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观察虾青素(astaxanthin)对呼吸链复合体Ⅳ抑制剂叠氮钠(NaN3)损伤的人胎肝L-02细胞保护作用,并初步探讨其作用机制。100 mmol·L-1 NaN3用于构建肝损伤细胞模型,通过测定不同浓度虾青素(0.01、0.10、1.00及10.00 nmol·L-1)对损伤细胞存活率(MTT检测)、细胞内活性氧(reactive oxygen species,ROS)水平(DCFH-DA检测)、细胞凋亡率(Annexin V-FITC/PI双染法)以及线粒体膜电位(mitochondrial membrane potential,MMP)水平(JC-1法)的影响,发现虾青素能抑制损伤细胞晚期凋亡;对细胞存活率和MMP的保护作用呈现先增加后降低的非剂量依赖性关系,其中0.10 nmol·L-1虾青素表现为较强的保护作用;实验浓度范围内的虾青素并不能显著降低细胞内ROS水平(P>0.05)。为进一步探讨虾青素对损伤细胞的保护作用,人工制备平面双层磷脂膜(planar bilayer lipid membrane,BLM)模拟线粒体膜,测定不同浓度虾青素(0.1%、2.0%、10.0%)对H+的传递能力。结果显示,虾青素对H+传递效率无剂量依赖性,中等浓度(2.0%)的虾青素能够较高效率地传递H+。结果提示,虾青素对NaN3损伤的人胎肝细胞的保护作用与其直接淬灭ROS的抗氧化功能无关,而可能是通过适当浓度下的虾青素对H+的高效传递进而维持线粒体膜电位实现的。
To observe the protective effect of astaxanthin on human fetal liver L-02 cells damaged by sodium azide (NaN3), a respiratory chain complex inhibitor, and to explore its mechanism. 100 mmol·L-1 NaN3 was used to establish a model of hepatic injury. The effects of different concentrations of astaxanthin (0.01,0.10,1.00 and 10.00 nmol·L-1) on the injury cell survival rate (MTT assay), intracellular reactive oxygen species (DCFH-DA assay), Annexin V-FITC / PI double staining and mitochondrial membrane potential (JC-1) , Astaxanthin was found to inhibit the late apoptosis of injured cells; the cell survival rate and the protective effect of MMP showed an increased and then decreased non-dose-dependent relationship, of which 0.10 nmol·L-1 astaxanthin showed strong protection Astaxanthin in the experimental concentration range did not significantly reduce intracellular ROS levels (P> 0.05). To further explore the protective effect of astaxanthin on injured cells, a planar bilayer lipid membrane (BLM) was prepared to simulate mitochondrial membrane. The effects of different concentrations of astaxanthin (0.1%, 2.0%, 10.0% The ability to pass. The results showed that astaxanthin had no dose-dependent effect on the transfer efficiency of H +, while medium concentration (2.0%) of astaxanthin delivered H + efficiently. The results suggest that the protective effect of astaxanthin on NaN3-injured human fetal hepatocytes is not related to the direct anti-oxidative function of ROS, but may be achieved by the efficient transfer of astaxanthin at an appropriate concentration to maintain mitochondrial membrane potential of.