痹祺胶囊提取物对RAW264.7细胞模型的抗炎作用

Anti-inflammatory effect of three extractions from Biqi Capsules on LPS-stimulated RAW264.7 macrophages

目的通过比较研究痹祺胶囊(马钱子、党参、丹参、白术、茯苓、川芎、三七、地龙、甘草、牛膝)不同溶剂提取物的抗炎作用,从而确定最佳提取溶剂。方法 3种溶剂提取痹祺胶囊,水、石油醚、正丁醇的提取物与脂多糖(LPS)刺激小鼠单核/巨噬细胞RAW264.7共孵,用酶联免疫吸附法(ELISA)、Griess Reagent法检测细胞分泌的肿瘤坏死因子(TNF-α)和一氧化氮(NO)的水平;以实时定量逆转录-聚合酶链式反应(Real-time RT-PCR)检测细胞炎症相关指标白介素-6(IL-6)、环氧合酶-2(COX-2)、诱导型一氧化氮合酶(iNOS)、TNF-α的mRNA表达水平,比较3种提取物的抗炎作用。结果 3种提取物均能显著抑制LPS刺激的RAW264.7细胞分泌NO和TNF-α,其中水提取物的抑制作用最强,且剂量依赖性最显著。在基因表达水平,只有水提取物对LPS刺激的RAW264.7细胞的IL-6、COX-2、iNOS的mRNA表达有显著抑制作用。结论体外细胞药理学实验结果表明,痹祺胶囊内容物以水、石油醚、正丁醇3种溶剂体系提取得到提取物均具有抗炎作用,但水提取物抗炎作用更显著。

AIM To determine the best extraction of Biqi Capsules （Strychni Semen, Codonopsis Radix, Salviae miltiorrhizae Radix et Rhizome, Atractylodis macrocephalae Rhizoma, Poria, Chuanxiong Rhizoma, Notoginseng Radix et Rrhizoma, Pheretima, Glycyrrhizae Radix et Rhizoma, Achyranthis bidentatae Radix） by different solvents, and to compare their anti-inflammatory effects. METHOD The anti-inflammatory action of aqueous, petroleum ether and n-butanol extracts to lipopolysaccharide （LPS） -stimulated mouse mononuclear macrophage cells line RAW264.7 were compared in vitro. Production of nitric oxide （NO） and tumor necrosis factor-alpha （TNF-α） were measured by the Griess colorimetric method and Enzyme-linked Immuno Sorbent Assay （ELISA）, respectively. The mRNA expression levels of inducible nitric oxide synthase （ iNOS）, cyclooxygenase （ COX-2）, TNF-α, and interleukin-6 （IL-6） were detected by quantitative real-time RT-PCR. RESULTS Three extracts of Biqi Capsules significantly reduced the levels of NO and TNF-α production in LPS-stimulated RAW264.7 （ P 〈 0. 01 ）, and the effect of aqueous extract of Biqi Capsules had the most effective anti-inflammatory action than that of petroleum ether extract and n-butanol extract in the dose-dependent manner. The results of real-time RT-PCR showed that iNOS, COX-2 and IL-6 mRNA expression decreased. CONCLUSION These results demonstrate that all three Biqi Capsule extracts have anti-inflammatory effects, with aqueous extract being the most effective.