中药注射剂的类过敏实验动物模型和实验方法研究

目的：探讨中药注射剂类过敏实验动物模型和实验方法，以用于中药注射剂类过敏反应的临床前评价。方法：（1）Beagle犬随机分成：0.5% T-80组、鱼腥草蒸馏液组、含0.5%吐温的鱼腥草注射液组、0.5% HS15组、含0.5% HS15的鱼腥草注射液组。上述各组分别静脉注射给予各自受试药3mL·kg-1。给药后观察动物的反应，给药前和给药后10、30min分别测定血压、呼吸、心率。（2）小鼠或豚鼠分别一次性静脉注射给予不同浓度 T-80溶液、含不同浓度 T-80的鱼腥草蒸馏液、不含 T-80的鱼腥草蒸馏液，Compound 48/80溶液；以上受试物中均含有0.4%伊文思蓝。记录给药后各组动物耳廓血管通透性增高反应的阳性率，按照耳廓蓝染程度评分，并采用分光光度计定量测定耳廓伊文思蓝渗出量。结果：含T-80的受试物（0.5% T-80组单独或含0.5% T-80的鱼腥草注射液）给Beagle犬一次性静脉注射后，犬迅速出现类过敏反应，包括面部、口唇、眼结膜和耳部皮肤黏膜红肿、瘙痒，流涎、流泪，步态不稳或跌倒，精神萎靡或嗜睡，呼吸急促，血压下降等。在小鼠和豚鼠上，不同浓度的T-80或含不同浓度T-80的鱼腥草注射液可剂量依赖性地导致耳廓血管通透性增高。而其它不含T-80的受试物给Beagle犬静脉注射后未见类过敏反应，给小鼠和豚鼠注射后未见耳廓血管通透性增高反应。结论：鱼腥草注射液可导致类过敏反应，T-80是其主要的致类过敏反应物质。Beagle犬、小鼠和豚鼠均可作为中药注射剂类过敏实验的动物模型。Beagle犬的类过敏反应与临床高度一致，模型敏感性高、重复性好。小鼠和豚鼠的类过敏反应特征主要表现为耳廓血管通透性增高，该血管反应符合类过敏反应发生机制。

Animal Models and Methodologies for Evaluation of Chinese Herbal Injection-induced Pseudoanaphylactoid Reactions

This study aimed at development of animal models and methodologies for preclinical evaluation of pseudoanaphylactoid reactions induced by Chinese herbal injections. Beagle dogs were divided into the group of 0.5% Tween 80, distilled Yuxingcao solution, Yuxingcao injection containing 0.5% Tween 80, 0.5% HS 15, Yuxincao injection containing 0.5% HS 15. Different groups of Beagle dogs were given 3ml/kg of the intravenous injection respectively. Anaphylactoid reactions were observed. The blood pressure, respiratory rate and heart rate were tested before the injection, 10 min and 30 min after the injection. Mice or guinea pigs were intravenously injected solutions with different concentrations of Tween 80, Yuxingcao injection containing different concentrations of Tween 80, distilled Yuxingcao solution, or positive control compound 48/80. All test substances were mixed with 0.4% Evans blue. Reactions and vascular permeability of the ear was observed and measured 30 min after injection. The results showed that a variety of anaphylactoid reactions, such as cutaneous and mucosa signs of the lip, conjunctiva, ear and circumoral skin, bronchospasma, cardiovascular collapse, somnolence, lethargy, breathless or dyspnea, severe hypotension, were observed immediately after the injection of Tween 80 and Yuxingcao injection containing 0.5% Tween 80 among Beagle dogs. Those reactions occurred at both first injection or repeated injections at intervals of 2 weeks or 4 weeks, manifesting that it was pseudoanaphylactoid reaction mediated by non-immune mechanisms. In mice and guinea pigs, single dose of intravenous injection of solutions with various concentrations of Tween 80 or Yuxingcao injection containing different concentrations of Tween 80 caused vascular hyperpermeability in the ear. Yuxingcao injection causes pseudoanaphylactoid reactions and its solubilizer Tween 80 is the principal contributor for pseudoanaphylactoid reactions. Beagle dogs, mice and guinea pigs can be used as animal model for preclinical evaluation of pseudoanaphylactoid reactions for Chinese herbal injections. Pseudoanaphylactoid reactions in Beagle dogs are of high clinic consistency, sensitivity, reproducibility, and maneuverability. Vascular hyperpermeability which represents the basic mechanism of pseudoanaphylactoid reactions is the main characteristic reactions in mice and guinea pigs.