甘草次酸和甘草酸对离体大鼠肝细胞膜效应的电镜观察

目的研究甘草次酸和甘草酸对离体大鼠肝细胞的膜效应及膜结合位点。方法应用体循环灌注胶原酶法分离正常大鼠肝细胞，在肝细胞培养中分别加入D-Hanks液、猪苓多糖、甘草酸及甘草次酸，24h后收集细胞。分别作扫描电镜和透射电镜观察膜效应，应用胶体金探针法检测正常肝细胞表面甘草次酸及甘草酸的结合位点。结果扫描电镜示加入甘草次酸的肝细胞表面微绒毛缺乏，呈现大量大小不等的光滑的囊泡状隆起；胶体金探针实验示透射电镜下可见加入甘草次酸?牛血清白蛋白-胶体金颗粒探针的正常肝细胞表面覆有密集的黑色胶体金颗粒。结论甘草次酸和甘草酸对大鼠离体肝细胞具有明显的生物膜效应；肝细胞膜上有与甘草次酸、甘草酸相结合的位点；且以甘草次酸更为显著。

Effects and Binding Sites of Glycyrrhetinic Acid and Glycyrrhizin on Isolated Rat Hepatocyte Membrane

Purpose To investigate the effects and binding sites of glycyrrhetinic acid and glycyrrhizin on isolated rat hepatocyte membrane. Methods Hepatocytes were isolated from the normal rat livers by collagenase pouring into circulation and were respectively cultured with D?Hanks solution,Zhuling Duotang,glycyrrhizin and glycyrrhetinic acid in vitro .After 24 hours,The hepatocytes were collected and observed under electron microscope and used colloidal gold probe to respectively detect the binding sites of the glycyrrhizin and glycyrrhetinic acid on the surface of the hepatocytes. Results Under scanning electron microscope,heptocytes treated with the glycyrrhetinic acid exhibited a surface with abundant smooth surfaced bleb like processes of various size and were devoid of microvilli;Under transmission electron microscope,concentrated colloidal gold granules in black were obviously seen at the surface of hepatocytes treated with glycyrrhetinic acid bovine serum albumin colloidal gold granule probe. Conclusions The biological effect and the binding sites of glycyrrhetinic acid on the surface of hepatocytes were more remarkable than glycyrrhizin.