知母皂苷对肥大细胞活化的影响

Effects of saponins from Anemarrhena asphodeloides Bunge on mast cells activation

目的探讨知母皂苷（SAAB）对肥大细胞活化的影响。方法体外获取大鼠腹腔肥大细胞（RPMC）,MTT观察SAAB对RPMC毒性作用;利用Anti-DNP Ig E诱导RPMC脱颗粒,观察SAAB对RPMC脱颗粒率的影响;酶联法检测肥大细胞组胺释放;酶联免疫吸附测定法检测肥大细胞肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）释放;Western blot免疫印迹检测丝氨酸/苏氨酸蛋白激酶蛋白激酶B（Akt）磷酸化、TNF-α和IL-6的表达。结果 SAAB在25~100μg/L对RPMC没有毒性作用,体外anti-DNP Ig E明显促进RPMC组胺,TNF-α和IL-6的释放,诱导Akt磷酸化和提高TNF-α和IL-6的合成,SAAB高（200μg/L）、中（100μg/L）浓度明显抑制肥大细胞组胺,TNF-α和IL-6的释放,并抑制Akt磷酸化和降低TNF-α和IL-6的合成（P〈0.05）。结论知母皂苷通过降低肥大细胞炎症相关因子合成和释放调控过敏性炎症。

Objective To explore the effect of saponins from Anemarrhena asphodeloides Bunge（ SAAB） on mast cell activation.Methods Rat peritoneal mast cells（ RPMC） were isolated from SD rats. The cytotoxicity of SAAB was determined by MTT. The effect of SAAB on mast cell activation and degranulation was investigated by anti- DNP Ig E. Histamine release from RPMC was measured by EIA. TNF- α and IL- 6 release from RPMC was measured by ELISA. The proteins expression of Akt and TNF- α,IL- 6 were measured by Western blotting. Results SAAB had no cytotoxicity on RPMC between 20 μg / L and 100 μg / L,Ig E-mediated RPMC histamine and TNF- α,IL- 6 release were significantly increased. Furthermore,Akt phosphorylation and TNF- α,IL- 6 expressions were significantly increased. SAAB（ 100 and 50 μg / L） inhibited anti- DNP Ig E- mediated histamine and TNF- α,IL- 6 release from RPMC. Akt phosphorylation and TNF- α,IL- 6 expressions were significantly decreased by SAAB（ 100 and 50 μg / L） in RPMC（ P 〈 0. 05）. Conclusion All these results indicated that SAAB regulated allergic inflammation through decrease mast cells- mediated inflammation factors production and release.