糖尿病心肌病巴马小型猪模型的制备  

Preparation of Bama miniature pig model of diabetic cardiomyopathy

作　　者：姚爽 沈艳明 于萍[1] 江仁美[1] 王彩梅[2] 邹迪莎 唐敏娟[1] 刘晓玲[1] 莫如芬[1] 施佳成 黄倩 于健[1] Yao Shuang;Shen Yanming;Yu Ping;Jiang Renmei;Wang Caimei;Zou Disha;Tang Minjuan;Liu Xiaoling;Mo Rufen;Shi Jiacheng;Huang Qian;Yu Jian(Department of Endocrinology,Affiliated Hospital of Guilin Medical College,Guangxi Zhuang Autonomous Region,Guilin 541001,Guangxi Zhuang Autonomous Region,China;Department of Clinical Laboratory,Affiliated Hospital of Guilin Medical College,Guang xi Zhuang Autonom ous Region,Guilin 541001,Guangxi Zhuang Autonomous Region,China;Department of Nephrology,Affiliated Hospital of Guilin Medical College,Guangxi Zhuang Autonomous Region,Gu ilin 541001,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西壮族自治区桂林医学院附属医院内分泌科,广西壮族自治区桂林541001 [2]广西壮族自治区桂林医学院附属医院检验科,广西壮族自治区桂林541001 [3]广西壮族自治区桂林医学院附属医院肾内科,广西壮族自治区桂林541001

出　　处：《中国中西医结合急救杂志》2019年第5期573-576,共4页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：广西壮族自治区科技计划项目(桂科AD16450020);国家临床重点专科建设项目〔桂医科(2018)5号〕;中青年教职工科研能力提升项目(2018glmcy056)。

摘　　要：目的观察糖尿病心肌病(DCM)巴马小型猪模型心肌组织核转录因子-κB p65(NF-κB p65)、转化长因子-β1(TGF-β1)及凋亡相关因子Bcl-2、Bax蛋白表达水平的变化及意义.方法选取4~5个月龄健康雄性广西巴马小型猪10头,按随机数字表法将动物分为对照组和模型组,每组5头.两组均空腹12 h后,采用一次性耳缘静脉注射链脲佐菌素(STZ)150 mg/kg的方法复制DCM巴马小型猪模型;对照组一次性耳缘静脉注射柠檬酸-柠檬酸钠缓冲液150 mg/kg.制模10个月后观察两组动物基本情况,并测定其空腹血糖(FPG)水平;采用蛋白质免疫印迹试验(Western Blot)检测两组心肌组织NF-κB p65、TGF-β1、Bcl-2、Bax的蛋白表达水平;电镜下观察两组动物心肌组织病理学变化.结果4头动物制模成功,1头死亡,模型成功动物出现多饮、多食、多尿及体质量下降等糖尿病表现.实验结束时模型组FPG水平明显高于对照组(mmol/L :25.53±3.75比4.68±0.77,P<0.01).与对照组比较,模型组动物心肌组织NF-κB p65、Bax、TGF-β1的蛋白表达水平均明显升高(NF-κB p65/GAPDH :0.46±0.05比0.38±0.02,Bax/GAPDH :0.46±0.01比0.35±0.01, TGF-β1/GAPDH :0.39±0.01比0.33±0.01,均P<0.05),Bcl-2蛋白表达水平明显降低(Bcl-2/GAPDH :0.33± 0.01比0.42±0.01,P<0.01).电镜下可见:模型组心肌组织肌原纤维排列较紊乱、间隙线粒体数量明显减少,可见大量空泡变性线粒体.结论采用一次性耳缘静脉注射大剂量STZ 10个月后可成功复制DCM巴马小型猪模型;DCM小型猪模型存在明显糖代谢紊乱,且心肌组织存在炎症反应、心肌细胞凋亡和纤维化表现.Objective To observe the changes and significance of the protein expression levels of nuclear factor-κB p65(NF-κB p65),transforming long factor-β1(TGF-β1)and apoptosis-related factors Bcl-2 and Bax in myocardial tissue of Bama miniature pig model of diabetic cardiomyopathy(DCM).Methods Ten healthy male Guangxi Bama miniature pigs,aged 4 to 5 months old,were selected and divided into control group and model group according to the random number table method,with 5 pigs in each group.After 12 hours of fasting in the two groups,the DCM model was replicated by intravenous injection of streptozotocin(STZ)150 mg/kg;for the Bama miniature pigs in the control group,citric acid-sodium citrate buffer 150 mg/kg was injected intravenously.After 10 months of modeling,the basic conditions of the two groups of animals were observed and their fasting blood glucose(FPG)levels were detected.The protein expression levels of NF-κB p65,TGF-β1,Bcl-2 and Bax in myocardial tissue of two groups were detected by Western Blot and the pathological changes of myocardial tissue were observed under electron microscope.Results In the model group,4 models were successfully established,and 1 died.The model pigs had symptoms such as polydipsia,polyphagia,polyuria and decreased body weight.The FPG level in the model group was significantly higher than that in the control group(mmol/L:25.53±3.75 vs.4.68±0.77,P<0.01).Compared with the control group,the protein expression levels of NF-κB p65,Bax and TGF-β1 in the myocardial tissue of model group were significantly increased(NF-κB p65/GAPDH:0.46±0.05 vs.0.38±0.02,Bax/GAPDH:0.46±0.01 vs.0.35±0.01,TGF-β1/GAPDH:0.39±0.01 vs.0.33±0.01,all P<0.05)and the expression level of Bcl-2 protein was significantly decreased(Bcl-2/GAPDH:0.33±0.01 vs.0.42±0.01,P<0.01).Electron microscopy results showed that the myofibrils of myocardial tissue in the DCM model group were disordered,and the number of mitochondria in the gap was significantly reduced.A large number of mitochondria with vacuolar degener

关 键 词：巴马小型猪 糖尿病心肌病模型 链脲佐菌素 核转录因子-κB p65 转化长长因子-β1 Bcl-2 Bax 

分 类 号：R54[医药卫生—心血管疾病]