芪灵扶正清解方对Huh-7细胞能量代谢及糖酵解相关蛋白的影响  

Effect of Qiling Fuzheng Qingjie Formula on Energy Metabolism and Glycolysis Related Proteins in Huh-7 Cells

作　　者：颜硕 刘海琴 郭康悦 何家珺 曹治云 章尤权[5] 林明和 陈旭征 YAN Shuo;LIU Haiqin;GUO Kangyue;HE Jiajun;CAO Zhiyun;ZHANG Youquan;LIN Minghe;CHEN Xuzheng(Academy of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China;Fujian Key Laboratory of Integrative Medicine on Geriatrics,Fuzhou,Fujian 350122,China;College of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China;College of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China;The Second Affiliated People's Hospital of Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350003,China)

机构地区：[1]福建中医药大学中西医结合研究院,福建福州350122 [2]福建省中西医结合老年性疾病重点实验室,福建福州350122 [3]福建中医药大学中西医结合学院,福建福州350122 [4]福建中医药大学药学院,福建福州350122 [5]福建中医药大学附属第二人民医院,福建福州350003

出　　处：《福建中医药》2023年第8期23-26,30,共5页Fujian Journal of Traditional Chinese Medicine

基　　金：福建省自然科学基金项目(2019J01479,2020J01747,2021J01939)。

摘　　要：目的 探讨芪灵扶正清解方(QFQ)对Huh-7细胞能量代谢及糖酵解相关蛋白的影响。方法CCK8法检测0、62.5、125、250、500μg/mL QFQ醇提物干预Huh-7细胞24、48和72 h后的细胞活力。将Huh-7细胞分为0、62.5、125、250、500μg/mL组,分别予相应浓度的QFQ醇提物干预24 h,应用细胞能量代谢分析仪检测线粒体和糖酵解ATP产生速率、实时耗氧率、胞外酸化率、糖酵解速率和质子流出速率;Western blot检测Huh-7细胞缺氧诱导因子1α(HIF-1α)、己糖激酶(HK)、葡萄糖转运蛋白(Glut)1以及Glut3的蛋白表达量。结果(1)与0μg/mL组比较,125、250、500μg/mL QFQ醇提物干预Huh-7细胞24、48和72 h后细胞活力明显降低(P<0.01)。(2)与0μg/mL组比较,62.5、125、250μg/mL组Huh-7细胞线粒体和糖酵解ATP产生速率比值明显提高(P<0.05),125、250μg/mL组Huh-7细胞糖酵解速率明显降低(P<0.05或P<0.01);与0μg/mL组比较,随着检测时间延长,62.5μg/mL组Huh-7细胞实时耗氧率呈升高趋势(P<0.05),250μg/mL组Huh-7细胞胞外酸化率和质子流出速率呈降低趋势(P<0.05)。(3)与0μg/mL组比较,62.5、125、250、500μg/mL组Huh-7细胞的HIF-1α、HK、Glut1、Glut3蛋白表达量均明显降低(P<0.01或P<0.05)。结论 QFQ通过调控细胞能量代谢和糖酵解相关蛋白HIF-1α、HK、Glut1和Glut3的表达,提高线粒体能力,抑制糖酵解能力,促进有氧呼吸,从而有效抑制Huh-7细胞的增殖。Objective:To explore the effect of Qiling Fuzheng Qingjie formula(QFQ)on energy metabolism and glycolysis related proteins in Huh-7 Cells.Methods:CCK8 method was used to detect the cell viability of Huh-7 cells after intervention with 0,62.5,125,250,and 500μg/mL QFQ ethanol extract for 24,48,and 72 hours.The Huh-7 cells were divided into 0,62.5,125,250,and 500μg/mL group,and were respectively treated with corresponding concentrations of QFQ ethanol extract for 24 hours.The cell energy metabolism analyzer was used to detect mitochondrial and glycolytic ATP production rates,real-time oxygen consumption rate,extracellular acidification rate,glycolysis rate,and proton outflow rate.Western blot was used to detect the protein expression of hypoxia inducible factor-1α(HIF-1α),hexokinase(HK),glucose transporter(Glut)1,and Glut3 in Huh-7 cells.Results:(1)Compared with the 0μg/mL group,the cell viability of Huh-7 cells significantly decreased after 24,48,and 72 hours of intervention with 62.5,125,250,and 500μg/mL QFQ ethanol extract.(2)Compared with the 0μg/mL group,the ratio of mitochondrial and glycolytic ATP production rate of Huh-7 cells in the 62.5,125,and 250μg/mL groups significantly increased(P<0.05),while the glycolytic rate of Huh-7 cells in the 125 and 250μg/mL groups significantly decreased(P<0.05 or P<0.01).Compared with the 0μg/mL group,as the detection time prolonged,the real-time oxygen consumption rate of Huh-7 cells in the 62.5μg/mL group showed an increasing trend(P<0.05),while the extracellular acidification rate and proton outflow rate of Huh-7 cells in the 250μg/mL group showed a decreasing trend(P<0.05).(3)Compared with 0μg/mL group,the proteins expression of HIF-1α,HK,Glut1 and Glut3 of Huh-7 cells in the 62.5,125,250 and 500μg/mL groups significantly decreased(P<0.05 or P<0.01).Conclusion:QFQ effectively inhibits the proliferation of Huh-7 cells by regulating cell energy metabolism and glycolysis related proteins expression of HIF-1α,HK,Glut1,and Glut3,improving mitochondrial abilit

关 键 词：肝癌 芪灵扶正清解方 HUH-7细胞 ATP 糖酵解 

分 类 号：R285[医药卫生—中药学]