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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 ,2 4h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
Objective To study the membrane effect and membrane-binding site of glycyrrhetinic acid and glycyrrhizic acid on isolated rat hepatocytes. Methods Normal rat hepatocytes were isolated by systemic perfusion collagenase method. D Hanks solution, Polyporus umbellatus polysaccharide, glycyrrhizic acid and glycyrrhetinic acid were added to the hepatocyte culture. Cells were harvested 24 hours later. The membrane effect was observed by scanning electron microscopy and transmission electron microscopy respectively. The binding sites of glycyrrhetinic acid and glycyrrhizic acid on the surface of normal liver cells were detected by colloidal gold probe method. RESULTS: Scanning electron microscopy showed that glycyrrhetinic acid-added hepatocytes lacked surface microvilli and showed a large number of smooth vesicular bulges of varying sizes. Colloidal gold probe experiments showed that glycyrrhetinic acid bovine serum albumin colloid was added under transmission electron microscope. Gold particle probes are covered with dense black colloidal gold particles on the surface of normal liver cells. Conclusion Glycyrrhetinic acid and glycyrrhizic acid have obvious biofilm effect on rat hepatocytes in vitro. The sites of hepatic cell membrane are combined with glycyrrhetinic acid and glycyrrhizic acid; and glycyrrhetinic acid is more significant.