槲皮素-3-O-β-D-葡萄糖醛酸苷对游离脂肪酸诱导HepG2细胞脂肪变性的作用  

Effect of quercetin-3-O-β-D-glucuronide on free fatty acid induced steatosis in HepG2 cells

作　　者：王路路[1] 张志超[1] 吴诉诉 尚靖[1] 

机构地区：[1]中国药科大学新药筛选中心,南京210009

出　　处：《中国药科大学学报》2015年第5期587-593,共7页Journal of China Pharmaceutical University

基　　金："十二五"国家科技支撑计划资助项目(No.2012BAI30B01)~~

摘　　要：探讨槲皮素-3-O-β-D-葡萄糖醛酸苷(quercetin-3-O-β-D-glucuronide,Q3GA)对游离脂肪酸诱导的人源肝癌细胞HepG2细胞脂质蓄积的甘油三酯调节和氧化应激的作用及其可能的相关机制。采用油红染色检测Q3GA对游离脂肪酸诱导的HepG2细胞中脂滴含量的影响,并同时检测其对甘油三酯和胆固醇的作用。DCFH-DA法检测Q3GA对HepG2细胞脂质蓄积引起的活性氧(ROS)的变化;硫代巴比妥酸法和黄嘌呤氧化酶法分别测定丙二醛(MDA)的含量和超氧化物歧化酶(SOD)的活性。RT-PCR分析脂肪酸氧化相关的基因过氧化物酶体增殖物受体(PPARα)、肉毒碱棕榈酰转移酶(CPT1A)、中链酰基辅酶A脱氢酶(MCAD)、细胞色素P450 4A11(CYP4A11)、乙酰辅酶A氧化酶(ACO)的表达情况。实验结果显示,Q3GA可剂量依赖性降低FFA诱导的Hep G2细胞脂质蓄积和甘油三酯的含量,但未降低胆固醇的含量。同时可改善脂肪酸氧化引起ROS,MDA的升高以及SOD的降低。另外,Q3GA在一定浓度下可上调脂肪酸β氧化相关基因PPARα、CPT1A、MCAD的表达,而对CYP4A11和ACO的表达没有促进作用。综上所述,Q3GA可抵抗脂肪酸氧化引发肝细胞的氧化应激损伤,保护HepG2细胞,降低游离脂肪酸诱导HepG2细胞脂质蓄积和甘油三酯的含量,其调节机制可能与其对HepG2细胞中游离脂肪酸氧化有关。The effects of quercetin-3-O-β-D-glucuronide （Q3GA）on the triglyceride metabolism and oxidative stress in steatotic HepG2 cells and the underlying mechanism were investigated in this study.Significant fat accu-mulation was documented by Oil Red O staining;intracellular triglyceride levels were detected by triglyceride（TG）enzymatic assay.DCFH-DA staining assay was performed to observe reactive oxygen species （ROS）pro-duction of HepG2 cells.The level of malondialdehyde（MDA）and superoxide dismutase（SOD）were assayed by thibabituric acid method and xanthine oxidase method.Changes in the mRNA expression of peroxisome prolifera-tor-activated receptorα（PPARα）;carnitine palmitoyltransferase 1 A（CPT1 A）;medium chain acyl-CoA dehydro-genase （MCAD）;cytochrome P450 4A11（CYP4A11）and acyl-CoA oxidase（ACO）;which are related with fatty acid oxidation were assessed by RT-PCR.Our results showed that Q3GA obviously reduced fat deposition and TG content.At the same time;Q3 GA decreased MDA content and significantly increased the SOD activity with reduced ROS production.Moreover;the PPARα;CPT1A;MCAD expression-related fatty acid βoxidation was upregulated with the treament of Q3GA;while without any change of the expression of CYP4A11;ACO.In conclu-sion;Q3GA prevents FFA-induced HepG2 cell steatosis;and enhances mitochondrial fatty acidβoxidation;which＆amp;nbsp;may partly be related to its anti-oxidation ability.

关 键 词：槲皮素-3-O-β-D-葡萄糖醛酸苷 HEPG2细胞 脂质沉积 氧化应激 

分 类 号：R575.5[医药卫生—消化系统]