防己水煎液总生物碱的镇痛抗炎作用及其机制研究

Study on analgesic and anti-inflammatory effects and mechanism of total alkaloids from Stephania tetrandra S. Moore decoction

目的研究防己水煎液总生物碱的镇痛抗炎作用及其作用机制,为进一步明确防己水煎液中水溶性强、极性大的生物碱类成分的药理作用提供科学依据。方法采用732型阳离子树脂富集防己水煎液中总生物碱。通过醋酸致小鼠疼痛扭体法,热板致小鼠疼痛舔足法观察其镇痛作用。通过二甲苯致小鼠耳廓肿胀法,角叉菜胶致大鼠足趾肿胀法观察其抗炎作用。同时结合脂多糖(LPS)诱导的小鼠巨噬细胞RAW264.7细胞模型探讨作用机制,采用噻唑蓝比色法(MTT法)测定细胞毒性,酶联免疫吸附法(ELISA)检测细胞上清液中一氧化氮(NO),肿瘤坏死因子-α(TNF-α),白介素-6(IL-6)的含量,观察释放炎症因子的变化。结果镇痛实验中总生物碱可有效减少小鼠疼痛扭体次数(P<0.01),可提高小鼠热痛舔足痛阈值(P<0.01)。抗炎实验中可减小耳廓肿胀度(P<0.01),减小足趾肿胀度(P<0.01),提高两者抑制率。RAW264.7细胞模型中可降低炎症因子NO、TNF-α、IL-6的释放(P<0.01)。结论防己水煎液总生物碱具有显著的镇痛抗炎作用,其机制可能与减少炎症因子释放有关。

Objective Study on analgesic and anti- inflammatory effects and mechanism of total alkaloids from Stephania tetrandra S. Moore decoction,which provides a scientific basis for further clarify the pharmacological effects of alkaloids with strong soluble and large polar from Stephania tetrandra decoction. Methods Using 732 cation exchange resin enriched total alkaloids from Stephania tetrandra decoction. Analgesic effect was observed through the writhing induced by acetic acid and hot plate in mice.And anti- inflammatory effect was observed by ear edema induced by xylene in mice and rat paw edema induced by carrageenan.Then combined with lipopolysaccharide（ LPS） induced murine macrophage RAW264. 7 cell model investigated the mechanism of anti- inflammatory. Using MTT（ MTT method） tested cytotoxic activity,and enzyme- linked immunosorbent assay（ ELISA） detected content of nitric oxide（ NO）,tumor necrosis factor- α（ TNF- α） and interleukin- 6（ IL- 6） from cell supernatant and observe the effect of the release of inflammatory cytokines. Results In analgesic experiments,total alkaloid could effectively reduce the writhing in mice（ P〈0. 01） and improve pain threshold in mice（ P〈0. 01）. In inflammatory experiments,total alkaloid could reduce ear swelling（ P〈0. 01）,the toe swelling（ P〈0. 01） and develop inhibition rate. RAW264. 7 cell model could reduce the release of the inflammatory cytokines NO,TNF- α,and IL- 6（ P〈0. 01）. Conclusion： Conclusion Total alkaloid from Stephania tetrandra decoction has significant analgesic and anti- inflammatory effects and the mechanism of anti- inflammatory may reduce the release of inflammatory cytokines.