一种新的小鼠过敏试验模型的建立及其在中药注射剂致敏性评价中的应用  

A novel mouse allergy tested model and its application for traditional Chinese medicine injections′s allergy evaluation

作　　者：易艳[1] 李春英[1] 赵雍[1] 刘素彦[1] 韩佳寅[1] 张宇实[1] 梁爱华[1] YI Yan;LI Chun-ying;ZHAO Yong;LIU Su-yan;HAN Jia-yin;ZHANG Yu-shi;LIANG Ai-hua(State Key Laboratory of Innovative Natural Medicine and Traditional Chinese Medicine Injections,Key Laboratory of Beijing for Identification and Safety of Chinese Medicine,Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]中国中医科学院中药研究所中药鉴定与安全性检测评估北京市重点实验室创新天然药物与中药注射剂国家重点实验室,北京100700

出　　处：《中国中药杂志》2022年第13期3581-3588,共8页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金重点项目(82192913);北京市自然科学基金项目(7214291);中国中医科学院基金项目(C12021A04801,ZZ-13-035-10,GH201919)。

摘　　要：当药物诱导机体产生Ⅰ型过敏反应时,IgE与肥大细胞结合导致肥大细胞脱颗粒,释放血管活性物质,血管通透性增高,血管内物质可渗出血管外,该研究基于这一病理生理机制,建立一种可客观定量评价注射剂过敏反应的小鼠模型。将卵清蛋白(ovalbumin,OVA)腹腔注射致敏ICR小鼠,隔天1次,共致敏3次,末次致敏后14 d,致敏剂量4倍的OVA伊文思蓝(Evans blue,EB)混合溶液静脉激发,30 min后,根据致敏小鼠的耳廓蓝染面积和EB渗出量进行过敏反应强弱判断。结果与正常组比较,OVA 0.625/2.5、1.25/5、2.5/10、5/20 mg·kg^(-1)致敏/激发均能诱发小鼠发生明显以耳廓蓝染为主要表现的过敏反应;小鼠不致敏直接静脉注射OVA则不发生耳廓蓝染反应。上述OVA致敏小鼠血清进行小鼠被动皮肤过敏试验,在小鼠背部均可见明显蓝斑,并且5 mg·kg^(-1)致敏小鼠体内抗OVA-IgE含量明显升高,以上说明该小鼠模型可以检测OVA诱发的Ig介导的过敏反应。与诱发小鼠过敏反应相同剂量的OVA,同样也可诱发豚鼠发生较明显的过敏反应,提示该小鼠过敏反应模型敏感性也较高。另外,ICR和BALB/c小鼠与裸鼠相比,前两者对于OVA的致敏耳廓蓝染反应更敏感,OVA致敏ICR小鼠耳、肺组织显示明显的渗出炎症反应,血清中也检测出明显升高的炎性因子(VEGF和IL^(-1)0),提示该小鼠模型从病理上也与过敏反应具有一致性。选取的中药注射剂均不诱发小鼠和豚鼠发生明显Ⅰ型过敏反应,但有诱发小鼠类过敏反应风险。该模型能够很好地反映致敏原的致敏作用,并且操作简便、成本低、可重复性强,适用于IgE依赖的Ⅰ型过敏反应的预测和研究。When the drug induces the organism to produce a typeⅠallergic reaction,the combination of IgE and mast cells results in the degranulation of the mast cells.Release of vasoactive substances,increase in vascular permeability,and exudation of intravascular substances outside the blood vessels.Based on this pathophysiological mechanism,a mouse model that can objectively and quantitatively assess the allergic response to the injection has been established.ICR mice were sensitised by intraperitoneal injection of different doses of OVA once every two days for three times.14 days after the last sensitization,a combination OVA solution of 4 times the sensitizing dose and Evans blue were injected intravenously into mice for the challenge.Compared with the normal group,OVA 0.625/2.5,1.25/5,2.5/10,5/20 mg·kg^(-1) sensitized and challenged can induce allergic reactions mainly manifested by blue staining of the auricle in mice.Direct injection of OVA intravenously did not cause an auricular blue colouration reaction in mice.The passive cutaneous anaphylaxis reaction in mice was conducted with the aforementioned OVA-sensitized mouse serum,and there were obvious blue spots on the mouse′s back.In addition,the content of anti-OVA-IgE in 5 mg·kg^(-1)OVA-sensitized mice was significantly increased.Ears and lungs of mice sensitized to OVA showed evident exudation inflammation.Significantly elevated inflammatory factors(VEGF and IL^(-1)0)were also detected in the serum of OVA-sensitized mice.The equivalent dose of OVA caused obvious allergic reactions in both guinea pigs and mice.Compared with nude mice,ICR and BALB/c mice are more sensitive to OVA sensitization.Injections of selected TCMI did not induce typeⅠallergic reactions in mice and guinea pigs,but there was a risk of inducing pseu-doallergic reactions in mice.The model is problematic and may well reflect the sensitization effect of allergens.It obtains the benefits of simple operation,accuracy,low cost,easy extension,and high repeatability.It is suitable for predicting

关 键 词：动物模型 Ⅰ型过敏反应 血管通透性 炎症反应 中药注射剂 

分 类 号：R285.5[医药卫生—中药学]