目的 探究以马钱子、麻黄生物碱为主要部位的7 种腰痛宁衍生方的细胞药理活性,并对细胞模型下有优效的组方进行体内药效学评价。方法 以50%马钱子生物碱+50%麻黄生物碱为组方基础,分别与腰痛宁胶囊组方药材及8 种常用风湿骨病组方中药的总黄酮、总皂苷、总挥发油/水提物及总多糖有效部位组合成7 个样品,同时以腰痛宁加黄酒药引为对照。测定各样品抑制小鼠巨噬细胞中前列腺素E₂(PGE₂)增殖的IC₅₀ 及促进小鼠巨噬细胞中白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)及肿瘤坏死因子(TNF-α)增殖的EC₅₀,并进行比较。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 实验值和叠加值之间的差异分析各有效部位间的相互作用关系。采用小鼠耳肿胀模型和小鼠皮肤迟发型超敏反应(DTH)模型进行最优组合的体内药效学评价。结果 25%马钱子生物碱+25%麻黄生物碱与总黄酮组合(4 号样品)具有最佳的综合细胞抗炎、免疫调节活性,且各模型下该组方中有效部位间有极强的协同作用;25%马钱子生物碱+25%麻黄生物碱分别与总皂苷、总挥发油/水提物及总多糖组合样品(5、6、7 号样品)的抗炎活性与4 号样品相当;5 和7 号样品有良好的综合免疫调节活性;25%马钱子生物碱+25%麻黄生物碱以及马钱子生物碱或麻黄生物碱与总黄酮组合样品(1、2、3 号样品)的药理活性均显著弱于4 号样品;小鼠体内药效学实验表明将4 号样品组合中25%马钱子生物碱+25%麻黄生物碱比例降为25%及5%时,具有较好的抗炎、免疫作用。结论 马钱子生物碱不宜单独与麻黄生物碱配伍,2 种生物碱不宜单独与总黄酮配伍,25%马钱子生物碱+25%麻黄生物碱与总黄酮、总皂苷、总挥发油/水提物及总多糖分别组合通常会产生协同或叠加作用,有助于增强25%马钱子生物碱+25%麻黄生物碱的细胞药理活性,25%马钱子生物碱+25%麻黄生物碱与总黄酮组合样品的细胞抗炎和免疫调节活性在小鼠体内亦得到体现,提示根据细胞实验结果筛选优化中药组方具有可行性。

Objective To investigate which kind of Chinese medicinal effective fractions will produce good comprehensive cellular pharmacological activities when they were combined with alkaloids from Strychnos nux-vomica and Ephedra sinica, and to evaluate in vivo pharmacological activities of the effective fraction combination screened by cellular experiments. Methods The equal proportion mixture of alkaloids from S. nux-vomica and E. sinica was set as a sample (sample 1, A); Two samples (samples 2 and 3) were designed by respectively adding the equal ratio mixture of five flavonoids (F) at 50% into alkaloids from S. nux-vomica or E. sinica; Four samples (samples 4-7) were designed by respectively adding F, equal proportion mixture of four saponins (S), equal ratio mixture of six volatile oils/aqueous (V), and equal proportion mixture of six polysaccharides (P) at 50% into A. Murine macrophage cells and chondrocytes were exposed to the new recipes, and then the half inhibitory concentration (IC₅₀) of recipes for inhibiting (PGE₂) in macrophages and the half effective concentration (EC₅₀) for promotion of the secretion of IL-6, IL-1β, and TNF-α in macrophages were detected and compared. The interactions among the active fractions were evaluated by comparing the experimental EC₅₀/IC₅₀ values to their corresponding additive EC₅₀/IC₅₀ values calculated by the least square optimum method. The in vivo pharmacodynamics of the best combination was evaluated by the ear swelling model and delayed type hypersensitivity (DTH) model in mice. Results Sample 4 has good comprehensive activities of cellular anti-inflammation and immunoregulation. Moreover, strong synergistic effect among the effective fractions in this sample was observed; Cellular anti-inflammatory activities of samples 5, 6, and 7 were equivalent with sample 4; Samples 5 and 7 had good comprehensive cellular immunoregulation activity; But the alkaloids mixture (A) and the combinations of S. nux-vomica or E. sinica alkaloids with F (samples 2 and 3) were significantly weaker than sample 4 in cellular anti-inflammation, immunoregulation, and chondrocyte-proliferation. Sample 4 also exhibits a certain effect on in vivo anti-inflammation and immunity in mice when A was decreased at 25% or 5%. Conclusion It is not suitable to design a combination just by alkaloids from S. nux-vomica and alkaloids of E. sinica; Alkaloids from S. nux-vomica or alkaloids of E. sinica are also not appropriate to solely combine with the mixture of flavonoids (F). When A is combined with F, S, V, and P, respectively, synergistic or additive effects among the active fractions are usually observed. These active fractions help to strengthen comprehensive cellular pharmacological activities of A. Sample 4 not only has good cellular activities of anti-inflammation and immunoregulation but also has better in vivo effect on anti-inflammation and immunity, suggesting that it is feasible to screen the optimized Chinese medicine formula based on cellular pharmacological experiments.

上海市科学技术委员会支撑项目(13401901100)