酒糟粗提物对肝癌细胞HepG2增殖的抑制作用  

Inhibition on the Growth of HepG2 Cells by Crude Extraction from Distillers'Grains

作　　者：王若帆 侯茂[3] 谢梦忆[1,2] 游川 李敬东[1,2] WANG Ruofan;HOU Mao;XIE Mengyi;YOU Chuan;LI Jingdong(Department of Hepatobiliary Surgery,Affiliated Hospital of North Sichuan Medical College,Nanchong 637200,China;Institute of Hepato-Biliary-Pancreas and Intestinal Disease,North Sichuan Medical College,Nanchong 637200,China;Key Laboratory of Sichuan Province for the Utilization of Solid State Fermentation Resources,Yibin University,Yibin 644000,China)

机构地区：[1]川北医学院附属医院肝胆外科,四川南充637200 [2]川北医学院肝胆胰肠疾病研究所,四川南充637200 [3]宜宾学院固态发酵资源利用四川省重点实验室,四川宜宾644000

出　　处：《食品工业科技》2021年第21期392-399,共8页Science and Technology of Food Industry

基　　金：中国轻工业浓香型白酒固态发酵重点实验室开放基金(2017JJ008)。

摘　　要：为了解酒糟粗提物对人体肝癌细胞株HepG2细胞增殖和凋亡的影响,首先采用液相色谱-质谱联用仪(LCMS)分析酒糟粗提物成分,再利用不同浓度的酒糟提取物分别处理HepG2细胞,采用RTCA检测其对HepG2细胞增殖的影响,采用流式细胞术检测其对HepG2细胞周期的影响,采用实时荧光定量PCR检测其对CyclinA、CDK2、Bax、Bcl-2基因表达的影响,采用Western Blot检测其对9种凋亡相关蛋白表达量的影响。研究发现浓香型白酒酒糟粗提物主要含有43种物质,酒糟粗提物可明显抑制HepG2细胞的增殖且呈浓度依赖;酒糟粗提物可以显著下调S期相关周期蛋白CyclinA及其激酶CDK2的mRNA及蛋白的表达,同时使抗凋亡蛋白Bcl-2的mRNA及蛋白的表达减少,促凋亡蛋白Bax的mRNA及蛋白的表达增加(P<0.05);CYTC、Cleaved-Caspase-9及Cleaved-Caspase-3表达量逐渐升高(P<0.05),Caspase-9、Caspase-3表达量逐渐降低(P<0.05)。结果表明酒糟粗提物含多种活性物质,可抑制HepG2细胞的增殖,并通过激活介导凋亡的线粒体通路诱导肝癌HepG2细胞发生凋亡。To explore the effect of the crude extraction from distiller's grains on the proliferation and apoptosis of human hepatocellular carcinoma cell HepG2,the crude extraction from distiller's grains were analyzed by LC-MS firstly,then used to treat HepG2 cells by different concentrations.RTCA was used to detect its effects on proliferation of HepG2 cells.Flow cytometry was used to detect its effects on the cycle of HepG2 cells.Real-Time qPCR was used to detect its effects on apoptosis of HepG2 cells.Western Blot was used to detect the expression of apoptotic-related protein.43 substances in the crude extraction from distiller's grains were determined by LC-MS.The proliferation of HepG2 cells were significantly inhibited by the crude extraction from distiller's grains and presented concentration dependent;The crude extraction from distiller's grains significantly down-regulated the expression of mRNA and protein of S phase CyclinA and its kinase CDK2,reduced the expression of mRNA and protein of Bcl-2 and increased the expression of mRNA and protein of Bax(P<0.05);the expression levels of CYTC,Cleaved-Caspase-9 and Cleaved-Caspase-3 were increased gradually(P<0.05),the expression levels of Caspase-9,Caspase-3 were decreased(P<0.05).The results showed that the crude extraction from distiller's grains that contained a variety of active substances can significantly inhibit the proliferation of HepG2 cells and may induce apoptosis of HepG2 cells by activating the apoptotic mitochondrial pathway.

关 键 词：酒糟粗提物 成分 HEPG2 细胞 抑制 细胞周期 凋亡蛋白 

分 类 号：TS201.1[轻工技术与工程—食品科学]