解偶联蛋白2在槲皮素改善肝细胞脂肪变性中的作用研究  

Study of the role of uncoupling protein 2 in quercetin improving hepatocyte steatosis

作　　者：李丽红 李欣 李硕 邰杨浩 梁力中 高颖[1] 刘玉 荀阳烨 郭建红[1] LI Li-hong;LI Xin;LI Shuo;TAI Yang-hao;LIANG Li-zhong;GAO Ying;LIU Yu;XUN Yang-ye;GUO Jian-hong(School of Basic Medical Sciences,Shanxi Medical University,Taiyuan 030001,Shanxi Province,China;Second Hospital of Shanxi Medical University,Shanxi Medical University,Taiyuan 030001,Shanxi Province,China;First Hospital of Shanxi Medical University,Shanxi Medical University,Taiyuan 030001,Shanxi Province,China;Department of Medicine,Linfen Vocational and Technical College,Linfen 041000,Shanxi Province,China)

机构地区：[1]山西医科大学基础医学院,山西太原030001 [2]山西医科大学第二临床医学院,山西太原030001 [3]山西医科大学第一临床医学院,山西太原030001 [4]临汾职业技术学院医学系,山西临汾041000

出　　处：《中国临床药理学杂志》2022年第16期1892-1896,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的探讨在槲皮素通过减少活性氧(ROS)改善游离脂肪酸(FFA)诱导的肝细胞脂肪变性中解偶联蛋白2(UCP2)的作用。方法用FFA诱导肝细胞建立肝脂肪变性体外模型。将HepG2细胞分为正常组、模型组和槲皮素组。正常组用正常培养基培养;模型组用1 mmol·L^(-1)混合FFA溶液(油酸:棕榈酸=2:1)干预24 h;槲皮素组用1 mmol·L^(-1)FFA混合液先干预24 h,后用30μmol·L^(-1)槲皮素处理24 h。以GPO-PAP法测定三酰甘油(TG)含量;以酶联免疫吸附试验(ELISA)检测肿瘤坏死因子-α(TNF-α)蛋白水平;以实时荧光定量PCR(RT-PCR)法检测肉碱棕榈酰转移酶Iα(CPT1α)和UCP2 mRNA表达水平。结果正常组、模型组和槲皮素组的TG含量分别为(0.39±0.03),(2.17±0.31)和(1.12±0.16)mmol·gprot^(-1);这3组的TNF-α含量分别为(0.33±0.01),(0.42±0.02)和(0.33±0.02)μg·L^(-1)。槲皮素组与模型组比较,TG和TNF-α含量明显减少,差异均有统计学意义(均P<0.01)。正常组、模型组和槲皮素组的CPT1αmRNA表达水平分别为1.00±0.23,0.27±0.02和1.79±0.17;这3组的UCP2 mRNA表达水平分别为1.00±0.13,0.51±0.05和1.84±0.10。槲皮素组与模型组比较,CPT1α和UCP2基因表达水平均明显上调,差异均有统计学意义(P<0.01,P<0.001),且UCP2和CPT1α之间呈显著正相关(P<0.01)。结论槲皮素可能通过诱导UCP2表达上调,抑制线粒体过量ROS和炎性因子TNF-α生成,从而增强线粒体脂肪酸β氧化功能,最终改善肝细胞脂肪变性。Objective To explore the role of uncoupling protein 2(UCP2)while quercetin alleviates free fatty acid(FFA)-induced hepatocyte steatosis through reduction of reactive oxygen species(ROS).Methods HepG2 cells were induced by FFA to establish the vitro model of hepatic steatosis.The three groups were divided into normal group(cultured with normal medium),model group[cultured with 1mmol·L^(-1)of the FFA mixture(oleate/palmitate,2:1 ratio)for 24 h]and quercetin group(treated with 1 mmol·L^(-1) of the FFA mixture for24 h first,and then treated with 30μmol·L^(-1) quercetin for 24 h).Triglyceride(TG)was determined with glycerol phosphate oxidase peroxidase method(GPO-PAP),tumor necrosis factor alpha(TNF-α)protein level was measured by enzyme-linked immunosorbent assay(ELISA);carnitine palmitoyl transferase 1α(CPT1α)and UCP2 mRNA were measured by real-time fluorescence quantitative PCR(RT-PCR).Results The TG of normal group,model group and quercetin group were(0.39±0.03),(2.17±0.31)and(1.12±0.16)mmol·gprot^(-1),respectively;the TNF-α protein in these three groups were(0.33±0.01),(0.42±0.02)and(0.33±0.02)μg·L^(-1),respectively.The TG and TNF-α of quercetin group were significantly decreased compared with that in model group(all P<0.05).The expression level of CPT1αmRNA in normal,model and quercetin groups were 1.00±0.23,0.27±0.02 and 1.79±0.17,respectively;the expression level of UCP2 mRNA in these three groups were 1.00±0.13,0.51±0.05 and 1.84±0.10,respectively.The expression levels of CPT1αand UCP2 were both significantly up-regulated of quercetin group compared with that of model group(P<0.01,P<0.001),and there was a significant positive correlation between UCP2 and CPT1α.Conclusion Quercetin enhances mitochondrial fatty acid β-oxidation function and ultimately improves hepatocyte steatosis,which may be achieved by inducing up-regulation of UCP2 expression and inhibiting the production of excessive mitochondrial ROS and inflammatory factor TNF-α.

关 键 词：槲皮素 脂肪变性 解偶联蛋白2 活性氧 脂肪酸β氧化 

分 类 号：R96[医药卫生—药理学]