大黄含药血清对乳鼠原代肝细胞能量代谢的影响

摘要

目的 探讨大黄含药血清对乳鼠原代肝细胞能量代谢的影响.方法 制备大黄含药血清;以胶原酶消化法获取乳鼠原代肝细胞,将细胞随机分为空白血清组、含药血清组,分别用20％空白血清培养液、20％大黄含药血清培养液干预24 h.采用生化法检测细胞内三磷酸腺苷(ATP)酶活性,高效液相色谱法检测细胞内ATP含量及细胞能荷,罗丹明123荧光染色法检测细胞线粒体膜电位.结果 与空白血清组比较,含药血清组Na+-K+-ATP酶、Ca2+-Mg2+-ATP酶活性低(P均＜0.05),ATP含量低(P＜0.05),两组能荷比较差异无统计学意义(P＞0.05).与空白血清组比较,含药血清组细胞线粒体膜电位低(P＜0.05).结论 大黄含药血清可能通过调整乳鼠原代肝细胞线粒体功能来抑制其能量代谢,降低其能量储备.%Objective To observe the effect of serum containing Dahuang (rhubarb) on energy metabolism of rat primary hepatocytes.Methods Medicated serum was prepared on SD rats.Hepatocytes were isolated from SD neonatal rats by collagenase digestion and then were randomly divided into the blank serum group and medicated serum group.Cells in the two groups were cultured with 20％ blank serum and 20％ serum containing Dahuang (rhubarb) for 24 hours,respectively.The activities of Na+-K +-ATPase,and Ca2+-Mg2+-ATPase were detected with spectrophotometer.The content of ATP,ADP,and AMP was tested with high performance liquid chromatography (HPLC) as well as energy charge.The mitochondrial membrane potential was analyzed with rhodamine 123 by laser confocal microscope.Results Compared with the blank serum group,the activities of Na +-K+-ATPase and Ca2+-Mg2+-ATPase decreased (all P ＜ 0.05) and the content of ATP decreased in the medicated serum group (P ＜ 0.05).There was no significant difference in the level of energy charge between the two groups (P ＞ 0.05).Compared with the blank serum group,the mitochondrial membrane potential decreased in the medicated serum group (P ＜ 0.05).Conclusion The serum containing Dahuang (rhubarb) may inhibit the energy metabolism and reduce the energy reserve of rat primary hepatocytes by regulating mitochondrial function.