芝麻素通过PKCα/ NF-κB 信号途径抑制肥大细胞活化

目的:观察芝麻素对肥大细胞活化和炎症介质释放的影响并探讨其可能的作用机制。方法:体外培养HMC-1细胞,用10 μg/ ml 的anti-DNP IgE 处理细胞6 h 后,再用100 ng/ ml 的DNP-HAS 孵育10 min 诱导HMC-1 细胞活化,在DNP-HAS 处理前给予最终浓度25、50 和100 μg/ L 芝麻素预处理30 min 后光镜下观察芝麻素对肥大细胞脱颗粒的影响,ELISA 法检测芝麻素对肥大细胞组胺、TNF-α、IL-6、IL-1β、IL-8 释放的影响,Western blot 方法观察芝麻素对Fc RI 受体下游信号Lyn 和 Syk,PKC琢激活,IκB琢磷酸化和NF-κB 活化的影响。结果:DNP-HAS 能明显诱导HMC-1 细胞脱颗粒,促进组胺和TNF-α、IL-6、IL-1β和IL-8 等细胞因子的释放,激活Fc着RI 受体下游信号分子Lyn、Syk 和PKCα,磷酸化IκBα,促进NF-κB 活化(P<0.05)。芝麻素高(100 μg/ L)、中(50 μg/ L)浓度能明显抑制HMC-1 细胞脱颗粒和炎症介质的释放,阻断Fc着RI 受体下游信号激活,抑制NF-κB 活化(P<0.05)。结论:芝麻素通过PKCα/ NF-κB 途径抑制肥大细胞活化和炎症介质释放。

Sesamin suppresses mast cell activation through inhibition of PKCα/ NF-κB signaling pathway

Objective:To investigate the effect of sesamin on mast cell activation and its inflammatory mediator release,as well as its possible mechanisms of action.Methods: HCM-1 cells were activation by stimulation with 10 μg/ ml anti-DNP IgE for 6 h and challenge with 100 ng/ ml DNP-HAS for 10 min.Sesamin was administration at the concentration of 25,50 and 100 μg/ L prior to DNP-HAS challenge,subsequently the effect of sesamin on mast cell degranulation was investigated by light microscope,and histamine release and expression of cytokines such as TNF-αIL-6,IL-1β,IL-8 of mast cells after sesamin treatment were investigated by ELISA.Western blot was used to determine the effect of sesamin on Fc RI downstream signaling including Lyn,Syk and PKCα activation,and IκBαphosphorylation and NF-κB activation.Results: DNP-HAS significantly increased mast cell degranulation,histamine release and those cytokines expression,enhanced Lyn,Syk,PKCα, IκBαphosphorylation and NF-κB activation(P <0.05). Sesamin(50,100 μg/ L) significantly decreased mast cell degranulation,histamine release and cytokines expression (TNF-α,IL-4,IL-1β,and IL-8),reduced activity of Lyn,Syk,kinases and PKCαand IκBαphosphorylation,and inhibited NF-κB activation(P<0.05). Conclusion: Sesamin suppresses mast cell activation and inflammatory mediators release through inhibition of PKCα/ NF-κB signaling pathway.