莱菔硫烷对肝细胞的抗氧化能力和线粒体功能的影响  

Effect of sulforaphane on antioxidant capacity and mitochondrial function of hepatocytes in vitro and in rats with hepatic oxidative stress

作　　者：雷鹏 田思聪 滕春莹 孙瑶 单毓娟 李宝龙 LEI Peng;TIAN Sicong;TENG Chunying;SUN Yao;SHAN Yujuan;LI Baolong(Faculty of Food Science and Engineering,School of Chemistry and Chemical Engineering,Harbin Institute of Technology,Harbin,Heilongjiang Province,150001;Center of Drug Safety and Evaluation,Heilongjiang University of Chinese Medicine,Harbin,Heilongjiang Province,150040,China)

机构地区：[1]哈尔滨工业大学化工与化学学院食品科学与工程系,哈尔滨150001 [2]黑龙江省中医药大学药物安全评价中心,哈尔滨150040

出　　处：《第三军医大学学报》2020年第2期141-147,共7页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81573135)~~

摘　　要：目的探讨莱菔硫烷(sulforaphane,SFN)对肝细胞氧化应激和线粒体损伤的改善作用。方法建立高脂膳食诱导大鼠肝脏氧化应激体内模型和游离脂肪酸诱导HHL-5肝细胞氧化应激的体外模型。ELISA检测肝组织中丙二醛(malondialdehyde, MDA)的含量,Western blot测定大鼠肝脏组织中抗氧化酶血红素加氧酶-1(hemeoxygenase-1, HO-1),磷酸酰胺腺嘌呤二核苷酸醌氧化还原酶-1(NADH dehydrogenase quinone 1, NQO1)和谷胱甘肽-S-转移酶(glutathione S-transferase, GST)的蛋白表达。采用荧光显微镜观察HHL-5肝细胞的活性氧(reactive oxygen species, ROS)含量和线粒体膜电位;ELISA测定HHL-5肝细胞中谷胱甘肽(glutathione, GSH)和超氧化物歧化酶(superoxide dismutase, SOD)的水平。实时定量PCR法检测mtDNA含量。结果 SFN减少由高脂饲料引起MDA含量的增多(P<0.05),并上调大鼠肝组织中的抗氧化酶HO-1(P<0.05),NQO1(P<0.05)和GST(P<0.05)的蛋白表达。SFN降低由FFA提高HHL-5细胞的ROS水平(P<0.05),并提高其GSH(P<0.05)和SOD(P<0.05)的水平,以及线粒体膜电位(P<0.05)。此外,SFN显著提高HHL-5细胞的mtDNA含量(P<0.05)。结论 SFN通过增加肝细胞抗氧化能力改善高脂诱导的肝脏氧化应激损伤并对肝脏线粒体功能有保护作用。Objective To investigate the effect of sulforaphane(SFN) on oxidative stress and mitochondrial injury in hepatocytes. Methods We examined malondialdehyde(MDA) content by ELISA in the liver tissues and the protein expression of the antioxidant enzymes including hemeoxygenase-1(HO-1), NADH dehydrogenase quinone 1(NQO1) and glutathione S-transferase(GST) using Western blotting in the liver tissues from a rat model of hepatic oxidative stress established by high-fat diet. We also assessed the level of reactive oxygen species(ROS) and mitochondrial membrane protential by fluorescence microscopy, tested glutathione(GSH) and superoxide dismutase(SOD) levels using ELISA, and detected mitochondrial DNA(mtDNA) content using real-time quantitative PCR in cultured hepatocytes(HHL-5 cells) under fatty acid-induced oxidative stress. Results In the rat model of hepatic oxidative stress, SFN significantly reduced high-fat diet-induced elevation of MDA level(P<0.05) and up-regulated the protein expression of the antioxidant enzymes in the liver tissues(P<0.05). In cultured HHL-5 cells, SFN obviously reduced free fatty acid-induced elevation of ROS level(P<0.05) and increased GSH content(P<0.05), SOD level(P<0.05), mitochondrial membrane potential(P<0.05), and the content of mtDNA(P<0.05). Conclusion SFN can enhance the antioxidant capacity of hepatocytes to improve high-fat diet-induced hepatic oxidative stress in rats and provide protection of mitochondrial function in the hepatocytes.

关 键 词：莱菔硫烷 高脂膳食 线粒体功能 抗氧化能力 

分 类 号：R282.71[医药卫生—中药学] R322.47[医药卫生—中医学]