寒证和热证动物模型的建立方法及评价  

Establishment and Evaluation of Animal Models with Han Syndrome and Re Syndrome

作　　者：李慧璇 吕智豪 覃景春 梁裕琪 文彬[1] Li Huixuan;Lii Zhihao;Qin Jingchun;Liang Yuqi;Wen Bin(Piwei Institute,Guangzhou University of Chinese Medicine,Guangzhou 510405;The First Clinical Medical College,Guangzhou University of Chinese Medicine,Guangzhou 510000)

机构地区：[1]广州中医药大学脾胃研究所,广州510405 [2]广州中医药大学第一临床医学院,广州510000

出　　处：《中药药理与临床》2020年第1期222-227,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(NO.81673944)。

摘　　要：目的:建立稳定、客观的中医寒证和热证的动物模型。方法:将72只Wistar大鼠(雌雄各半),随机分为正常对照组、寒证模型组和热证模型组。寒证模型组予冰水(0℃)灌胃和冷水(10℃)泡浴,热证模型组予乙醇溶液(0.1 mg/ml^0.3 mg/ml)和辣椒素(0.2 mg/ml^0.9 mg/ml)溶液灌胃,正常对照组予生理盐水灌胃。每组均予10 ml/kg灌胃,每日1次,连续5周。每日测定大鼠体征评分(R)、饮水量及摄食量,每周检测体重,取材前1 h测量大鼠肛温。造模结束后,测定大鼠大肠组织Na^+-K^+-ATP酶、Ca^2+-Mg^2+-ATP酶、琥珀酸脱氢酶(SDH)和乳酸脱氢酶(LDH)活性。HE染色,光镜下观察各组大鼠大肠组织学形态改变,并对病理组织学结果进行评分。结果:与正常对照组比较,寒证模型组6<R≤3时,评分显著高于正常对照组,饮水量和肛温明显降低,摄食量和体重增加。热证模型组R≥6时,评分显著高于正常对照组,饮水量和肛温上升,摄食量和体重下降。与正常对照组比较,热证模型组Ca^2+-Mg^2+-ATP酶、SDH酶和LDH酶活性明显升高;寒证模型组Na^+-K^+-ATP酶、Ca^2+-Mg^2+-ATP酶、SDH酶和LDH酶活性降低不明显。与热证模型组比较,寒证模型组Na^+-K^+-ATP酶、Ca^2+-Mg^2+-ATP酶、SDH酶和LDH酶活性显著降低(P<0.05或P<0.01)。病理组织学检查表明,寒证模型、热证模型组大肠粘膜杯状细胞排列紊乱,炎性细胞浸润。病理评分结果表明,寒证模型、热证模型组病理损伤明显高于正常对照组(P<0.05或P<0.01)。结论:寒证模型通过冰水灌胃和冷水泡浴的方法建立,热证模型通过乙醇溶液和辣椒素溶液灌胃建立,模型成功模拟寒证、热证的临床症状及病理组织学变化。本实验结果为寒证、热证模型的病理生理及药理研究提供了更为准确、客观、有效的动物模型评价方法。Objective: To establish a stable and objective animal model with Han syndrome and Re syndrome. Methods: 72 Wistar rats( male and female) were randomly divided into the control group,Han model group and Re model group. Rats in the Han model group were orally administered with ice water( 0 ℃) and bathed in cold water( 10 ℃) . Rats in the Re model group were administered with a solution of ethanol solution( 10 % to 30 %) and capsaicin( 0. 2 ~ 0. 9 mg/ml). Rats in the control group were orally given normal saline. Rats in each group were given 10 ml/kg once a day for 5 weeks. The rat’s physical score( R),water intake and food intake were measured daily,the body weight was measured every week,the anal temperature of rat was measured 1 h before samples were taken. After modeling,the activities of Na^+K^+-ATPase,Ca^2+Mg^2+-ATPase,succinate dehydrogenase( SDH),and lactate dehydrogenase( LDH) in large intestine tissues were measured.After HE staining,the histological changes of the large intestine in each group were observed under light microscope,and the histopathological results were scored. Results: Compared with the control group,when 6 < R≤3 was observed in Han model group,the score was significantly higher than that in the control group,the water consumption and anal temperature were significantly reduced,the food intake and body weight were increased. When R ≥ 6 was observed in Re model group,the score was significantly higher than that in the control group,the water intake and anal temperature were increased,the food intake and body weight were decreased. Compared with the control group,the activities of Ca^2+Mg^2+-ATPase,SDH enzyme and LDH enzyme in Re model group were significantly increased. The activities of Na^+K^+-ATPase,Ca^2+Mg^2+-ATPase,SDH enzyme and LDH enzyme in Han model group were not significantly reduced. Compared with the Re model group,the activities of the Na^+K^+-ATPase,Ca^2+Mg^2+-ATPase,SDH enzyme,and LDH enzyme in Han model group were significantly reduced. Histopathological examinat

关 键 词：寒证 热证 动物模型 腺苷三磷酸酶(ATP) 乳酸脱氢酶(LDH) 琥珀酸脱氢酶(SDH) 

分 类 号：R-332[医药卫生]