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.
作者
赵华龙
毛涵
刘佳
薛菲
汤新慧
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)
出处
《食品工业科技》
CAS
CSCD
北大核心
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)
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
asiatic acid （ AA ）
hepatic mitochondria
mitochondrial permeability transition （ MPT ）
cytochrome c （ Cyt c ）
apoptosis inducing factor（AIF）
