聚山梨酯80诱发类过敏反应机制研究  

Study on mechanisms of anaphylactoid reaction induced by Tween 80

作　　者：李振虎[1,2] 王化龙[1,2] 刘艳庭[1,2] 高秀梅[1] 徐砚通[1,2] 

机构地区：[1]天津中医药大学天津市现代中药重点实验室,天津300193 [2]天津国际生物医药联合研究院中药新药研发中心,天津300457

出　　处：《中国新药杂志》2016年第23期2664-2669,共6页Chinese Journal of New Drugs

基　　金：国家"重大新药创制"科技重大专项资助项目(2013ZX09201020;2012ZX09101212);天津市高等学校"创新团队培养计划"资助项目(TD2-5031)

摘　　要：目的:研究聚山梨酯80诱发类过敏反应的可能作用机制。方法:采用小鼠耳廓蓝染模型考察聚山梨酯80对血管通透性的影响,验证聚山梨酯80诱发类过敏反应效应;采用RBL-2H3细胞脱颗粒模型探索聚山梨酯80诱发类过敏反应的可能作用机制。首先检测β-氨基己糖苷酶释放,考察聚山梨酯80对过敏介质释放的影响;进而采用细胞膜特异荧光染料FM4-64在高内涵系统考察聚山梨酯80对细胞脱颗粒囊泡回收的影响;最后采用荧光偏振法考察聚山梨酯80对细胞膜流动性影响。结果:小鼠耳廓蓝染实验证明聚山梨酯80能够显著提高血管通透性,证实了聚山梨酯80诱发类过敏反应效应;在RBL-2H3细胞模型上,β-氨基己糖苷酶释放实验证实聚山梨酯80能够显著增加β-氨基己糖苷酶释放,表明聚山梨酯80能够直接诱发过敏介质释放;高内涵实验显示聚山梨酯80能够显著促进囊泡回收,提示聚山梨酯80能够促进细胞脱颗粒过程中的囊泡循环;荧光偏振法检测实验证实聚山梨酯80显著降低相对荧光偏振度,提示聚山梨酯80能够提高细胞膜流动性。结论:研究结果提示聚山梨酯80诱导类过敏反应发生机制可能通过提高细胞膜流动性加速了囊泡循环,促进了细胞脱颗粒,增加了过敏介质释放引起类过敏反应症状。Objective： To investigate the possible mechanisms of anaphylactoid reaction（ AR） induced by Tween 80. Methods： To confirm the AR effects of Tween 80 in vivo,the model of mouse ear staining by Evans blue was adopted to evaluate vascular permeability. To investigate the possible action mechanisms of Tween 80 on AR,the model of degranulation of RBL-2H3 cells was used. The effect of Tween 80 on the release of allergic mediators was tested by β-hexosaminidase assay. The effects of Tween 80 on the recycling of vesicle membrane in the cellular degranulation was evaluated by a high content assay（ HCA） using specific membrane labeling fluorescent dye FM4-64. The effect of Tween 80 on membrane fluidity was tested by the membrane fluorescence polarization test（ MFPT）. Results： Mouse ear staining by Evans blue test showed that Tween 80 significantly increased vascular permeability,and it demonstrated the effects of Tween 80 on AR in vivo. On the model of degranulation of RBL-2H3 cells, β-hexosaminidase assay indicated that Tween 80 significantly increased the release of β-hexosaminidase,and it suggested that Tween 80 could induce the release of allergic mediators. The HCA showed that Tween 80 significantly increased the recycling of vesicle membrane,and it suggested that Tween 80 could enhance the recycling of vesicle membrane during cellular degranulation. The MFPT displayed that Tween 80 significantly decreased the relative membrane fluorescence polarization,and it suggested that Tween 80 could increase the fluidity of cell membrane. Conclusion： Tween 80 may induce AR by increasing membrane fluidity,enhancing vesicle recycle of cellular degranulation,increasing the release of allergic mediators,and finally resulting in allergic symptoms.

关 键 词：聚山梨酯80 类过敏反应 细胞脱颗粒 细胞膜流动性 

分 类 号：R285[医药卫生—中药学]