基于RBL-2H3细胞的复杂基质生化药类过敏反应评估及体内外相关性评价  

Evaluation of anaphylactoid reactions induced by complex matrix biochemical drugs based on RBL-2H3 cells and its in vivo correlation

作　　者：吴畅 杨奇涛 王瑶 吴殷囡 刘洋 孟长虹 陆益红 汪玉馨 史清水 WU Chang;YANG Qi-tao;WANG Yao;WU Yin-nan;LIU Yang;MENG Chang-hong;LU Yi-hong;WANG Yu-xin;SHI Qing-shui(Xuzhou Medical University,Xuzhou Jiangsu 221004;Jiangsu Institute for Food and Drug Control,Nanjing 210019;China Pharmaceutical University,Nanjing 211198)

机构地区：[1]徐州医科大学,江苏徐州221004 [2]江苏省食品药品监督检验研究院,南京210019 [3]中国药科大学,南京211198

出　　处：《中南药学》2023年第8期1998-2003,共6页Central South Pharmacy

基　　金：江苏省药品监督管理局科研计划课题(编号:202119)。

摘　　要：目的 建立适宜的细胞模型对复杂基质生化药骨肽类注射剂和垂体后叶注射剂类过敏反应风险进行评估,并与类过敏反应体内评价模型实验结果的相关性进行比较。方法 采用RBL-2H3细胞脱颗粒实验,选择β-氨基己糖苷酶作为适宜的评价指标以评估骨肽类注射剂和垂体后叶注射剂对RBL-2H3细胞脱颗粒的影响;采用实验室已建立的类过敏反应体内评价模型小鼠耳郭蓝染实验,以蓝染率及伊文思蓝升高率为指标评价两种复杂基质生化药注射剂体内类过敏反应风险的相关性。结果 骨肽类注射剂可诱导β-氨基己糖苷酶的释放率增加,与阴性对照组相比差异存在统计学意义(P <0.01),垂体后叶注射剂不可诱导β-氨基己糖苷酶的释放。小鼠耳郭蓝染实验高剂量组中,复方骨肽注射液和骨肽注射液为临床等效剂量的6.67、10倍,类过敏反应阳性率分别为100%、66.7%;随着剂量的降低,复方骨肽注射液和骨肽注射液为临床等效剂量的1.67、2倍时,类过敏反应均判定为阴性,垂体后叶注射剂类过敏反应均为阴性,与体外结果一致。结论通过结合RBL-2H3细胞脱颗粒模型与小鼠耳郭蓝染模型可对药物类过敏反应风险进行评估和筛查,同时也可为复杂基质类生化药类过敏反应物质基础及机制的研究评价提供参考和手段。Objective To establish an appropriate cell model to evaluate the risk of anaphylactoid reactions induced by complex matrix biochemical drugs such as osteopeptide injections and posterior pituitary injections,and to compare the correlation with the results of evaluation model of anaphylactoid reactions in vivo.Methods RBL-2H3 cell degranulation test was done andβ-hexosaminidase(β-Hex)was used as an evaluation indicator to determine the effect of osteopeptide injections and posterior pituitary injections on RBL-2H3 cell degranulation.The mouse ear blue staining test was conducted in the laboratory before,which was used to evaluate the anaphylactoid reaction in vivo,and the risk of anaphylactoid reactions in vivo between two complex matrix biochemical injections with the increase rate in blue staining rate and Evans blue(EB)as indicators.Results Osteopeptide injections induced the increase release ofβ-Hex,with a statistical difference when compared with the negative control group(P<0.01).The posterior pituitary injections failed to induce the release ofβ-Hex.Among the high-dose groups on the mouse ear blue staining test,when the compound bone peptide injections and bone peptide injections were 6.67 and 10 time of the clinical equivalent dose,the positive rates of anaphylactoid reactions were 100%and 66.7%.However,with the decrease of dose,when the compound bone peptide injections and bone peptide injections were 1.67 and 2 times of the clinical equivalent dose,and the anaphylactoid reactions were negative.The anaphylactoid reactions of the posterior pituitary injection were negative,which was consistent with in vitro the results.Conclusion Combination of RBL-2H3 cell degranulation model and mouse ear blue staining model can assess and screen the risk of anaphylactoid reactions,and also provide reference and means for research and evaluation of the substance basis and mechanism of anaphylactoid reactions of complex matrix biochemical drugs.

关 键 词：类过敏反应 复杂基质生化药 RBL-2H3细胞 小鼠耳郭蓝染实验 

分 类 号：R392.8[医药卫生—免疫学]