摘要
目的:探讨木犀草素(Luteolin)的抗I型变态反应的作用机制。方法:通过建立DNP-BSA-Ig E激发致敏的大鼠RBL-2H3细胞模型,分别采用MTT法检测不同浓度(5、15、25μmol/L)Luteolin对RBL-2H3肥大细胞活性的影响;ELISA法检测不同浓度Luteolin对RBL-2H3细胞分泌β-hexosa minidase(β-HEX)及细胞因子TNF-α影响;Flou-4AM荧光探针检测细胞内Ca2+浓度变化;Western blot检测AKT,P-AKT的表达。结果:成功建立致敏的细胞模型,低浓度的Luteolin对RBL-2H3细胞活性无明显影响;不同浓度Luteolin刺激RBL-2H3细胞后,对β-HEX和TNF-α的释放抑制作用呈线性相关,且细胞内Ca2+明显减少;Western blot结果显示随着Luteolin浓度的增加AKT磷酸化水平明显下降。结论:Luteolin呈剂量依赖性抑制RBL-2H3肥大细胞脱颗粒,且通过调节细胞内Ca2+浓度与AKT活性参与其中。
Objective： To investigate the effect of Luteolin on the degranulation function of RBL-2H3 mast cells and its mechanism of action. Methods： DNP-BSA-Ig E was used to establish the sensitized cell model in RBL-2H3 cells. Cell viability was exa mined by MTT assay at different concentrations（ 5,15,25 μmol / L） of Luteolin pretreated RBL-2H3 cells. The effect of Luteolin on β-hexosa minidase（ β-hex） and TNF-α was evaluated by ELISA. The change of Ca2 ＋influx was detected by Flou-4AM calcium ion fluorescent probe. The expression of AKT and P-AKT were detected by Western blot. Results： The sensitized cell model was established successfully. Cell viability had not significantly changes stimulated by different concentrations of Luteolin,and a significantly inhibition role showed the release of β-hex and TNF-α from RBL-2H3 cells. The concentration of intracellular Ca2 ＋was significantly decreased. However,the Western blot results showed that the level of Phosphorylation AKT was decreased. Conclusion： Luteolin suppress RBL-2H3 cell degranulation in dose dependence,which may via impacting the Ca2 ＋influx and the activity of AKT.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2016年第8期1153-1156,共4页
Chinese Journal of Immunology
