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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 effect of four effective components of isoimperatorin, oleanolic acid, ecdysterone and polydatin on tumor necrosis factor-α (TNF-α), interleukin-6 ) And nitric oxide (NO) and other inflammatory mediators release of the role of concentration, to explore the initial mechanism of compound treatment of osteoarthritis. Methods: Four kinds of compounds, such as isoimperatorin, have potential therapeutic effect on osteoarthritis in the aqueous extract of Bushen Huoxue Recipe. RAW264.74 macrophages stimulated by lipopolysaccharide (LPS) Enzyme-linked immunosorbent assay (ELISA) was used to determine the inhibition of TNF-α, IL-6 and NO production in model cells. RESULTS: Isoimperatorin had a significant inhibitory effect on the release of TNF-α at 100 μg / mL (P <0.01), with isoimperatorin at 20 and 100 μg / mL and at a dose of 100 μg / mL IL-6 was significantly inhibited (P <0.01); isoimperatorin and oleanolic acid at 20,100μg / mL, ecdysterone and polydatin at 100μg / mL significantly inhibited NO release (P <0.01). Conclusion: The treatment of osteoarthritis by Bushen Huoxue Prescription can play a role by inhibiting the release of inflammatory mediators and decreasing the content of TNF-α, IL-6 and NO, which is of great significance for verifying the pharmacological activity of compound and exploring the substance basis of effect.