T2DM小型猪模型骨骼肌、肝脏及胰腺组织蛋白激酶B表达及磷酸化的变化

Changes of protein kinase B and phosphorylation expression in the skeletal muscle,liver and pancreas of miniature pig model with type 2 diabetes mellitus model

目的研究高脂高糖饮食联合链脲佐菌素(STZ)构建巴马小型猪2型糖尿病(T_2DM)模型的可行性,并观察小型猪模型骨骼肌、肝脏及胰腺组织蛋白激酶B(PKB)表达及磷酸化变化。方法将健康雄性巴马小型猪10只随机分为2组:对照组5只,喂养普通饲料;糖尿病模型组5只,高脂高糖饮食联合STZ构建巴马小型猪T_2DM模型。检测2组小型猪空腹血糖、空腹胰岛素水平,并计算胰岛素抵抗指数(HOMA-IR)。采用Western blot法检测骨骼肌、肝脏及胰腺组织中PKB蛋白表达及磷酸化水平。结果 (1)高脂高糖饲料喂养10个月后,与对照组比较,模型组体质量、空腹血糖、空腹胰岛素水平及HOMA-IR明显升高(P<0.01)。(2)与对照组相比,模型组STZ给药后空腹血糖水平进一步升高,空腹胰岛素水平明显下降(P<0.01)。(3)与对照组比较,模型组骨骼肌、肝脏及胰腺组织中PKB蛋白表达及磷酸化水平明显降低(P<0.05)。结论高脂高糖饮食联合STZ多次腹腔注射可以成功构建T_2DM小型猪模型,模型组小型猪骨骼肌、肝脏及胰腺组织PKB蛋白表达及磷酸化水平明显降低。

Objective To study the feasibility of Bama miniature pig model establishment of type 2 diabetes mellitus （T2DM） by high-fat and high-sugar diet combined with streptozotocin （STZ）, and observe the changes in protein kinase B （PKB） and phosphorylation expressions in skeletal muscle, liver, and pancreatic tissues of the miniature pig model. Methods A total of 10 healthy male Bama miniature pigs were randomly divided into two groups：control group （n=5, fed normal diet） and diabetic model group （n=5, fed high-fat and high-sugar diet combined with STZ to establish T2DM Bama miniature pig model）. The fasting plasma glucose （FPG） and fasting insulin （FINS） levels were measured, and insulin resistance （HOMA-IR） was calculated in two groups. The PKB and phosphorylation expressions in skeletal muscle, liver and pancreatic tissues were measured using Western blot assay. Results （1） After 10 months of high-fat and high-sugar diet, the body weight, FPG, FINS and HOMA-IR were significantly higher in model group than those of control group （P〈0.01）. （2） After STZ treatment, compared with control group, there was a further increased level of FPG and a significantly decreased level of FINS in model group （P〈0.01）. （3） Compared with control group, the PKB and phosphorylation expression levels in skeletal muscle, liver and pancreas were significantly lower in model group （P〈0.05）. Conclusion The high-fat and high-sugar diet combined with STZ can successfully establish the T2DM miniature pig model. The PKB and phosphorylation expression levels in skeletal muscle, liver, and pancreatic tissues are decreased in model pigs.