丹皮酚对RBL-2H3肥大细胞活化脱颗粒的影响

Inhibition of Paeonol on Activating and Degranulating of RBL-2H3 Mast Cells

目的通过建立体外抑制肥大细胞脱颗粒的药效学模型,探讨丹皮酚抗Ⅰ型变态反应作用机制。方法用MTT比色法检测不同浓度的丹皮酚对RBL-2H3肥大细胞增殖的影响。通过ELISA法,明确体外RBL-2H3细胞活化分泌组胺及TNF-α动力学特征。检测不同浓度药物对RBL-2H3肥大细胞分泌组胺及TNF-α的影响。结果丹皮酚对RBL-2H3细胞增殖有抑制作用,IC50值为0.22 mg/mL。RBL-2H3细胞活化脱颗粒,分泌组胺的释放率在前30 m in上升较快,30 m in后上升进入平台期;TNF-α分泌的量在1h达高峰。丹皮酚抑制RBL-2H3细胞释放组胺和TNF-α作用不弱于色甘酸钠0.1,0.5 mg/mL。丹皮酚浓度的对数与其对RBL-2H3细胞释放组胺和TNF-α的抑制率呈线性相关(P=0.000<0.01)。结论丹皮酚呈剂量依赖性抑制RBL-2H3细胞分泌组胺和TNF-α。

Objective To study the paeonol direct effect on the degranulation function of RBL -2H3 mast cells. Methods Paeonol of different concentration influenced the proliferation of RBL-2H3 mast cells determined by MTT colorimetric assy. Established a pharmic model on the degranulation of RBL-2H3 mast cells, activated by compound 48/80 （ 10 μg/mL）. Pretreated RBL-2H3 cells with different dilutions of paeonol,The histamine and tumor necrosis factor-alpha（TNF-α） from the supernatants, the spontaneously released histamine and the total histamine were all determined by ELISA. Results Paeonol of different concentration suppressed the proliferation of RBL-2H3 mast cells. The pinnacle of the histamine released from RBL-2H3 mast cells was （ 23.14 ± 1.01 ） % at 30min. The pinnacle of the TNF-α released from RBL-2H3 mast cells was （67.65 ±5.33 ） pg/mL at 60min. The suppressed percentage of the histamine and TNF-α released from RBL-2H3 mast cells pretreated by paeonol was no less than that of sodium cromoglicate control group,and in a concentration dependent manner upon pretreating with paeonol. Conclusion Paeonol may suppress degranulation of RBL-2H3 cell in a direct way,which could be one of the mechanism of anti-anaphylactic reaction