目的:研究三萜类化合物对半乳糖胺(D-GalN)和四氯化碳(CCl4)损伤原代培养大鼠肝细胞的保护作用及其机制。方法:两步灌流法分离大鼠肝细胞进行原代培养,评价积雪草酸(asiaticacid,AA)和β-甘草次酸(β-glycyrrhetinicacid,GA)对D-GalN和CCl4损伤原代培养肝细胞的保护作用。光镜评价细胞生长形态,MTT法测定细胞活性,测定细胞上清天冬氨酸氨基转移酶(AST)和乳酸脱氢酶(LDH);并以荧光分光光度法测定细胞中活性氧(ROS),细胞上清活性氮终产物(NOx)和细胞内谷胱甘肽(GSH)含量;用JC-1法测定细胞线粒体膜电位(ΔΨm)。结果:AA和GA均可显著抑制D-GalN所致的AST和LDH升高(P<0.05),AA尚能提高细胞存活率(P<0.05);AA和GA也能显著抑制CCl4所致的LDH释放(P<0.05)。AA和GA均显著减少两种化学损伤细胞ROS生成和NOx释放,明显改善D-GalN所致细胞线粒体ΔΨm的降低;AA尚显著抑制两种化学损伤细胞内GSH降低。结论:三萜类化合物对D-GalN和CCl4致原代培养大鼠肝细胞损伤有保护作用,其机制与抑制细胞ROS、NOx生成和GSH降低相关,对D-GalN损伤尚有改善线粒体膜电位作用。 Objective: To study the protective effect of triterpene on primary cultured rat hepatocytes induced by galactosamine (D-GalN) and carbon tetrachloride (CCl4) and its mechanism. Methods: Two-step perfusion method was used to isolate rat hepatocytes for primary culture. The effects of asiatic acid (AA) and β-glycyrrhetinic acid (GA) on D-GalN and CCl4 injury in primary cultured hepatocytes The protective effect. The cell growth morphology was evaluated by light microscopy. The activity of aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) were measured by MTT assay. The levels of reactive oxygen species (ROS) (NO) and intracellular glutathione (GSH) contents in the cell supernatant were measured. The mitochondrial membrane potential (ΔΨm) was measured by JC-1 method. Results: Both AA and GA could significantly inhibit the increase of AST and LDH induced by D-GalN (P <0.05), but AA could still increase the cell survival rate (P <0.05). AA and GA also significantly inhibited the increase of CCl4 LDH release (P <0.05). Both AA and GA significantly reduced ROS production and NO x release from both chemically damaged cells and significantly decreased the ΔΨm of mitochondria induced by D-GalN. AA also significantly inhibited the reduction of GSH in both chemically damaged cells. CONCLUSION: Triterpenoids have a protective effect on primary cultured rat hepatocytes induced by D-GalN and CCl4, and its mechanism is related to the inhibition of ROS and NOx production and GSH reduction. D-GalN injury may improve mitochondrial membrane potential effect.