构建被动皮肤过敏反应大鼠模型并检测其肠系膜淋巴动力学变化

Passive cutaneous anaphylaxis reaction rats model and their changes of mesentery lymph dynamics

背景:Ⅰ型变态反应性疾病的常用动物模型是被动皮肤过敏反应(passive cutaneous anaphylaxis reaction,PCA),目前对PCA发病的淋巴微循环机制尚不清楚。目的:构建PCA大鼠模型,并观察其肠系膜淋巴动力学变化。设计、时间及地点:随机对照动物实验,于2006-12/2007-03在河北北方学院病理生理实验室完成。材料:清洁级雄性Wistar大鼠58只,其中6只用于抗血清制备,20只用于蓝色反应斑测定,32只用于肠系膜淋巴微循环观察,用于蓝色反应斑测定和肠系膜淋巴微循环观察的大鼠均随机分为4组,对照组、PCA1组、PCA2组、PCA3组。方法:用卵清蛋白作抗原制备抗血清,分别于PCA1,PCA2,PCA3组大鼠背部脊柱两侧去毛处皮内注射5%,10%,20%抗血清,48h后再用10%卵清蛋白攻击致敏,复制PCA模型;对照组用生理盐水代替。主要观察指标:①蓝色反应斑测定判断模型是否成功。②肠淋巴管插管后引流卵清蛋白攻击前10min至攻击后30min不同时间段的淋巴液,计算淋巴流出量、细胞总数计数、单核细胞分类计数、淋巴细胞输出量。结果:PCA1,PCA2,PCA3组蓝色反应斑A值均显著高于对照组(P<0.05)。卵清蛋白攻击10min以后,PCA1,PCA2,PCA3组的肠淋巴流量、细胞总数、单核细胞数、淋巴细胞输出量均显著高于对照组及攻击前水平(P<0.05)。结论:①成功复制了大鼠PCA模型,此模型可反映Ⅰ型变态反应性皮肤病时的病理生理改变,并有良好的可操作性和可重复性。②PCA发生后,淋巴回流动力及淋巴转运功能明显增强,可能是导致变态反应性皮肤病皮损区淋巴细胞浸润的重要因素之一。

BACKGROUND： Animal model of passive cutaneous anaphylaxis （PCA） reaction is very common for type I allergic reaction dermatosis, but lymph microcirculation of PCA onset is absent at present. OBJECTIVE： To develop the PCA model in rats, and observe the changes of mesentery lymph dynamics on PCA reaction rats. DESIGN, TIME AND SETTING： A randomized control animal experiment was carried out in the Laboratory of Pathophysiology, Hebei North University （Zhangjiakou, Hebei Province, China） between December 2006 and March 2007. MATERIALS： Fifty-eight male Wistar rats of clean grade involved in this study, 6 of them were used to prepare antiserum, 20 ones were used for the determination of blue plaque, and 32 ones were used to observe the lymph microcirculation. Rats were randomly divided into 4 groups： control group, PCA1 group, PCA2 group and PCA3 group. METHODS： Ovalbumin （OVA） was taken as antigen to prepare antiserum, the model of PCA was established by removing their back hair and administrating the prepared 5%, 10%, and 20% antiserum on two sides with endermic injection in PCA1, PCA2, and PCA3 groups. Then administration of 10% OVA lead to hypersensitivity 48 hours later, while normal saline was replaced in the control group. MAIN OUTCOME MEASURES： Blue plaque determination was used to verify the PCA model. For the rats in each group, the lymph was taken out after intestine lymphatic intubatton from 10 minutes before OVA attack to 30 minutes after OVA attack. The lymph flow, total cell number, monocyte number, and lymphocyte output from intestinal lymph duct were observed at all periods. RESULTS： Results of blue plaque determination showed that, the absorbance value of PCA1, PCA2 and PCA3 groups was significantly higher than that in the control group （P 〈 0.05）. After 10 minutes of OVA attack, the lymph flow and total cell number, monocyte number, lymphocyte output from intestinal lymph duct in PCA1, PCA2 and PCA3 groups were significantly increased than that in the control group and before OVA attack （P 〈 0.05）. CONCLUSION： The PCA model has been successfully duplicated in rats, and this model can reflect the pathophysiology changes of allergic reaction dermatosis of type I, with the well operability and repetitiveness. Both lymphatic return dynamia and lymph transport function increase remarkably after PCA, which is one of the important factors to result in lymphocyte infiltration in skin lesion area of allergic dermatogic disease.