蒲葵子乙醇提取物调控糖酵解相关基因和蛋白抑制Huh-7细胞增殖的研究

doi:10.3760/cma.j.cn441417-20250430-21006

国际医药卫生导报 ›› 2025, Vol. 31 ›› Issue (21): 3545-3550.DOI: 10.3760/cma.j.cn441417-20250430-21006

蒲葵子乙醇提取物调控糖酵解相关基因和蛋白抑制Huh-7细胞增殖的研究

李悦轩¹ 张晓源² 陈淑游² 孙瑞涵² 陈旭征²^，³ 林薇⁴

¹福建中医药大学药学院，福州 350122；²福建中医药大学中西医结合学院中西医结合研究院，福州 350122；³福建省中西医结合老年性疾病重点实验室，福州 350122；⁴福建中医药大学科技创新与转化中心，福州 350122

收稿日期:2025-04-30 出版日期:2025-11-01 发布日期:2025-11-18
通讯作者: 林薇，Email：linwei311616@126.com
基金资助:
福建省自然科学基金（2019J01727）

Study on the inhibitory effect of ethanol extract of Livistona chinensis on the proliferation of Huh-7 cells by regulating glycolysis related genes and proteins

Li Yuexuan¹, Zhang Xiaoyuan², Chen Shuyou², Sun Ruihan², Chen Xuzheng^2,3, Lin Wei⁴

¹School of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China; ²School of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China; ³Fujian Key Laboratory of Integrative Medicine on Geriatrics, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China; ⁴Center for Scientific Innovation and Translation, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China

Received:2025-04-30 Online:2025-11-01 Published:2025-11-18
Contact: Lin Wei, Email: linwei311616@126.com
Supported by:
Fujian Natural Science Foundation (2019J01727)

摘要/Abstract

摘要： 目的　探究蒲葵子乙醇提取物（EELC）抑制肝癌Huh-7细胞增殖的作用是否与其调控糖酵解过程相关。方法　本实验于2022年1月至2023年12月在福建省中西医结合老年性疾病重点实验室完成。运用MTT法评估经不同浓度EELC处理24 h后对Huh-7细胞活力的影响；通过Seahorse XFe 96细胞能量代谢分析仪，对线粒体ATP产生速率和糖酵解ATP产生速率进行实时动态监测，并同步测定实时耗氧率、胞外酸化速率及质子流出速率的瞬时变化；采用Real-time PCR和Western blot分别检测葡萄糖转运蛋白（Glut1、Glut3）、己糖激酶（HK）、缺氧诱导因子1α（HIF-1α）的mRNA和蛋白表达水平。多组间比较采用单因素方差分析，重复测量资料采用单因素重复测量方差分析。结果　EELC呈浓度依赖性抑制Huh-7细胞增殖。细胞能量代谢分析显示，EELC能提高线粒体ATP产生速率，同时降低糖酵解ATP产生速率。EELC处理组的实时耗氧率明显增加，胞外酸化速率和质子流出速率均显著降低（均P<0.000 1）。分子水平检测发现，EELC能下调HIF-1α的mRNA和蛋白表达水平，并降低Glut1蛋白表达水平（均P<0.01）。结论　EELC通过调节细胞能量代谢、HIF-1α的mRNA表达水平和糖酵解相关蛋白HIF-1α、Glut1的表达水平，降低糖酵解能力，并上调线粒体功能以增强细胞的有氧呼吸能力，从而有效抑制Huh-7细胞的增殖与生长。

关键词: 肝细胞癌, Huh-7细胞, 糖酵解, 蒲葵子乙醇提取物

Abstract: Objective　To investigate whether the inhibitory effect of ethanol extracts from Livistona chinensis seeds (EELC) on the proliferation of Huh-7 hepatocellular carcinoma cells is related to its regulation of the glycolytic process. Methods　This experiment was conducted from January 2022 to December 2023 at the Key Laboratory of Geriatric Diseases of Integrated Traditional Chinese and Western Medicine, Fujian Province. The MTT assay was used to evaluate the effects of different concentrations of EELC on Huh-7 cell viability after 24 hours of treatment. The Seahorse XFe 96 Analyzer was employed for real-time dynamic monitoring of mitochondrial ATP production rates and glycolytic ATP production rates, while simultaneously measuring instantaneous changes in oxygen consumption rate, extracellular acidification rate, and proton efflux rate. Real-time PCR and Western blotting were utilized to detect the mRNA and protein expression levels of hypoxia-inducible factor 1-alpha (HIF-1α), hexokinase (HK), and glucose transporters (Glut1, Glut3). One-way analysis of variance was used for comparison among multiple groups, and one-way repeated measurement analysis of variance was used for repeated measurement data. Results　EELC exerted a concentration-dependent inhibitory effect on Huh-7 cell proliferation. Cell energy metabolism analysis showed that EELC could increase mitochondrial ATP production rate and reduce glycolysis ATP production rate. The real-time oxygen consumption rate of the EELC treatment group was significantly increased, and the extracellular acidification rate and proton outflow rate were significantly decreased (all P<0.0001). Molecular level assessments indicated that EELC downregulated the mRNA and protein expression of HIF-1α and reduced Glut1 protein expression (all P<0.01). Conclusions　EELC effectively inhibits the proliferation and growth of Huh-7 cells by regulating cellular energy metabolism, downregulating HIF-1α mRNA levels, and reducing the expression of glycolysis-related proteins HIF-1α and Glut1. This results in decreased glycolytic capacity and enhanced mitochondrial function to improve cellular aerobic respiration.

Key words: Hepatocellular carcinoma, Huh-7 cells, Glycolysis, Ethanol extract of Livistona chinensis

李悦轩张晓源陈淑游孙瑞涵陈旭征林薇. 蒲葵子乙醇提取物调控糖酵解相关基因和蛋白抑制Huh-7细胞增殖的研究[J]. 国际医药卫生导报, 2025, 31(21): 3545-3550.

Li Yuexuan, Zhang Xiaoyuan, Chen Shuyou, Sun Ruihan, Chen Xuzheng, Lin Wei. Study on the inhibitory effect of ethanol extract of Livistona chinensis on the proliferation of Huh-7 cells by regulating glycolysis related genes and proteins[J]. International Medicine and Health Guidance News, 2025, 31(21): 3545-3550.

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蒲葵子乙醇提取物调控糖酵解相关基因和蛋白抑制Huh-7细胞增殖的研究

Study on the inhibitory effect of ethanol extract of Livistona chinensis on the proliferation of Huh-7 cells by regulating glycolysis related genes and proteins

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