腰痛宁及其衍生方中几种皂苷有效部位的抗炎、免疫调节与骨细胞修复活性的比较

Comparison of Cellular Anti-inflammatory, Immunoregulatory and Chondrocyte-proliferative Activities of Several Saponins from Yaotongning Prescription and Its Derived Formulas

目的研究腰痛宁及其衍生方中几种皂苷有效部位的细胞抗炎、免疫调节与骨细胞修复活性,为风湿骨病处方候选药物筛选提供依据。方法在马钱子、麻黄生物碱与甘草等5种药材黄酮混合物的基础上,分别加入50%的甘草皂苷、三七皂苷、人参皂苷、川牛膝皂苷组成新的中药有效部位组方。采用半数有效浓度(EC50)或半数抑制浓度(IC50)评价各样品对巨噬细胞Ana-1分泌白介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)的促进作用;对脂多糖(LPS)诱导的Ana-1释放前列腺素(PG)E2的抑制作用;对IL-1β诱导的软骨细胞增殖的影响。同时采用最小二乘优化方法,计算各样品的EC50或IC50叠加值,根据EC50或IC50叠加值与实验值之间的差异分析各有效部位间的相互作用关系。结果 25%生物碱+25%黄酮+50%甘草皂苷组方的细胞抗炎、免疫和促进骨细胞修复活性优于其他组方,且该方中的各有效部位之间产生了较强的协同增效作用。结论以甘草皂苷为主的I号样品有良好的细胞抗炎、免疫调节及骨细胞修复活性,提示甘草皂苷可作为生物碱+黄酮+皂苷的风湿骨病处方中皂苷的优选有效部位。

Objective To compare the effects of different kinds of saponins from Yaotongning（YTN） and its derived formulas on cellular anti-inflammation,immunoregulation and chondrocyte proliferation. Methods Saponins of Radix Glycyrrhizae, Radix notoginseng, Radix Ginseng and Radix Cyathulae at 50 % were respectively added into the mixture consisting of alkaloids of Semen Strychni and Radix Ephedrae together with flavonoids extracted from five kinds of herbs such as Radix Glycyrrhizae. Half effective concentrations（EC50）or half inhibitory concentrations（IC50）were applied to evaluate the effects of the samples on promoting the secretion of interleukin 1β（IL-1β）,IL-6 and tumor necrosis factor-α（TNF-α） in macrophage（Ana-1）cells,on inhibiting the production of lipopolysaccharide（LPS）-induced PGE2 in Ana-1 cells, and on promoting the secretion of chondrocytes induced by IL-1β. Least square optimization method was applied to calculate the additive values of IC50/EC50 of each sample. The interactions among the active fractions in these samples were investigated by comparing the difference between the additive values and experimental values of IC50/EC50. Results The sample consisting of 25 % of alkaloids mixture,25 % of flavonoids mixture and 50 % of saponins of Radix Glycyrrhizae had better effect than other samples on cell anti-inflammation,immunoregulation and chondrocyte- proliferation. Moreover,strong synergistic effect among the effective fractions in this samples was observed. Conclusion Sample I mainly consisting of the saponins from Radix Glycyrrhizae works well at promoting chondrocytes proliferation,cellular anti-inflammation and immunoregulation. It is suggested the saponins from Radix Glycyrrhizae can be used as the preferred candidate effective fractions of saponins in the prescription which consists of alkaloids,flavonoids and saponins for curing rheumatoid bone diseases.