【作者】 赵静；

【导师】 葛鹏玲；

【作者基本信息】 黑龙江中医药大学 ， 药理学， 2015， 硕士

【摘要】 目的:探讨db/db小鼠在瘦素受体缺失条件下引起2型糖尿病胰岛素抵抗的分子机制,为开发防治2型糖尿病胰岛素抵抗的药物提供可能的靶点。方法:①不同周龄雄性db/db小鼠(3周龄、6周龄、9周龄),测定相应模型组及其正常对照组的血糖值、胰岛素值、体重、胰岛素敏感指数等相应2型糖尿病胰岛素抵抗指标。②利用全基因组表达谱芯片技术初步筛选3周龄db/db小鼠肝组织中差异表达的基因;③应用Gene Ontology数据库整合分析差异表达基因的功能及其可能参与调控的生物学过程。④对选取的差异显著的基因进行实时荧光定量PCR(qRT-PCR)实验,验证基因芯片的数据结果。结果:①3周龄小鼠模型组与其正常对照组比较,血糖值和体重均正常,出现高胰岛素、低胰岛素敏感指数等特征;6周龄、9周龄小鼠模型组与相应正常对照组比较均出现肥胖、高血糖、高胰岛素、低胰岛素敏感指数的2型糖尿病现象。②基因芯片结果显示,差异表达显著的基因共197条,其中表达上调的基因93条,表达下调的基因有104条;③对差异表达基因进行GO功能分析,发现基因EGFR、CNTF参与GO富集度得分较高的磷酸化调节过程,ATP5O、GLRX2参与GO富集度得分较高前体代谢及能量产生过程,差异表达基因INSR、DYRK1A分别参与了与2型糖尿病可能相关的激活MAPK活性过程、蛋白磷酸化过程。④应用qRT-PCR实验验证上述6个基因芯片结果。结论:实验结果表明,3周龄db/db模型鼠已经存在早期2型糖尿病胰岛素抵抗症状。瘦素信号转导障碍条件下差异表达的基因EGFR、CNTF,ATP5O、GLRX2、INSR、DYRK1A与2型糖尿病胰岛素抵抗发病机制密切相关。更多还原

【Abstract】 Objective:Explore the molecular mechanism of the insulin resistance to type 2 diabetes under the condition on the lack of leptin receptor of the db/db mice,and development the possible targets for type 2 diabetes drugs of early prevention and treatment.Methods:①Different weeks of male db/db mice(3 weeks,6 weeks,9 weeks),determination of the corresponding model group and normal control group,weight,blood-glucose,insulin value and insulin sensitive index of the type 2 diabetes index.②Using genome-wide expression profile chip technology Determined the differentially expressed gene on 3 weeks of the db/db mice liver tissue.③Application of Gene Ontology database,and analysis of differentially expressed genes involved in the regulation of biological processes.④Detectioning the differentially expressed genes on the real-time quantitative PCR(qRT-PCR)experiment,and verify the result of gene chip data.Results:①3 weeks of mice model group and normal control group,blood glucose and weight are normal,but have the high insulin value,and low insulin sensitive index characteristic;6 weeks and 9 weeks of age mice model group compared with the corresponding normal control group on obesity,high blood glucose,high insulin value,low insulin sensitive index of type 2 diabetes;②Gene Microarray results,the differentially expressed genes were 197 genes,of which 93 genes were up-regulated and 104 genes were down-regulated;③GO function analysis of differentially expressed genes,found that the gene EGFR and CNTF Participate in the regulation of phosphorylation process;ATP50 and GLRX2 Participate in the generation of precursor metabolites and energy process,gene INSR and DYRK1A were separately related to the activation of MAPK activity and the protein phosphorylation which may be involved in type 2 diabetes④Application qRT-PCR to verify the the six differentially expressed genes.Conclusion:Experimental results show that three weeks of db/db model rat has existed phenomenon of early insulin resistance.Under the leptin signal transduction disorder,the significantly differentially expressed genes of EGFR,CNTF,ATP50,GLRX2,INSR,DYRK1A may closely associated with the pathogenesis of insulin resistance to type 2 diabetes.更多还原