补肾活血方有效组分及部分化合物的体外抗炎活性研究

Study on the anti-inflammatory activity of effective components and some compounds in Bushen Huoxue Recipe

目的:观察补肾活血方水提取部位、醇提取部位、总多糖部位以及大黄酸、大黄酚等有效组分与化合物对肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)及一氧化氮(NO)等炎症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:选取补肾活血方不同提取分离部位,以及复方水提液中存在的具有潜在治疗骨关节炎活性的大黄酸、大黄酚化合物,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:复方水提取部位与醇提取部位在100μg/ml,以及大黄酚在20μg/ml和100μg/ml对TNF-α的释放具有显著的抑制;复方醇提取部位在100μg/ml,大黄酸在20μg/ml,以及大黄酚在20μg/ml和100μg/ml对IL-6水平具有显著的抑制;复方水提取部位在100μg/ml、醇提取部位在20μg/ml和100μg/ml、大黄酸在20μg/ml,以及大黄酚在20μg/ml和100μg/ml对NO的释放具有显著抑制。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 water extraction, alcohol extraction, total polysaccharides, rhein and chrysophanol, five effective components and compounds in Bushen Huoxue Recipe on the release concentration of inflammatory mediators tumor necrosis factor alpha （TNF-a）, interleukin-6 （IL-6） and nitric oxide （NO）, and to discuss the preliminary mechanism of compound prescription in treatment of bone arthritis. Methods： Different components of the recipe and compounds, which may have potential effect on osteoarthritis activity were separated from compound recipe. Model of RAW264.74 rat macrophages stimulated in vitro by lipopolysaccharide （LPS） was set up and en- zyme-linked immunosorbent assay （ELISA） was applied to detect the effect of reagents on IL-6, TNF-a and NO released by model cells. Re- suits： water extraction（ 100μg/ml） , alcohol extraction（ 100μg/ml）, and chrysophanol（20μg/ml, 100μg/ml） had a significant inhibition on the release of TNF-a; alcohol extraction（100μg/ml）, rhein（20μg/ml） and chrysophanol（20μg/ml, 100μg/ml） had significant inhibition on IL-6 ; water extraction（ 100μml）, alcohol extraction （20μg/ml, 100μg/ml）, rhein （20μg/ml） and chrysophanol （20μg/ml, 100μg/ ml） had a remarkable inhibition on the release of NO. Conclusions： Bushen Huoxue recipe in treatment of osteoarthritis activity may be probably proformed by inhibiting the release of inflammatory mediators and the effect of TNF-a, IL-6 and NO, which have a great significance in verifying pharmacodynamic activity and exploring material basis of effective components.