Pseudoallergy is a typical and common adverse drug reaction to injections, especially in traditional Chinese medicine injections (TCMIs). At present, the evaluation methods for pseudoallergy include cell methods in vitro and animal methods in vivo. The mast cell evaluation method based on the β-hexosaminidase (β-Hex)-catalyzed substrate, 4-nitrophenyl-β-N-acetyl-D-glucosaminide (4-NPG), is an important method for the evaluation of drug-induced pseudoallergy, but it is prone to false positive results and has insufficient sensitivity. In this study, a novel β-Hex evaluation system with rat basophilic leukemia-2H3 cells based on high-performance liquid chromatography-fluorescence detection (HPLC-FLD) was established, which effectively increased the sensitivity and avoided false positive results. Cell viabilities were measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide assay. In addition, a method for the determination of histamine, which is another indicator in the development of pseudoallergy, was established to validate the above method. The results of this novel method indicated that two TCMIs (Shuxuening injection and Shenqi Fuzheng injection), which were considered to be pseudoallergenic using 4-NPG, were not pseudoallergenic. Overall, the novel β-Hex/HPLC-FLD evaluation system using Rat basophilic leukemia-2H3 cells established was effective and precise. It could be used for the evaluation of pseudoallergic reactions caused by TCMIs and other injections.

中文翻译：

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基于β-己糖胺酶的假性过敏评价新方法及其在中药注射剂中的应用

假性过敏是注射剂特别是中药注射剂中典型且常见的不良反应。目前，假性过敏的评价方法包括体外细胞法和体内动物法。基于β-己糖胺酶(β-Hex)催化底物4-硝基苯基-β-N-乙酰基-D-氨基葡萄糖(4-NPG)的肥大细胞评价方法是评价药物诱导的肥大细胞评价的重要方法。假性过敏，但容易出现假阳性结果，灵敏度不够。本研究建立了一种基于高效液相色谱-荧光检测（HPLC-FLD）的大鼠嗜碱性白血病-2H3细胞β-Hex评价体系，有效提高了灵敏度，避免了假阳性结果。通过3-(4,5-二甲基-2-噻唑基)-2,5-二苯基溴化四唑测定法测量细胞活力。此外，还建立了假性过敏发生的另一个指标——组胺的测定方法，以验证上述方法。该新方法的结果表明，使用4-NPG被认为具有假变应原性的两种中药制剂（舒血宁注射液和参芪扶正注射液）并非假变应原性。总体而言，建立的基于大鼠嗜碱性白血病-2H3细胞的β-Hex/HPLC-FLD评价体系是有效且精确的。可用于评价中药注射剂及其他注射剂引起的假性过敏反应。