100μg/mL苦赖咪总醌提取物干预后HepG2细胞中的TNF-αmRNA表达水平升高,而IL-8 mRNA表达水平下降(均P<0.05)。结论瑶药苦赖咪的总醌提取物能够抑制人肝癌细胞增殖、克隆和迁移能力,其可能通过上调TNF-α表达、下调IL-8表达而发挥抗肿瘤作用。Objective To study the preparing approach of the total quinone extract from traditional Yao drug Rumex daiwoo Makino,and to analyze the inhibitory effect of this extract on human hepatoma cells.Methods The quinones extract from traditional Yao drug,Rumex daiwoo Makino,were obtained using ethanol extraction followed by acid-alkali treatment and the organic solvent extraction method.The content of total quinone in the extracts was determined by spectrophotometry.After the interventions on human hepatoma HepG2 cells with different concentrations of the total quinone extract from Rumex daiwoo Makino,the inhibition rate of cell proliferation was detected by cell counting kit-8 assay,and the cloning and migration abilities of HepG2 cells were detected by cell colony forming assay and wound-healing assay,respectively.Besides,PCR assay was used to detect the expression levels of tumor necrosis factorα(TNF-α)and interleukin 8(IL-8)mRNAs in HepG2 cells.Results(1)The content of total quinone in the extract was 52.31%.(2)Overall,the inhibitory effect of the total quinone extract from Rumex daiwoo Makino on HepG2 cells increased with the growing intervention concentration,and the extract with a concentration between 25 and 200μg/mL exerted an inhibitory effect in a dose-dependent manner.(3)After a 24-hour intervention with the total quinone extract from Rumex daiwoo Makino(50 or 100μg/mL),HepG2 cells exhibited a decreased number of colonies and a reduced cell migration rate;moreover,the number of cell colonies and the cell migration rate declined with the increasing concentration of drug(all P<0.05).(4)After the intervention with the total quinone extract from Rumex daiwoo Makino(100μg/mL),the expression level of TNF-α mRNA was increased while the expression level of IL-8 mRNA was decreased in HepG2 cells(all P<0.05).Conclusion The total quinone extract of traditional Yao drug,Rumex daiwoo Makino,can inhibit the proliferation,cloning and migration abilities of human hepatoma cells,which may exert antitumor effects by up

关 键 词：肝癌 苦赖咪 总醌提取物 抗肿瘤作用 炎症因子 瑶药 体外实验 

分 类 号：R29[医药卫生—民族医学] R273[医药卫生—中医学]