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目的:以腰痛宁中生物碱加黄酮有效部位拆方为基础比较不同中药黄酮有效部位对细胞抗炎、免疫及骨细胞修复的影响,为风湿骨病处方候选药物筛选有效部位提供依据。方法:在腰痛宁马钱子、麻黄生物碱基础上,分别加入50%的甘草黄酮、淫羊藿黄酮、红花黄酮、骨碎补黄酮、桑寄生黄酮以及5种黄酮的等比例混合物,得到6个有效部位组方。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Ana-1细胞分泌白细胞介素1β(IL-1β),白细胞介素-6(IL-6),肿瘤坏死因子-α(TNF-α)炎症因子的促进作用;对脂多糖(LPS)诱导的Ana-1细胞释放前列腺素E2(PGE2)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:骨碎补黄酮为主的组方促进IL-1β分泌的活性最强;红花黄酮为主的组方促进IL-6分泌的活性最强且其他活性仅次于最佳的组方;由马钱子、麻黄生物碱+5种黄酮混合物构成的组方抑制PGE2分泌及促进TNF-α分泌、促进软骨细胞增殖的活性最强,且各模型下该组方中有效部位间有极强的协同作用。促软骨细胞增殖模型下,单一黄酮有效部位与生物碱构成的组方中的有效部位之间均表现出较强的拮抗作用。结论:不同药材黄酮之间有出色的协同增效作用,红花黄酮、骨碎补黄酮及各药材黄酮混合物具有良好的抗炎、免疫及骨细胞修复等综合药理活性,提示红花黄酮、骨碎补黄酮及不同药材黄酮混合物可作为风湿骨病药物的优选有效部位。
OBJECTIVE: To compare the effect of different active fractions of flavonoids from different Chinese traditional medicines on cellular anti-inflammatory, immunological and repair of bone cells based on the prescription of alkaloids and flavones in Yaotongning Decoction, and to provide the basis for screening valid candidate sites for rheumatoid bone diseases. Methods: Fifty percent of licorice flavonoids, Epimedium flavanoids, safflower flavonoids, flavonoids of Radix isatidis, flavonoids of Radix Isatidis and five kinds of flavonoids were added into the mixture of 50% 6 effective parts of the group side. Each sample was evaluated for the secretion of interleukin 1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and IL-12 by macrophage Ana-1 cells by using EC50 or IC50 values α (TNF-α); Inhibitory effect of Lipopolysaccharide (LPS) -induced prostaglandin E2 (PGE2) -induced Ana-1 cells; and IL-1β-induced chondrocyte proliferation. At the same time, the least square method was used to calculate the EC50 or IC50 superposition value of each sample. The interaction between the effective parts was analyzed according to the difference between the EC50 or IC50 superposition value and the experimental value. Results: The preparation of rhubarb-flavonoids-based group was the most active in promoting the secretion of IL-1β; the safflower-flavonoid-based group had the strongest activity of promoting the secretion of IL-6 and the other activity was the second best. The combination of Strychnos nux-vomica, ephedrine alkaloids and flavonoids of five flavones inhibited the secretion of PGE2, promoted the secretion of TNF-α, promoted the proliferation of chondrocytes, and had the highest level of active sites in each group Synergy. Under the model of chondrocyte proliferation, all the active sites in the active site of single flavonoids and alkaloids showed strong antagonism. Conclusion: The flavonoids of different herbs have excellent synergistic effect. The flavonoids of safflower flavonoids, flavonoids and various medicinal materials have good pharmacological activities such as anti-inflammation, immunology and repair of bone cells, suggesting that safflower flavonoids and bone Broken-up flavonoids and flavonoids mixture of different herbs can be used as rheumatoid osteoporosis drugs effective sites.