吡啶甲酸铬对新生仔猪原代肝细胞氧化作用的影响

The Effect of Chromium Picolinate on Oxidative Damage in New Born Porcine Hepatocyte

【目的】研究不同剂量吡啶甲酸铬(CrPic)对新生仔猪原代肝细胞氧化作用的影响。【方法】选用3日龄新生仔猪作为细胞供体,用剪切消化法分离肝细胞,分别用0、8、200、400μmol·L-1的吡啶甲酸铬处理细胞48h,研究吡啶甲酸铬对细胞内活性氧(ROS)水平、丙二醛(MDA)的含量、培养上清液中乳酸脱氢酶(LDH)活性以及对细胞DNA单链断裂(彗星试验)的影响。【结果】与对照组相比,各处理组ROS水平、LDH活力和彗星试验指标无显著差异(P>0.05),8μmol·L-1组MDA含量显著降低(P<0.05);与8μmol·L-1组相比,400μmol·L-1处理组ROS、MDA、LDH和DNA损伤程度均显著升高(P<0.05),随着吡啶甲酸铬添加水平的升高,ROS、LDH分别呈先下降后上升的二次曲线型变化趋势(P<0.05)。【结论】体外条件下适量的吡啶甲酸铬能抑制肝细胞内脂质过氧化物的生成,增强猪肝细胞的抗氧化能力;400μmol·L-1吡啶甲酸铬对猪肝细胞无抗氧化作用,也不会引起肝细胞的氧化损伤。

[ Objective ] This experiment was designed to assess the effect of different levels of chromium picolinate （CrPic） on oxidation in new-born porcine hepatocyte. [Method] Three-day-old new born piglets were chosen as hepatocyte donors, hepatocytes were isolated enzymatic and mechanical methods. Porcine hepatocytes were cultured in medium in which added 0, 8, 200, 400 μmol·L^-1 of CrPic for 48 hours. The levels of reactive oxygen species （ROS）, the contents of malondialdehyde （MDA） in hepatocytes, the activities of lactate dehydrogenase （LDH） in medium and DNA strand breaks （by comet assay） were examined to investigate the effect of CrPic on porcine hepatocytes. [ Result ] Compared with control, the treatment groups have no significant differences in ROS level in hepatocytes, LDH activities in medium and DNA strand breaks （P〉0.05）, however, 8 pmol.L-1 CrPic was significantly decreased MDA content in hepatocytes （P〈0.05）. Compared with 8 μmol·L^-1 CrPic group, the other parameters were markedly increased in 400μmol·L^-1 CrPic group （P〈0.05）. Meanwhile, with the levels of CrPic increasing, analysis revealed a significant quadratic trend between CrPic and ROS （or LDH） （P〈0.05）. [Conclusion] The appropriate amount of CrPic inhibits lipid peroxidation in cells, enhances antioxidation of porcine hepatocyte, however, 400 μmol·L^-1 of CrPic has no effects on the antioxidation and oxidative damage.