大鼠动脉硬化闭塞模型两种构建方法的比较

背景：由于动脉硬化闭塞症的发病机制还不明确，所以建立动脉硬化闭塞症的病理动物模型，对于探明动脉硬化闭塞症的病因、病理生理、发病机制及防治药物的研究与开发均具有重要的意义。 目的：比较单纯高脂饲料喂养及给予内膜损伤加高脂饲料喂养大鼠动脉硬化闭塞模型复制建立的方法。 设计、时间及地点：随机对照动物实验，2007-08/2008-03于中国医科大学实验动物中心完成。 材料：清洁级3.0~4.0月龄健康纯种Wistar雄性大白鼠60只，体质量200 g。高脂饲料:62.8%基础饲料+20%猪油+150 g/L胆固醇+20 g/L胆酸钠+2 g/L丙基硫氧嘧啶。 方法：将60只大鼠随机分成3组，每组20只。普通饲养组给予普通饲料喂养；高脂饲养组给予高脂饲料喂养；高脂饲养加内膜损伤组给予内膜损伤加高脂饲料喂养。后两组同时给予维生素D3，30万u/kg体质量，右后肢肌肉注射,1次/月。 主要观察指标：①于内膜损伤后术后30 d，各组大鼠取血检测血浆总胆固醇、三酰甘油、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇浓度变化。②于内膜损伤后术后7，30，90 d时，取大鼠左后肢股-腘动脉，行苏木精-伊红等染色，光镜下观察股-腘动脉的病理变化。 结果：高脂饮食组大鼠与正常饮食组大鼠相比，高脂饮食组大鼠血清胆固醇 ，三酰甘油和低密度脂蛋白胆固醇均明显升高。病理显示：7 d高脂饲养组内皮细胞少量脱落，高脂饲养加内膜损伤组内皮细胞完全脱落、弹力板松弛，平滑肌细胞排列紊乱。90 d高脂饲养组内皮细胞完全脱落、平滑肌和胶原纤维增多、增厚，为早期硬化改变。高脂饲养加内膜损伤组内膜增厚、大量吞噬脂质、类脂质细胞、管腔闭塞。 结论：单纯给予大鼠高脂、高胆固醇饲料喂养不易形成动脉硬化闭塞症病变，大鼠股-腘动脉内注入蒸馏水损伤内皮加高脂饲料喂养可较快形成与人动脉硬化闭塞症相似的、较成熟的大鼠动脉硬化闭塞模型。

Comparison of constructing arteriosclerosis obliterans models with two methods in rats

BACKGROUND: The nosogenesis of arteriosclerosis obliterans (ASO) remains unclear. Therefore, construction ASO model has great significance for exploring the mechanism of etiology, pathogenesis of ASO and searching for preventive drugs. OBJECTIVE: To compare the constructing ASO models with two methods in rats DESIGN, TIME AND SETTING: A randomized control experiment of animals was performed at the Experiment Animal Center of China Medical University from August 2007 to march 2008. MATERIALS: Sixty healthy, male, 3.0-4.0 months old, clean grade Wistar rats, weighting 200 g, were selected. The high-fat diet was consist of 62.8% basic diet, 20% lard, 150 g/L cholesterol, 20 g/L sodium cholate, and 2 g/L propylthiouracil. METHODS: Rats were randomly divided into three groups, with 20 animals in each group. Rats in the basic diet and high-fat diet groups were fed normal and high-fat diet respectively. Rats in the high-fat diet plus intimal injury group were fed high-fat diet after intimal injury. All animals were intramuscular injected 3 000 000 u/kg vitamin D3 once per month except the normal diet group. MAIN OUTCOME MEASURES: The concentration change of total cholesterol, triglyceride, high-density lipoprotein cholesterol (HDL-C) and low-density cholesterol cholesterol (LDL-C) were measured at 30 days after operation. At 7, 30, 90 days after the intimal injury, the femoropopliteal artery of the left hind limb was removed and the pathological changes were observed by haematoxylin-eosin staining and light microscope. RESULTS: The serum total cholesterol, triglycerides, and low-density lipoprotein cholesterol in the high-fat diet group were significantly increased compared to the basic diet group. The pathological examination showed that at 7 days, endothelial cells in the high-fat diet group were exfoliated a little. While endothelial cells completely exfoliated, elastic plate relaxed, and smooth muscle cells arranged disordered in the high-fat diet plus intimal injury group. At 90 days, early changes of sclerosis appeared, including the endothelial cells completely exfoliated, increased smooth muscles, and collagen fibers thickening and hardening in the high-fat diet group. In the high-fat diet plus intimal injury group, the intimal was thickened, phagocytosed a large amount of lipid and lipid-like cells, with obliteration. CONCLUSION: Feeding rats with simple high-fat diets can not establish the ASO models easily, but, combining the high-fat diet with injected distilled water into rats femoropopliteal artery can induce a mature model, which is more similar to human ASO.