中药挥发油/水提物的细胞抗炎、免疫及骨细胞修复活性的比较

Comparison of Anti-inflammatory,Immunoregulatory and Chondrocyte-proliferative Activities of Chinese Medicine Volatile Oil/Aqueous Extracts

目的以腰痛宁衍生方为基础比较桂枝、土鳖虫等6类中药挥发油/水提物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。方法在马钱子、麻黄生物碱+甘草等5种药材和黄酮+甘草等4种药材皂苷混合物的基础上,分别加入50%的苍术、乳香+没药、独活、桂枝挥发油、50%的土鳖虫、全蝎+僵蚕水提液组成新的中药有效部位组方。测定各样品抑制小鼠巨噬细胞(Ana-1)中前列腺素E2(PGE2)增殖的半数抑制浓度(IC50)及促进白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子(TNF-α)因子增殖和促进IL-1β诱导的软骨细胞增殖的半数有效浓度(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。结果在6类中药挥发油/水提物有效部位中,桂枝挥发油抑制Ana-1分泌PGE2及促进Ana-1分泌IL-1β、IL-6的活性最强;土鳖虫、僵蚕+全蝎水提液促进Ana-1分泌TNF-α的作用最佳,并在细胞抗炎、促进软骨细胞增殖活性和其他细胞免疫模型中表现出较好的活性。各挥发油/水提物有效部位促进IL-1β诱导的软骨细胞增殖的活性相当。结论桂枝挥发油具有良好的抗炎、免疫及促软骨细胞增殖等综合药理活性,可作为风湿骨病处方的优选有效部位并用其替代腰痛宁处方中的乳香+没药挥发油以降低乳香+没药引起的不良反应。腰痛宁组方药材中虫类药材的水提物也可作为风湿骨病处方的候选有效部位。

Objective To compare the effects of six Chinese medicine effective fractions of volatile oils and aqueous extracts on cell anti-inflammation, immunoregulatory and proliferative activities based on a Yaotongning（YTN） derivative formula, thus to supply evidence for screening candidate medicines of rheumatoid bone diseases. Methods Volatile oils（extracted from Rhizoma Atractylodis,Olibanum,Myrrha,Radix Angelicae Pubescentis,or Guizhi）or aqueous extracts（extracted from Tubiechong,Scorpio and Bombyx Batryticatus）at 50 % were added into the mixture of alkaloids（extracted from Semen Strychni andHerba Ephedrae） and flavonoids（extracted from Radix Glycyrrhizae and other four herbs）to compose six effective fractionrecipes. Murine macrophage cells and chondrocytes were exposed to the six recipes, and then their half effective inhibitory concentration（IC50） for PGE2 and the half effective concentrations （EC50）to IL-6, IL-1β, TNF-α in macrophages as well as EC50 to IL-1β-induced proliferation of chondrocytes were detected. 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 optimization method. Results Among the six investigated volatile oil/aqueous extract effective fractions, the volatile oils of Guizhi had the best effect on inhibiting the production of PGE2 and promoting the secretion of IL-1β and IL-6 in macrophages. The aqueous extracts of Tubiechong, Scorpio and Bombyx Batryticatushad the best effect on stimulating TNF-α factor in macrophages and had good activity in cell inflammation and other cell immunoregulatory models. The volatile oils and aqueous extracts active fractions had similar activities on promoting the secretion of chondrocytes induced by IL-1β. Conclusion The volatile oils of Guizhi possesses good cell anti-inflammation, immunoregulatory and proliferative activities and can be the preferred candidate effective fractions of rheumatism prescription. It can replace the volatile oils of Olibanum and Myrrhain YTN prescription to reduce adverse reaction.The aqueous extracts of insect medicinal materials in YTN prescription can also be candidate effective fractions in rheumatism prescription.