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目的 研究小鼠去泛素化酶卵巢肿瘤结构域6B(OTUD6B)的肝脏特异性敲降重组腺相关病毒（AAV）甲状腺素结合球蛋白（TBG）(AAV-TBG-shOTUD6B)在C57BL/6J小鼠肝脏中的敲降效率及其对肝脏糖代谢的调控作用。方法 设计并验证有效的siOTUD6B的序列，将非特异性干扰RNA阴性对照（shNC）和shOTUD6B的DNA序列插入含有启动子TBG的AAV载体骨架，构建并纯化得到AAV-TBG-shNC和AAV-TBG-shOTUD6B病毒。将AAV-TBG-shNC和AAV-TBG-shOTUD6B经尾静脉注射靶向小鼠肝脏，高脂饮食（HFD）喂养8周后检测小鼠OTUD6B蛋白和mRNA水平以及血糖水平、葡萄糖耐量、胰岛素敏感性、磷酸化蛋白激酶B(p-AKT)和磷酸化糖原合成酶激酶-3β(p-GSK-3β)蛋白表达水平。结果 与AAV-TBG-shNC组相比，AAVTBG-shOTUD6B组小鼠肝脏OTUD6B蛋白水平下降，小鼠肝脏OTUD6B mRNA水平下降（P<0.01）、小鼠禁食4 h血糖水平和禁食16 h血糖水平均降低（P<0.05,P<0.01）、小鼠肝脏p-GSK-3β蛋白水平降低（P<0.05），葡萄糖耐量增强（P<0.05）、胰岛素敏感性增强（P<0.01）、小鼠肝脏p-AKT水平升高（P<0.05）。结论 OTUD6B在影响肝脏糖代谢稳态中发挥重要作用，肝脏特异性敲降OTUD6B可改善HFD诱导的小鼠血糖水平，增强胰岛素敏感性。
Abstract：Objective To investigate the knockdown efficiency of the liver-specific knockdown recombinant adenoassociated virus(AAV) thyroid-binding globulin(TBG)(AAV-TBG-shOTUD6B) targeting the ovarian tumor do‑main 6B(OTUD6B) of the ubiquitin-proteasome system in the liver of C57 BL/6J mice and its regulatory effect on liver glucose metabolism.Methods An effective siOTUD6B sequence was designed and verified. The DNA se‑quences of non-specific interference RNA negative control(shNC) and shOTUD6B were inserted into the AAV vec‑tor backbone containing the promoter TBG to construct and purify AAV-TBG-shNC and AAV-TBG-shOTUD6B vi‑ruses. AAV-TBG-shNC and AAV-TBG-shOTUD6B were injected into mice via the tail vein to target the liver. Af‑ter 8 weeks of high-fat diet(HFD) feeding, the levels of OTUDB protein and mRNA in the mice liver, as well as blood glucose levels, glucose tolerance, insulin sensitivity, and protein expression levels of phosphorylated protein kinase B(p-AKT) and phosphorylated glycogen synthase kinase-3β(p-GSK-3β) were detected.Results Com‑pared with the AAV-TBG-shNC group, the OTUD6B protein level in the liver of the AAV-TBG-shOTUD6B group decreased, the mRNA level of OTUD6B in the liver decreased(P<0. 01), the fasting blood glucose levels of the mice after 4 hours of fasting and 16 hours of fasting both decreased(P<0. 05, P<0. 01), the protein level of pGSK-3β in the liver decreased(P<0. 05), glucose tolerance was enhanced(P<0. 05), insulin sensitivity was en‑hanced(P<0. 01), and the protein level of p-AKT in the liver increased(P<0. 05).Conclusion The construc‑tion of a recombinant adeno-associated virus for liver OTUD6B knockdown is successfully completed. OTUD6B plays an important role in regulating the homeostasis of liver glucose metabolism. Liver-specific knockdown of OTUD6B can improve the blood glucose levels of mice induced by HFD and enhance insulin sensitivity.
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基本信息:
DOI：10.19405/j.cnki.issn1000-1492.2026.09.001
中图分类号:R587.1;R-332
引用信息:
[1]时玉男,王凯,檀廷超,等.小鼠OTUD6B敲降腺相关病毒在肝脏糖代谢中的作用[J].安徽医科大学学报,2026(09):1505-1512.DOI:10.19405/j.cnki.issn1000-1492.2026.09.001.
基金信息:
国家自然科学基金项目(编号:82300659); 安徽省自然科学基金立项项目(编号:2308085QH247); 安徽医学高等专科学校2024年度省高校科研一般项目(编号:ZR2024B006); 安徽第二医学院科研创新团队项目(编号:WJH2023TLB001)，安徽第二医学院引进高层次人才科研启动基金项目(编号:2024RC011)
2026-08-07
2026-08-07
2026-08-07