单 位：
南京工业大学生物与制药工程学院;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;江苏大学药学院
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
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。
译 名：
Protection of Asiatic Acid on Damage in Rat Hepatic Mitochondria in Vitro
作 者：
ZHAO Hua-long;MAO Han;LIU Jia;XUE Fei;TANG Xin-hui;Key Laboratory of Coastal Wetland Bioresources and Environmental Protection of Jiangsu Province,Yancheng Teachers University;College of Biotechnology and Pharmaceutical Engineering,Nanjing University of Technology;School of Pharmacy,Jiangsu University;
单 位：
ZHAO Hua-long%MAO Han%LIU Jia%XUE Fei%TANG Xin-hui%Key Laboratory of Coastal Wetland Bioresources and Environmental Protection of Jiangsu Province,Yancheng Teachers University%College of Biotechnology and Pharmaceutical Engineering,Nanjing University of Technology%School of Pharmacy,Jiangsu University
关键词：
asiatic acid(AA);;hepatic mitochondria;;mitochondrial permeability transition(MPT);;cytochrome c(Cytc);;apoptosis inducing factor(AIF)
摘 要：
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.
