热毒宁注射液速发型过敏反应机制研究

Mechanism of rapid allergic reaction induced by Reduning injection

目的探讨热毒宁注射液诱导的速发型过敏反应的特点,并初步探究其致敏机制。方法以大鼠嗜碱性细胞白血病细胞(RBL-2H3)作为热毒宁注射液诱发速发型过敏反应的评价模型。采用中性红染色法观察致敏时细胞脱颗粒的形态变化,测定β-氨基己糖苷酶(β-hex)的释放率来反映细胞的脱颗粒程度,MTT法评价致敏时的细胞活力;ELISA法测定白介素-4(IL-4)含量的变化,Fluo-3 AM Ca^(2+)荧光探针探究Ca^(2+)荧光强度的变化。结果热毒宁注射液可以引起RBL-2H3脱颗粒,剂量依赖性地增加细胞内β-hex以及IL-4的释放率,但是对细胞活力并没有影响;同时,热毒宁注射液作用于RBL-2H3后,Fluo-3 AM Ca^(2+)荧光探针的荧光强度先增加后减弱,细胞内Ca^(2+)浓度呈现先增加后减少的趋势。结论热毒宁注射液通过调节细胞内Ca^(2+)的浓度引起RBL-2H3脱颗粒,诱导β-hex以及IL-4等相关炎性介质的产生,从而引发速发型过敏反应。

Objective To determine the characteristics and allergy mechanism of rapid allergic reactions induced by Reduning injection.Methods Rat basophil leukemia cells(RBL-2H3)was used as the evaluation rat model of anaphylaxis induced by Reduning injection.Neutral red staining was used to observe the morphological changes in cell degranulation.The release rate ofβ-hexosaminidase(β-hex)was used to reflect the degranulation degree of the cells.The cell viability was evaluated by MTT assay.The content of IL-4 was measured by ELISA,and the fluorescence intensity of Ca^(2+) was detected by Fluo-3 Am Ca^(2+) fluorescent probe.Results Reduning injection induced the degranulation of RBL-2H3,increased the release rate ofβ-hex and IL-4,but had no effect on the cell viability.When Reduning injection acted on RBL-2H3,the fluorescence intensity of Ca^(2+) increased first and then decreasd,indicating that the intracellular Ca^(2+) concentration increased first and then decreased.Conclusion Reduning injection can induce the degranulation of RBL-2H3 by regulating the concentration of intracellular Ca^(2+),and induce the production of inflammatory mediators such asβ-hex and IL-4,leading to rapid allergic reactions.