肉苁蓉总苷对HepG2细胞增殖、凋亡及Wnt/β-Catenin通路相关蛋白表达的影响  

Effects of Total Glycosides of Cistanche deserticola on Proliferation,Apoptosis and Expression of Wnt/β-Catenin Signaling Pathway Related Protein of HepG2 Cells

作　　者：冯朵 王靖 蒋勇军 周士琦 段昊 郭豫 赵建 闫文杰[1] FENG Duo;WANG Jing;JIANG Yongjun;ZHOU Shiqi;DUAN Hao;GUO Yu;ZHAO Jian;YAN Wenjie(Beijing Key Laboratory of Bioactive Substances and Functional Food,College of Biochemical Engineering,Beijing Union University,Beijing 100023,China;Institute of Food and Nutrition Development,Ministry of Agriculture and Rural Affairs,Beijing 100081,China;Inner Mongolia Sankou Biotechnology Co.,Ltd.,Ordos 017000,China)

机构地区：[1]北京联合大学生物化学工程学院,生物活性物质与功能食品北京市重点实验室,北京100023 [2]农业农村部食物与营养发展研究所,北京100081 [3]内蒙古三口生物科技有限公司,内蒙古鄂尔多斯017000

出　　处：《食品工业科技》2023年第20期389-397,共9页Science and Technology of Food Industry

基　　金：北京联合大学科研项目(No.ZKZD202303)。

摘　　要：为了探讨肉苁蓉总苷(total glycosides of Cistanche deserticola,TG)对HepG2细胞的抑制作用及其机制研究。采用不同浓度(0、3.5、10.5、21、31.5、42μg/mL)TG处理HepG2细胞24 h后,使用CCK8法检测细胞存活率;应用Hoechst 33342/PI双染法及Annexin V-FITC/PI检测细胞凋亡;并通过细胞迁移试验检测细胞迁移现象;同时采用流式细胞仪检测细胞周期进展变化;通过Western blot法测定α-甲胎蛋白(α-fetoprotein,AFP)、β-连环蛋白(β-catenin)、蓬乱蛋白(Dishevelled,Dsh)、糖原合成酶激酶-3β(GSK-3β)的表达量。结果发现,TG可以降低HepG2细胞的增殖能力,且有浓度依赖性,当TG为42μg/mL时,细胞存活率仅有31.04%;此外,TG可以破坏细胞结构,诱导细胞凋亡,AV/PI检测后细胞凋亡率可高达32.44%;还可促使细胞坏死,限制细胞迁移,处理后的组别与对照组之间存在显著差异(P<0.05或P<0.01);同时,高浓度TG可以阻滞HepG2细胞在S期和G2/M期;与对照组相比,TG处理之后,β-catenin、Dsh表达下降,而GSK-3β表达量升高。由此可知,肉苁蓉总苷通过影响细胞周期进展、促进细胞凋亡、限制细胞迁移等方面抑制HepG2细胞增殖,其作用机制可能是通过Wnt/β-catenin信号通路,激活GSK-3β降解β-catenin来实现肝癌抑制作用的。To explore the inhibitory effect of total glycosides of Cistanche deserticola(TG)on HepG2 cells and its mechanism.In this paper,different concentrations(0,3.5,10.5,21,31.5,42μg/mL)TG were treated 24 h on HepG2 liver cancer cells,and the viability of HepG2 cells was detected using CCK8 assay.Hoechst 33342/PI double staining method and Annexin V-FITC/PI were used to detect HepG2 cells apoptosis.The phenomenon of cell migration was detected by cell migration assay.Meanwhile,cell cycle progression changes were detected by flow cytometry.And the expression of α-fetoprotein(AFP),β-catenin,Dishevelled(Dsh),GSK-3β were detected by Western blot.The results showed that TG could reduce the proliferation of HepG2 cells in a concentration dependent manner,with only 31.04% cell viability when TG was 42μg/mL.In addition,TG could damage the cell structure and induce cell apoptosis,and the apoptosis rate could be as high as 32.44% by AV/PI detection.Moreover,TG could also promote cell necrosis,and limit cell migration.There were significant differences between the treated group and the control group(P<0.05 or P<0.01).Meanwhile,high concentration of TG could arrest HepG2 cells in S and G2/M phase.Finally,compared with the control group,the relative expression of β-catenin and Dsh decreased,while GSK-3β increased in the TG treated groups.In conclusion,TG could inhibit HepG2 cell proliferation by affecting cell cycle progression,promoting apoptosis,and limiting cell migration.The mechanism might be through Wnt/β-catenin signaling pathway,which activated GSK-3β to degrade β-catenin to achieve liver cancer inhibition.

关 键 词：荒漠肉苁蓉 肉苁蓉总苷 HEPG2细胞 抗肝癌 WNT/Β-CATENIN通路 

分 类 号：R284.2[医药卫生—中药学]