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目的:通过比较绿原酸和绿原酸完全抗原对未致敏和致敏RBL-2H3细胞脱颗粒的影响,探讨绿原酸的致敏性及其作用机制,为进一步正确评价绿原酸在中药注射剂不良反应中地位和临床合理用药提供一定的参考。方法:采用未致敏RBL-2H3细胞和致敏RBL-2H3细胞模型,观察绿原酸和绿原酸完全抗原对肥大细胞脱颗粒的影响。比色法检测过敏介质组胺和β-氨基己糖苷酶的释放,中性红染色法进行形态学观察,流式细胞法检测Annexin V阳性细胞率。结果:绿原酸和绿原酸完全抗原直接刺激RBL-2H3细胞并未引起肥大细胞发生脱颗粒反应;将RBL-2H3细胞与绿原酸或双黄连致敏血清共孵育致敏后,绿原酸完全抗原再次作用于细胞,可引起细胞组胺释放率、β-氨基己糖苷酶释放率、Annexin V阳性细胞率和中性红染色脱颗粒细胞率均明显增加。结论:绿原酸作为小分子物质,以半抗原形式存在,本身不具有致敏性,但它能与血清蛋白结合形成复合物而产生免疫原性,从而引起过敏反应。绿原酸作为双黄连注射剂的一种有效成分,同时也是双黄连注射剂引起变态反应的一种致敏原。
Objective: To compare the effect of chlorogenic acid and chlorogenic acid complete antigen on degranulation of unsensitized and sensitized RBL-2H3 cells, explore the sensitization of chlorogenic acid and its mechanism of action, in order to further correctly evaluate the effect of chlorogenic acid in Chinese medicine Injections of adverse reactions in the status and clinical rational use of drugs to provide some reference. Methods: The unsensitized RBL-2H3 cells and sensitized RBL-2H3 cell model were used to observe the effect of chlorogenic acid and chlorogenic acid complete antigen on mast cell degranulation. The release of histamine and β-hexosaminidase were detected by colorimetric assay. The morphology was observed by neutral red staining. The rate of Annexin V positive cells was detected by flow cytometry. Results: RBL-2H3 cells directly stimulated by chlorogenic acid and chlorogenic acid did not cause degranulation of mast cells. After incubation of RBL-2H3 cells with sensitized serum of chlorogenic acid or Shuanghuanglian, Acid complete antigen re-act on the cells, can cause cell histamine release rate, β-hexosaminidase release rate, Annexin V positive cell rate and neutral red staining degranulation rate were significantly increased. CONCLUSION: Chlorogenic acid, as a small molecule substance, exists in the form of hapten, which itself does not have sensitization, but it can form immunogenicity by binding with serum protein and cause allergic reaction. Chlorogenic acid as Shuanghuanglian injection of an active ingredient, but also Shuanghuanglian injection caused by an allergy allergens.