目的 评价腰痛宁胶囊中不同活性部位组合的抗炎、免疫及活血活性,以及活性部位间的相互作用.方法 按照腰痛宁组方原则、组合化学思想及各活性部位在腰痛宁中的重要性,在马钱子与麻黄总生物碱基础上依次加入腰痛宁组方药材的总黄酮、总皂苷、挥发油和多糖等活性部位,得到由不同活性部位混合组成腰痛宁拆方、组方及全方共计6个样品.运用脂多糖(LPS)诱导Ana-1细胞释放前列腺素E₂(PGE₂)模型,有丝分裂原(ConA)诱导脾淋巴细胞产生白细胞介素-2(IL-2)模型及刺激人脐静脉内皮细胞释放NO模型,测定各样品抑制PGE₂产生的半数抑制浓度(IC₅₀),促进IL-2和NO分泌的半数有效浓度(EC₅₀);同时采用最小二乘优化方法,计算各样品的IC₅₀/EC₅₀叠加值,将其与IC₅₀/EC₅₀实验值进行比较,分析不同模型下各活性部位间的相互作用类型.结果 6个样品均有抑制PGE₂产生及促进IL-2分泌的活性,但只有由总生物碱、总黄酮和总皂苷组成的样品III有刺激人脐静脉内皮细胞释放NO的EC₅₀,且样品III在以PGE₂、IL-2及NO为指标的抗炎、免疫及活血模型中均表现出优于其他样品的活性,不同的活性部位组合在这3个模型下产生的相互作用类型不同.只有样品III中的总生物碱、总黄酮及总皂苷3个活性部位在3个模型下均存在协同增效作用.结论 腰痛宁拆方、组方及全方样品对Ana-1细胞中PGE₂的产生均表现出良好的抑制作用、对小鼠脾淋巴细胞分泌IL-2也表现出良好的促进作用,显示出腰痛宁处方的合理性、科学性及优良的抗炎和免疫活性.腰痛宁中总生物碱、总黄酮与总皂苷间的协同增效作用使腰痛宁拆方样品III在抗炎、免疫与活血方面的活性均优于其他样品,表明腰痛宁组方具有优化空间.可望以样品III为基础精简腰痛宁组方,开发高效、质量易控的治疗风湿痹病新药.

Objective To evaluate the in vitro effects of different combinations of the active fractions in Yaotongning Capsule (YTNC) on anti-inflammation, immunity, and activating blood circulation, and the interactions among these active fractions. Methods Six samples were prepared by adding flavonoids, saponins, volatile oils/aqueous extracts, and polysaccharides, which were extracted from the material medicines in YTNC, to the alkaloids of Strychnos nux-vomica (Ma Qian Zi) and Ephedra sinica (Ma Huang) in turn according to the formulating principle of YTNC, combination chemistry concept, and their importance in the formula. The effects of the six samples on the production of lipopolysaccharide (LPS)-induced prostaglandin E₂ (PGE₂) in Ana-1 cells, concanavalin (ConA)-induced interleukin-2 (IL-2) in splenocyte, and nitrogen monoxide (NO) in human umbilical vein endothelial cells (HUVEC) were explored by detecting their half-maximal inhibitory concentration (IC₅₀) or half-maximal effective concentration (EC₅₀). An innovative method based on least square optimization was applied to calculate the additive IC₅₀/EC₅₀ value of each sample. The interactions of the active fractions in these samples were investigated by comparing the additive IC₅₀/EC₅₀ values with the corresponding experimental IC₅₀/EC₅₀ values. Results All the six samples could inhibit the production of PGE₂ and promote the secretion of IL-2 in the investigated concentration range. Only sample III consisting of the alkaloids, flavonoids, and saponins from YTNC had the EC₅₀ value of promoting NO production. Moreover, the sample also displayed excellent activities in the three models of LPS-induced PGE₂ production, ConA-induced IL-2 production, and NO production, which could reflect the capability of anti-inflammation, enhancing the body's immunity, and activating blood circulation, respectively. Different combination of the active fractions from YTNC produced the different interaction types in the three models. And the active fractions in sample III, i.e., the alkaloids, flavonoids, and saponins from YTNC, generated high synergistic effects in all the three models. Conclusion All the six samples derived from the different combination of the active fractions from YTNC show the good activity of inhibiting PGE₂ production and promoting IL-2 secretion. Therefore, the YTNC formula is reasonable and has good activity of anti-inflammation and immunity. However, the YTNC formula still could be improved by reducing the categories of its active fractions because the combination of the alkaloids, flavonoids, and saponins exerted better activities than the other samples and the three kinds of active fractions produced an excellent synergistic effect in all the three models. Synthetically evaluating various activities of YTNC in the present work may lead to develop new Chinese materia medica for rheumatic arthralgia treatment, which is more effective and easily controllable in quality.

上海市科技支撑项目(13401901100)