glp1通过wnt通路改善肝脏胰岛素抵抗机制探讨-study on the mechanism of gl p1 improving hepatic insulin resistance through wnt pathway.docx

独创性声明本人郑重声明，本学位论文是本人在导师指导下进行的研究工作及取得的研究成 果的总结。尽我所知，除文中已经标明引用的内容外，本论文不包含任何其他个人或 集体已经发表或撰写过的研究成果。对本文的研究做出贡献的个人和集体，均已在文 中以明确方式标明。本人完全意识到本人将承担本声明引起的一切法律后果。学位论文作者签名： 日期：年月日学位论文版权使用授权书本学位论文作者完全了解学校有关保留、使用学位论文的规定，即：学校有权保 留并向国家有关部门或机构送交论文的复印件和电子版，允许论文被查阅和借阅。本 人授权华中科技大学可以将本学位论文的全部或部分内容编入有关数据库进行检索， 可以采用影印、缩印或扫描等复制手段保存和汇编本学位论文。保密□ ，在年解密后适用本授权书。 本论文属于不保密□。（请在以上方框内打“√” ）学位论文作者签名：指导教师签名：日期：年月日日期：年月日目 录中文摘要 ··········································································· 1英文摘要 ··········································································· 2研究背景和目的 ·································································· 4材料与方法 ········································································ 6结果 ················································································19讨论 ················································································30 结论 ················································································33 参考文献 ··········································································34 综述 ················································································38 参考文献 ··········································································43 致谢 ················································································47附录 ················································································48GLP-1 通过 WNT 通路改善肝脏胰岛素抵抗机制探讨华中科技大学同济医院内分泌科 硕士研究 生：秦瑜 指导教师：袁刚教授 中文摘要目的：研究 GLP-1 对糖尿病小鼠的作用及其改善肝脏胰岛素抵抗可能的分子生物学机制。方法：将 db/db 小鼠分为两组，一组为生理盐水组（对照），一组为利拉鲁肽组，同 时在相同的饲养环境下饲养。两组动物每天分别腹腔注射生理盐水与利拉鲁肽，干预 8 周，检测小鼠的体重，血糖，胰岛素和 HOMA-IR。取肝脏石蜡包埋固定，H-E 染色 观察。提取动物肝脏的蛋白和 RNA，用 Western Blots 和 real-time PCR 方法检测糖异 生酶与 WNT 通路相关蛋白与 RNA 的表达情况。与此同时在体外用 FFA 干预 HepG2 细胞，建立胰岛素抵抗模型。利用 siRNA 转染技术使β-catenin 沉默，不同利拉鲁肽 浓度 10-8mol/L，10-7mol/L 干预细胞，Western Blots 检测 p-FoxO1，PEPCK，G6Pase 和 WNT 信号通路相关蛋白的表达水平。GOD-POD 法检测培养基中葡萄糖剩余量。 结果：与对照组相比，利拉鲁肽组干预四周开始，体重显著减轻，血糖下降，胰岛素 水平减低，HOMA-IR 值降低，且作用一直持续到实验结束。利拉鲁肽治疗后肝脏结 构改善，脂滴减少。利拉鲁肽组肝脏内的 G6Pase 与 PEPCK 的表