【作者】 肖俊霞；

【导师】 尹卫东； 周秀田；

【作者基本信息】 南华大学 ， 病理学与病理生理学， 2005， 硕士

【摘要】 目的:观察高脂/高糖/高胆固醇饲料能否引起广西巴马小型猪发生糖脂代谢紊乱,从而发展为2型糖尿病;观察NO-1886能否改善糖脂代谢紊乱,保护胰岛β细胞;观察NO-1886对胰岛β细胞凋亡的影响及其对细胞凋亡相关基因蛋白Bcl-2和Bax表达的影响,从而阐明这一新的药物对胰腺β细胞凋亡的影响和β细胞凋亡在2型糖尿病的发病机制中的作用。 方法:1) 动物及喂养:广西巴马猪随机分为对照组、高脂/高糖/高胆固醇喂养组、高脂/高糖/高胆固醇喂养+NO-1886治疗组;分别用正常饲料、高脂/高蔗糖/高胆固醇饲料、高脂/高蔗糖/高胆固醇饲料补充1.0%NO-1886喂养5个月。2)血糖、血脂代谢研究等:每个月末取动物空腹血,测定血糖、胰岛素、胰高血糖素、TG、TC、Amylin、FFA、HDL-C。每个月末作一次口服葡萄糖耐量试验。3) 用光镜、电镜、胰岛素免疫组化染色、刚果红染色、Amylin免疫组化、Hoechest荧光染色观察胰岛细胞凋亡;胰岛素免疫组化+细胞凋亡原位检测(TUNEL)标记β细胞凋亡计数。4) 用免疫组化和Western blot技术检测bcl-2、bax在胰岛的表达。 结果:用高脂/高蔗糖/高胆固醇饲料喂养可诱导广西巴马猪发生糖代谢的紊乱,具有2型糖尿病的特征,引起空腹葡萄糖、胰岛素、胰高血糖素、TG、TC、FFA、Amylin水平增高,HDL-C水平降低以及胰岛素抵抗和胰岛β细胞损害,胰岛内有IAPP的沉积,胰岛bcl-2蛋白表达下调、bax蛋白表达上调。在高脂高蔗糖高胆固醇饲料中添加1%NO-1886后,巴马猪空腹血糖、胰岛素、胰高血糖素、TG、TC、FFA、Amylin浓度均无明显升高,HDL-C水平增高,胰岛内IAPP沉积减少,血浆葡萄糖清除率以及胰岛素分泌急性相都有了明显改善,胰岛β细胞的结构和功能受到保护。胰岛bcl-2蛋白表达上调、bax蛋白表达下调。更多还原

【Abstract】 Objectives To observe high-fat/high-sucrose/high-cholesterol diet inducing the changes of glucose and lipid metabolism and try to establish a new animal model of the human type 2 diabetes mellitus; to investigate the effects of NO-1886 on glucose and lipid metabolism and functions of the pancreatic islet; To study the effects of NO-1886 on β-cell apoptosis and the protein expression of bcl-2/bax from the islet in Guangxi Bama minipigs.Materials and methods Animals and diets: Guangxi Bama minipigs were randomized into three groups with similar body weight: control group, high-fat /high-sucrose/high-cholesterol group, high-fat/high-sucrose/high-cholesterol with NO-1886 treated group. The animals were fed with control diet, high-fat/high-sucrose/ high-cholesterol diet and high-fat/high-sucrose/high-cholesterol diet supplemented with 1%NO-1886 for 5 months. Blood samples were withdrawn from the orbital sinus of the animals at the end of each month after fasting overnight. Blood glucose, insulin, glucagons, TG, TC, Amylin, FFA and HDL-C were determined. An oral glucose tolerance test (OGTT) was performed every month. The animals were sacrificed at the end of month 5. The pancreas tissues were dissected from each animal. The effects of NO-1886 on morphology of pancreas were analysed by HE stain, immuno-histological stain, fluorescence staining, Congo Red stain, the apoptosis of p-cell was observed by light and electron microscopy. The levels of bcl-2 and bax in islet were measured by immunohistochemistry and Western blot.Results Feeding a high-fat/high-sucrose/high-cholesterol diet to miniature pigs increased plasma levels of TG, TC, FFA, glucose, glucagon, Amylin, and decreased the更多还原