补肾活血方中4种有效成分对炎性因子释放抑制活性研究

Research on activity inhibition of 4 effective components in Bushen Huoxue Formula on release of inflammatory cytokines

目的:观察补肾活血方中异欧前胡素、齐墩果酸、蜕皮甾酮、虎杖苷等4种有效成分对肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)及一氧化氮(NO)等炎性介质释放浓度的影响作用,探讨复方治疗骨关节炎的初步机制。方法:选取补肾活血方水提液中存在的具有潜在治疗骨关节炎活性的异欧前胡素等4种化合物,采用体外脂多糖(LPS)刺激的RAW264.74小鼠巨噬细胞为模型,利用酶联免疫法(ELISA)测定试药对模型细胞释放TNF-α、IL-6及NO的抑制作用。结果:异欧前胡素在100μg/m L,对TNF-α的释放有显著抑制作用(P<0.01);异欧前胡素在20、100μg/m L,虎杖苷在100μg/m L对IL-6水平有显著抑制(P<0.01);异欧前胡素与齐墩果酸在20、100μg/m L,蜕皮甾酮与虎杖苷在100μg/m L,对NO的释放具有显著抑制(P<0.01)。结论:补肾活血方的治疗骨关节炎活性可能是通过抑制炎症介质释放,降低TNF-α、IL-6、NO等含量发挥作用,这对验证复方药效活性,探究效应物质基础具有重要意义。

Objective： To observe the influence of 4 effective components in Bushen Huoxue Formula such as isoimperatorin, oleanolic acid, ecdysterone, and polydatin on the concentration of inflammatory mediators as tumour necrosis factor-alpha （TNF- α ）, interleukin-6 （IL-6） and nitric oxide （NO）, in order to discuss the accumulative mechanism of treating osteoarthritis with compound prescriptions. Methods： 4 compounds existed in water-extraction of Bushen Huoxue Formula, which might have the potential effect on osteoarthritis activity were separated from compound formula. Model of RAW264.74 mouse macrophages stimulated in vitro by lipopolysaccharide （LPS） was established and enzyme-linked immunosorbent assay （ELISA） was applied to detect the inhibition effect of test medicines on IL-6, TNF- α and NO released by model cells. Results： Isoimperatorin （100μg/mL） had a significant inhibition on the release of TNF- α ; isoimperatorin （20, 100μg/mL） and polydatin （100μg/mL） had significant inhibition on the release of IL-6; isoimperatorin （20, 100μg/mL）, oleanolic acid （20, 100μg/mL）, ecdysterone （100μg/mL）, and polydatin （100μg/mL） had a remarkable inhibition on the release of NO. Conclusion： Bushen Huoxue Formula could be used to treat osteoarthritis activity, and the mechanism might be related to the decreasing on the contents of TNF- α, IL-6 and NO through inhibiting the release of inflammatory mediators, which had a great significance in verifying pharmacodynamic activity and exploring substantial basis of compound prescriptions.