清开灵、血塞通注射液类过敏反应研究

目的用RBL-2H3细胞初步评价清开灵、血塞通注射液类过敏反应,为完善中药注射剂类过敏反应的筛选提供实验依据。方法 MTT法测定2种药物对细胞生长的IC50;采用不同浓度药物、C48/80或培养液分别刺激细胞30 min,检测上清液中组胺、β-氨基己糖苷酶释放率;另取刺激后的细胞,观察其脱颗粒率及超微结构变化。取ICR小鼠进行类过敏试验,以含伊文思蓝的药物或生理盐水单次尾静脉注射30 min,记录每组耳廓蓝染的动物数、蓝染总耳数及耳廓伊文思蓝渗出量。结果离体实验显示,清开灵、血塞通注射液的IC50均为12.5μL/mL;2种药物在较高浓度均能促进细胞释放组胺和β-氨基己糖苷酶(P0.05,P0.01),呈一定剂量依赖性;形态上可见细胞表面绒毛减少,内部空泡状结构增加。小鼠类过敏试验显示,2种药物对动物耳蓝染率均为100%、伊文思蓝渗出量明显增多(P0.01)。离体与整体实验结果一致。结论清开灵、血塞通注射液具有潜在的致类过敏反应性,RBL-2H3细胞模型用于中药注射剂类过敏性评价具有一定的参考应用价值。

Studies on Anaphylactoid Reactions Induced by Traditional Chinese Medicine Injections of Qingkailing and Xuesaitong

Objective To study anaphylactoid reactions induced by traditional Chinese medicine injections （TCMI） of Qingkailing （QKL） and Xuesaitong （XST） with RBL-2H3 cells; To provide some reference for improving the screening system of TCMI induced anaphylactoid reactions. Methods The IC50 induced by QKL and XST injections was determined by MTT assay. RBL-2H3 cells were stimulated with TCMI at different concentrations or with C48/80 or culture medium. Thirty minutes later, the supernatant was collected to determine the release of histamine and β-hexosaminidase. The cell degranulation rate and the ultrastructure changes were observed. The ICR mice were given single injection of TCMI containing Evans Blue through tail vein. The number of the animal with blue ear, the total number of blue ears and the quantity of Evans Blue of extravasation were determined 30 minutes later. Results The IC50 of both QKL injection and XST injection was 12.5 ]aL/mL. These two injections promoted RBL-2H3 cells to release histamine and -hexosaminidase at higher concentrations （P 〈 0.05, P 〈 0.01） in a dose dependent manner. Cell morphology showed a decrease of villous on the cell surface and an increase of the internal vacuolated structure. Both injections caused the blue ears of all animal with a rate of 100%. The quantity of Evans Blue of the extravasation was significantly increased （P〈0.01）. The results in vitro study were in close agreement with that in vivo. Conclusion Both QKL injection and XST injection may potentially cause anaphylactoid reactions. The RBL-2H3 cell model may be valuable to evaluate the anaphylactoid reactions induced by TCMI.