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用贵州小香猪建立 2型糖尿病动物模型 ,探讨糖尿病小型猪三磷酸腺苷结合盒转运体A1表达的变化。采用高脂高蔗糖饲料喂养贵州小香猪 ,建立 2型糖尿病动物模型。血浆总胆固醇、甘油三酯、高密度脂蛋白胆固醇和葡萄糖的浓度均用氧化酶法测定 ,血浆游离脂肪酸用比色法测定 ,采用逆转录—聚合酶链反应、Western印迹和免疫组织化学法分别检测三磷酸腺苷结合盒转运体A1mRNA和蛋白质的表达。喂养 6个月后 ,实验组与正常对照组比较 ,空腹血糖值明显升高 ;空腹胰岛素水平在头 3个月轻度升高 ,在第 6个月末其水平降低 ;血清总胆固醇、甘油三酯和游离脂肪酸水平升高。采用逆转录—聚合酶链反应检测组织细胞三磷酸腺苷结合盒转运体A1mRNA的表达 ,结果显示 ,与对照组小型猪比较 ,实验组小型猪肝组织、冠状动脉和肾组织三磷酸腺苷结合盒转运体A1mRNA表达上调 (P <0 .0 5 )。Westernblot检则肝组织、冠状动脉和肾组织三磷酸腺苷结合盒转运体A1蛋白质的表达 ,结果显示 ,与对照组小型猪比较 ,高脂高蔗糖饲料喂养的小型猪肝组织、冠状动脉和肾组织三磷酸腺苷结合盒转运体A1蛋白质表达明显增高 (P <0 .0 5 )。免疫组织化学检测的结果与Westernblot检则结果相一致 ,实验组小型猪肝组织、冠状动脉和肾组织切片上棕褐色或棕黄色
Animal models of type 2 diabetes in Guizhou miniature pigs were established to investigate the changes of adenosine triphosphate binding cassette transporter A1 in diabetic miniature pigs. Guizhou high-fat pigs were fed with high-fat and high-sucrose diet to establish animal model of type 2 diabetes. Plasma total cholesterol, triglycerides, high-density lipoprotein cholesterol and glucose concentrations were measured by oxidase method, plasma free fatty acids by colorimetric assay, reverse transcription - polymerase chain reaction, Western blot and immunohistochemistry Detection of adenosine triphosphate binding cassette transporter A1 mRNA and protein expression. Compared with the normal control group, the fasting blood glucose level was significantly increased in the experimental group at 6 months after feeding. The fasting insulin level increased slightly in the first 3 months and decreased at the 6th month. The serum total cholesterol, triglyceride And free fatty acid levels increased. The expression of adenosine triphosphate binuclear transporter A1 mRNA was detected by reverse transcriptase-polymerase chain reaction. The results showed that compared with the control group, the A1 mRNA expression of adenosine triphosphate binuclear transporter in liver tissue, coronary artery and kidney of experimental group was up-regulated (P <0. 05). Western blot analysis of liver tissue, coronary artery and kidney adenosine triphosphate box transporter Al protein expression, the results showed that compared with the control group of miniature pigs, high fat and high sucrose diet of small pig liver tissue, coronary artery and kidney adenosine triphosphate binding Box transporter A1 protein expression was significantly increased (P <0.05). The result of immunohistochemistry was consistent with the result of Western blot test. The experimental group of small pig liver tissue, coronary artery and kidney tissue sections were tan or brown