清开灵注射液类过敏反应的实验研究

Experimental study on pseudoallergic reaction of Qingkailing injection

实验采用小鼠全身类过敏试验、大鼠皮肤类过敏试验及测量血液中组胺,VEGF,TNF-α含量来研究清开灵注射液致小鼠和大鼠的类过敏反应,为其临床用药安全提供参考。小鼠全身类过敏试验:ICR小鼠随机分为生理盐水(NS)对照组、阳性药Compound 48/80(Comp)25 mg·kg-1组、清开灵注射液(QKLI)3,6,12 mL·kg-1组(本实验所用QKLI为液体,故剂量单位选用mL·kg-1),各组分别一次性尾静脉注射含0.4%伊文思蓝(EB)的受试药,记录给药30 min后耳廓蓝染阳性率,对耳廓蓝染程度进行评分,测定耳廓伊文思蓝渗出量。大鼠皮肤类过敏试验:剃除大鼠背部和脊柱两侧的毛,经尾静脉注射含0.6%EB的NS 2 mL·kg-1,10 min后,在麻醉的情况下,沿脊柱两侧,皮内注射阴性对照NS、阳性对照Comp(0.5 mg,50μL)、不同浓度QKLI(分别相当于清开灵注射液原液12.5,25,50μL),各组注射体积均为50μL。给药20 min后记录各注射点的蓝斑直径。对血液中组胺,VEGF,TNF-α的影响:ICR小鼠随机分成NS组、Comp 25 mg·kg-1组、QKLI 12 mL·kg-1组。各组经尾静脉注射不含EB的受试物。给药30 min后,摘眼球取血,离心分离血清,采用ELISA方法测定。实验结果显示尾静脉注射QKLI 3,6,12 mL·kg-1后,小鼠耳廓出现不同程度的耳蓝染现象。提示QKLI可引起小鼠血管通透性增高,导致其发生类过敏反应;大鼠皮内注射QKLI(相当于原液25,50μL/点)时可导致注射点处形成与NS组相比直径较大的蓝斑(P<0.01);静脉注射QKLI后可引起血液VEGF明显增高,组胺有一定增高趋势,TNF-α未见增高。实验证明了QKLI有显著的使血管通透性增高的作用,从而诱发大鼠、小鼠而发生类过敏反应,其类过敏反应可能与组胺和VEGF释放有关。

Pseudoallergic reactions of Qingkailing injection （QKLI） was assessed by vascular hyperpermeability which were indicated by ear blue staining in ICR mice after single intravenous injection of QKLI mixed with Evans blue （EB） and skin blue spot formation in SD rats after intradermal injection of QKLI and intravenous injection of EB. In addition, QKLI-induced histamine, VEGF, TNF-a release was measured after ICR mice received the single dosing of QKLI iv. The mild vascular hyperpermeability characterized by ear blue staining could be observed in mice after intravenous injection of QKLI and EB. Intracutaneous injection of 50 uL of test solution containing QKLI （25,50 uL） in rat back skin caused obvious local vascular hyperpermeability at the injection sites so as to result the larger diameters of blue spots than that in negative control group （ P 〈 0. 01 ）. QKLI induced a significant increase of VEGF and a slight elevation of histamine in mice after intravenous administration, while TNF-a showed no change after QKLI iv. The results in this study indicated that both intravenous injection and intracutanous injection of QKLI could induce vascular hyperpemeability so as to cause pseudoallergic reaction in mice and rats. QKLI-induced pseudoallergic reaction may be associated with the release of histamine and VEGF.