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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 investigate the effect of saponin (SAAB) on the activation of mast cells. Methods The peritoneal mast cells (RPMC) were obtained in vitro and the toxic effects of SAAB on RPMC were observed by MTT. Anti-DNP Ig E was used to induce RPMC degranulation. The effect of SAAB on degranulation rate was observed. The histamine release The release of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) from mast cells was detected by enzyme-linked immunosorbent assay. Serotonin and threonine protein kinase K ) Phosphorylation, TNF-alpha and IL-6 expression. Results SAAB had no toxic effect on RPMC at 25 ~ 100μg / L. In vitro anti-DNP Ig E significantly increased the release of histamine, TNF-α and IL-6, and induced the phosphorylation of Akt and TNF-α and IL-6 (200μg / L) and medium (100μg / L) could significantly inhibit the release of histamine, TNF-α and IL-6, inhibit the phosphorylation of Akt and decrease the synthesis of TNF-α and IL-6 (P <0.05). Conclusion TBI regulates allergic inflammation by reducing the synthesis and release of inflammatory cytokines.