积雪草酸对大鼠离体肝细胞线粒体损伤的防护作用  

Protection of Asiatic Acid on Damage in Rat Hepatic Mitochondria in Vitro

作　　者：赵华龙[1] 毛涵[1,2] 刘佳[3] 薛菲[1] 汤新慧[1] ZHAO Hua-iong;MAO Han;LIU Jia;XUE Fei;TANG Xin-hui(Key Laboratory of Coastal Wetland Bioresourees and Environmental Protection of Jiangsu Province,Yancheng Teachers University,Yancheng 224051,China;College of Biotechnology and Pharmaceutical Engineering,Nanjing University of Technology,Nanjing 210009,China;School of Pharmacy,Jiangsu University,Zhenjiang 212013,China)

机构地区：[1]盐城师范学院江苏省滩涂生物资源与环境保护重点建设实验室,江苏盐城224051 [2]南京工业大学生物与制药工程学院,江苏南京210009 [3]江苏大学药学院,江苏镇江212013

出　　处：《食品工业科技》2018年第16期286-289,295,共5页Science and Technology of Food Industry

基　　金：江苏省"青蓝工程"科技创新团队项目;江苏省滩涂生物资源与环境保护重点建设实验室项目(JLCBE10006)

摘　　要：为考察积雪草酸对大鼠肝细胞线粒体镉损伤的防护作用及作用机制,建立Cd^(2+)诱发的大鼠离体肝细胞线粒体高通透性转运(MPT)模型,将线粒体分为对照、积雪草酸(AA)低、中、高剂量(25、50、100μmol/L)、钌红(RR)5组,用分光光度法测定线粒体肿胀度,用荧光探针罗丹明(Rh123)检测线粒体膜电位,用荧光染料fura-2/AM测定线粒体游离钙释放,Western blot法检测线粒体内细胞色素c(Cyt c)和凋亡诱导因子(AIF)的释放。结果表明,正常大鼠肝细胞线粒体中加入10μmol/L Cd^(2+)即可引起明显的线粒体肿胀、膜电位耗散以及游离钙的释放,并诱发线粒体内Cyt c和AIF的释放,而25、50、100μmol/L AA预处理能明显抑制上述过程,100μmol/L AA对Cd^(2+)诱导的线粒体肿胀、膜电位耗散、游离钙释放以及线粒体Cyt c和AIF的释放的抑制率分别为:49.5%、41.9%、47.9%、62.2%和66.3%,表明AA能通过抑制肝细胞线粒体MPT过程,对抗Cd^(2+)引起的线粒体损伤,发挥其肝细胞保护作用。To investigate the protective effect and mechanism of asiatic acid on mitochondrial cadmium injury in rat liver cells,Cd^2＋ induced hepatic mitochondrial permeability transition（MPT）was established,and the mitochondria were randomly divided into five groups of control,25,50,100 mol/L AA and RR（Ruthenium Red）group.Mitochondrial swelling was assessed by measuring the absorbance of their suspension,mitochondrial membrane potential was evaluated according to the uptake of the fluorescent dye rhodamine 123（Rh123）,the intramitochondrial Ca^2＋ level was assayed by Ca^2＋ indicator dye fura-2/AM,and transfer of cytochrome c（Cyt c）and apoptosis-inducing factor（AIF）from the intermembrane space to the cytoplasm was determined by Western blot analysis.The results showed that obvious mitochondrial swelling,loss of mitochondrial membrane potential,and release of matrix Ca^2＋occurred after the addition of 10μmol/L Cd^2＋.In addition,Cyt c and AIF releasing were detected in the mitochondrial supernatant.However,preincubation with 20,50,100μmol/L AA could significantly block the above changes.Inhibition rates of 100μmol/L AA on mitochondrial swelling,mitochondrial membrane potential dissipating,and mitochondrial Ca^2＋,Cyt c and AIF releasing were 49.5%,41.9%,47.9%,62.2%and 66.3%,respectively.The results suggested that AA had significant protection from Cd^2＋-induced rat hepatic mitochondria damage and the mechanisms might relate to its direct inhibitory effect on hepatic MPT.

关 键 词：积雪草酸 肝细胞线粒体 线粒体高通透性转运 细胞色素C 凋亡诱导因子 

分 类 号：TS201.4[轻工技术与工程—食品科学]