氯贝丁酯影响肝线粒体功能而致肝损伤的观察

目的 研究氯贝丁酯对离体小鼠肝线粒体和体外培养的AML-12肝细胞损伤的作用机制。方法 用荧光染料JC-1对线粒体膜电位进行标记，检测膜电位变化。通过DCFDA的标记，检测对线粒体膜自由基的影响。加入环孢素（CsA）和抗氧化剂维生素C、甲磺酸去铁胺、过氧化氢酶等，观察对细胞的保护作用。结果 当氯贝丁酯浓度大于0.3mmol/L时，能迅速降低肝细胞中的线粒体以及小鼠肝脏离体线粒体的膜电位。CsA通过拮抗线粒体膜通透性，抑制膜电位下降，从而抑制细胞死亡。用氯贝丁酯处理过的肝细胞及线粒体内自由基显著增加。抗氧化剂维生素C、甲磺酸去铁胺、过氧化氢酶等能够保护细胞，抑制安妥明引起的损伤和死亡。结论 安妥明通过影响线粒体功能，从而诱发过多的自由基产生，最终导致细胞死亡。

Clofibrate induced hepatotoxicity by mitochondrial damage

Objective To explore the effect of clofibrate on freshly isolated mouse liver mitochondria and a cultured hepatocyte cell line, AML-12. Methods  Mitochondrial membrane potential was determined by using the fluorescent dye, JC-1. Levels of reactive oxygen species were measured with the fluorescent probes DCFDA. CsA and antioxidants such as Vitamin C, deferoxamine and catalase were used to protect cells. Results Application of clofibrate at concentrations greater than 0.3mmol/L rapidly collapsed the ΔΨm both in liver cells and in isolated mitochondria. The loss of ΔΨm occurred prior to cell death, as revealed by the protective effect of cyclosporin A (CsA) on the decrease in ΔΨm. Treatment  of hepatocytes with clofibrate caused a significant increase in intracellular and mitochondrial ROS. Antioxidants such as Vitamin C, deferoxamine, and catalase were able to protect cells against lofibrate-induced loss of viability. Conclusion  Clofibrate may impair mitochondrial function, stimulates formation of ROS, and eventually contributes to cell death.