大鼠皮肤类过敏试验的方法学研究及适用性考察

Methodological and Applicability Study of Rat Skin Anaphylactoid Test

目的研究大鼠皮肤类过敏试验方法的适用性和可靠性，对中药注射剂潜在的类过敏反应进行检测和评价。方法以C48／80为工具药物，对可能影响大鼠皮肤类过敏试验的灵敏度和可靠性因素进行比较研究，优选最佳试验条件，选用Tween80、内毒素、中华眼镜蛇毒素、天花粉蛋白注射液、痰热清注射液和血塞通注射液进行考察和应用。结果大鼠皮肤类过敏反应试验方法最佳条件为：采用胰岛素注射器（口径0．33mm）进行药物注射，50-100pL／点，给药后立即尾静脉注射0．5％伊文思蓝染液1mL，15min时颈动脉放血处死大鼠，剪取背部皮肤处理。采用该方法，Tween80、内毒素、中华眼镜蛇毒素、天花粉蛋白注射液均呈现一定的诱导大鼠皮肤出现蓝斑的能力，痰热清注射液给予临床剂量无蓝斑出现。血塞通注射液，临床剂量虽可引起大鼠皮肤蓝斑，但不排除与因其本身的药效作用干扰所致。结论该方法操作简便，试验周期短，需用药量少，检测敏感性高，结果重复性好，但易受药物本身特性干扰而出现假阳性。

Objective To study the sensitivity and reliability of rat skin anaphylactoid test method, detect and evaluate the anaphylactoid reaction of traditional Chinese medecine injection. Methods The condition of rat skin anaphylactoid test was optimized by studying the influencing factors of sensitivity and reliability of test with C48/80 as a tool drug, Tween80, endotoxin, China cobra toxin, trichosanthin injection, Tanreqing injection and Xuesaitong injection were investigated. Results The best conditions of rat skin anaphylactoid test was as follows： intrademal inject the drug with insulin syringe, 50-100μL per point, immediately inject 0.5% Evans blue dye 1 mL though caudal vein, 15 rain later, kill the rat by carotid artery bleeding, clip dorsal skin to do the test. With this method, Tween80, endotoxin, China cobra toxin and trichosanthin injection all can induce blue stain in rat skin. Tanreqing injection showed no blue stain at the clinical dose. Xuesaitong injection although can induce blue stain in rat skin at the clinical dose, but the results cannot exclude the interference of its pharmacological function. Conclusion The method is simple with short test cycle, less dose of test drug, high detection sensitivity and good reproducibility, but some drug can show false positive result due to its own property.