甘草甜素甘草多糖和光甘草定对小鼠巨噬细胞的毒性与免疫功能的调节

Effects of glycyrrhizin,glycyrrhiza polysaccharide and glabridin on the toxicity and immunologic function of murine peritoneal macrophages

为探讨甘草的3种主要成分甘草甜素、甘草多糖和光甘草定对小鼠腹腔巨噬细胞的毒性与免疫功能的调节作用,将3种药物均设6个质量浓度(0.1、1、5、20、100和400μg/mL),分别与巨噬细胞作用12、24和36h后,利用CCK-8细胞活性检测试剂盒检测药物对巨噬细胞的毒性作用;荧光显微镜观察无细胞毒性浓度下对巨噬细胞吞噬大肠杆菌的影响,用ELISA试剂盒检测3种药物对巨噬细胞分泌细胞因子的影响。结果显示,400μg/mL甘草甜素和光甘草定对细胞有一定毒性;所有试验浓度的甘草甜素和甘草多糖均能显著增强巨噬细胞的吞噬能力(P<0.01),而仅5、20和100μg/mL光甘草定有这种作用(P<0.01);所有试验浓度的3种药物均能显著增强巨噬细胞分泌IL-1β、IL-6、IL-12和TNF-α的能力(P<0.01),高剂量抑制IL-4、IL-10和TGF-β的分泌(P<0.01),而低剂量没有明显变化。结果表明,甘草的3种主要成分甘草甜素、甘草多糖和光甘草定对小鼠腹腔巨噬细胞基本无毒性,呈剂量依赖性地增强巨噬细胞吞噬能力和分泌促炎性细胞因子以及降低抑炎性细胞因子的分泌。

The aim of this study was to evaluate the effects of glycyrrhizin,glycyrrhiza polysaccharide and glabridin on the toxicity and immunologic function of murine peritoneal macrophages. CCK-8 kit was used to test cytotoxicity after macrophages being co-cultured with drug concentration of 0.1,1,5,20,100 and 400μg/mL for 12,24 and 36 h. Phagocytosis of Escherichia coli labeled with FITC by macrophage was observed by fluorescence microscopy and evaluated using phagocytic percentage（PP） and phagocytic index （PI）. The levels of IL-lt？, IL-4, IL-6, IL-10, IL-12, TNF-α, and TGF-β were measured by ELISA. In result, only macrophage viability was reduced when being treated with 400 μg/mL of glycyrrhizin and glabridin. All concentrations drastically enhanced phagocytosis in glycyrrhizin and glycyrrhiza polysaccharide in a dose-dependent manner, except 5,20 and 100μg/mL of glabridin. A dramatic increase of the levels of IL-lfl, IL-6,IL-12 and TNF-a was found in the drug group compared with the control group. The 20,100 or 400μg/mL inhibited production of IL-4, IL-10 and TGF-β, but 0.1,1 or 5 μg/mL did not. The above results showed that glycyrrhizin,glycyrrhiza polysaccharide and glabridin could promote the phagocytosis of mac- rophage and regulate the immunologic functions.