目的 建立中藥注射劑體外類過敏反應(yīng)評價方法，快速評價不同批次注射用益氣復(fù)脈（凍干）類過敏反應(yīng)表現(xiàn)。方法 體外培養(yǎng)嗜堿性白血病細(xì)胞株RBL-2H3細(xì)胞，選擇Compound 48/80為陽性藥，采用實時細(xì)胞分析（real-time cell analysis，RTCA）系統(tǒng)檢測藥物干預(yù)后引起的細(xì)胞指數(shù)（CI）值變化，并利用甲苯胺藍(lán)、鬼筆環(huán)肽染色觀察細(xì)胞形態(tài)、骨架變化，以及檢測組胺和β-已糖苷酶釋放量驗證RBL-2H3細(xì)胞的脫顆粒情況。選擇20個批次的注射用益氣復(fù)脈（凍干，100 μg/mL）作用于RBL-2H3細(xì)胞，進(jìn)行基于RTCA技術(shù)的類過敏反應(yīng)評價。結(jié)果 陽性藥Compound 48/80（20 μg/mL）能夠使RBL-2H3細(xì)胞的CI值在加藥后30 min內(nèi)呈先快速上升后下降趨勢；形態(tài)學(xué)研究發(fā)現(xiàn)，Compound 48/80使細(xì)胞形態(tài)和細(xì)胞骨架均發(fā)生明顯改變，發(fā)生明顯的脫顆?，F(xiàn)象；組胺和β-己糖苷酶釋放實驗進(jìn)一步證實Compound 48/80導(dǎo)致炎癥介質(zhì)的釋放，引起了明顯的脫顆?，F(xiàn)象；提示RTCA系統(tǒng)可以用于快速敏感的評價RBL-2H3細(xì)胞脫顆粒。不同批次的注射用益氣復(fù)脈（凍干）對RBL-2H3細(xì)胞CI值無明顯影響，提示所選批次為合格批次，無類過敏反應(yīng)現(xiàn)象的發(fā)生。結(jié)論 建立了一套基于RTCA系統(tǒng)的類過敏反應(yīng)體外快速評價技術(shù)，可用于注射用益氣復(fù)脈（凍干）等中藥注射劑類過敏反應(yīng)的體外快速評價。

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.