目的 建立中药注射剂体外类过敏反应评价方法，快速评价不同批次注射用益气复脉（冻干）类过敏反应表现。方法 体外培养嗜碱性白血病细胞株RBL-2H3细胞，选择Compound 48/80为阳性药，采用实时细胞分析（real-time cell analysis，RTCA）系统检测药物干预后引起的细胞指数（CI）值变化，并利用甲苯胺蓝、鬼笔环肽染色观察细胞形态、骨架变化，以及检测组胺和β-已糖苷酶释放量验证RBL-2H3细胞的脱颗粒情况。选择20个批次的注射用益气复脉（冻干，100 μg/mL）作用于RBL-2H3细胞，进行基于RTCA技术的类过敏反应评价。结果 阳性药Compound 48/80（20 μg/mL）能够使RBL-2H3细胞的CI值在加药后30 min内呈先快速上升后下降趋势；形态学研究发现，Compound 48/80使细胞形态和细胞骨架均发生明显改变，发生明显的脱颗粒现象；组胺和β-己糖苷酶释放实验进一步证实Compound 48/80导致炎症介质的释放，引起了明显的脱颗粒现象；提示RTCA系统可以用于快速敏感的评价RBL-2H3细胞脱颗粒。不同批次的注射用益气复脉（冻干）对RBL-2H3细胞CI值无明显影响，提示所选批次为合格批次，无类过敏反应现象的发生。结论 建立了一套基于RTCA系统的类过敏反应体外快速评价技术，可用于注射用益气复脉（冻干）等中药注射剂类过敏反应的体外快速评价。
Objective To establish a new method for evaluating the anaphylactoid reaction of traditional Chinese medicine injection in vitro. Rapid evaluation of anaphylactoid reaction of Yiqi Fumai Lyophilized Injection (YQFM) from different batches was conducted. Method The basophilic leukemia cell line RBL-2H3 cells were cultivated in vitro, and Compound 48/80 was selected as the positive drug. The effect of Compound 48/80 on cell index (CI) was monitored by using real-time cell analyzer (RTCA). Then the cell morphology and cytoskeleton changes were observed by using toluidine blue and phalloidin staining, histamine and beta hexose glucosidase release were used to verify the degranulation of RBL-2H3 cells. Finally, 20 batches of YQFM (100 μg/mL) were selected for the anaphylaxis evaluation. Results Compound 48/80 could significantly increase the CI value of RBL-2H3 cells, further morphological study found that Compound 48/80 could make the cell morphology and cytoskeleton change obviously and show obvious degranulation at the same dose. Histamine and beta hexose glucosidase release experiments also confirmed that Compound 48/80 could lead to the release of inflammatory mediators and induce obvious degranulation at this dose. These results suggest that the RTCA system can be used to evaluate the degranulation of RBL-2H3 cells quickly and sensitively. RTCA monitoring results showed that different batches of YQFM had no significant effect on the CI value of RBL-2H3 cells, suggesting that the selected batches of YQFM were eligible batches without anaphylactoid reaction. Conclusion An in vitro rapid evaluation method for anaphylactoid reaction has been established based on RTCA system, which can be used for rapid evaluating the anaphylactoid reaction of YQFM and other traditional Chinese medicine injections in vitro.