盐酸青藤碱引发类变态反应的机制研究

Study on the mechanism of sinomenine hydrochloride induced pseudo-allergic reaction

目的研究盐酸青藤碱致类变态反应的作用机制,为减轻盐酸青藤碱类药物制剂在临床应用中的不良反应提供依据。方法给小鼠脚掌皮下注射不同剂量的盐酸青藤碱,观察其类变态反应情况,并采用苏木精-伊红(HE)染色分析产生类变态反应时的皮肤组织形态变化,利用肥大细胞Avidin染色考察盐酸青藤碱对小鼠脚掌皮肤肥大细胞的激活作用。小鼠尾静脉注射不同剂量的盐酸青藤碱后,用质谱法检测血清中组胺的含量,用Elisa法检测血清中前列腺素D2(PGD2)、5-羟色胺(5-HT)、单核细胞趋化蛋白-1(MCP-1)、肿瘤坏死因子-α(TNF-α)和白细胞介素-8(IL-8)的含量。结果皮下注射盐酸青藤碱可使小鼠脚掌肿胀及组织液渗出(P<0.05),HE染色可见明显的血管扩张现象,Avidin染色显示盐酸青藤碱可刺激皮肤组织肥大细胞脱颗粒。盐酸青藤碱高剂量组可使小鼠血清中组胺质量浓度达到173.83±26.25 ng·mL^(-1),同时PGD2、5-HT、MCP-1、TNF-α和IL-8的含量也显著升高(P<0.05)。结论盐酸青藤碱可剂量依赖性激活肥大细胞脱颗粒,使小鼠血清中致敏介质水平升高,从而引发类变态反应。

Objective To clarify the mechanism of sinomenine hydrochloride induced pseudo-allergic reaction,and to provide experimental basis for the safe use of sinomenine hydrochloride in clinical application.Methods The pseudo-allergic reaction of sinomenine hydrochloride was evaluated by a mouse model of hindpaw extravasation in vivo.Hematoxylin-eosin(HE)staining was applied for studying the histological changes by sinomenine hydrochloride on mouse hindpaw skin tissue,and mast cell.Avidin staining was carried out to investigate the activation effect of sinomenine hydrochloride on mouse hindpaw skin mast cells.Histamine serum release levels was determined by mass spectrometry method.The release levels of prostaglandin D2(PGD2),5-hydroxytryptamine(5-HT),monocyte chemotactic protein-1(MCP-1),tumor necrosis factor-α(TNF-α)and interleukin-8(IL-8)were detected by enzyme immunoassay(Elisa).Results Subcutaneous injection of sinomenine hydrochloride triggered mice hindpaw thickness increase and tissue fluid exudation dose-dependently(P<0.05).HE staining showed obvious vasodilation,and Avidin staining showed mast cell degranulation by sinomenine hydrochloride.The serum histamine was up to 173.83±26.25 ng·mL-1 with sinomenine hydrochloride high dose group,and the contents of PGD2,5-HT,MCP-1,TNF-αand IL-8 in the mice serum were increased significantly(P<0.05).Conclusion Sinomenine hydrochloride can dose-dependently trigger mast cell degranulation and increase the level of inflammatory substances in mice serum,and then induce pseudo-allergic reaction.