基于miRNA调控SIRT1/NF-κB信号通路探讨滋阴清热法治疗2型糖尿病胰岛素抵抗的分子机制

Insulin resistance(IR) is the driving factor in the occurrence and progression of Type 2 Diabetes Mellitus，running through the whole journey. Our research group have many years’ experience on taking the method of nourishing yin and clearing heat to treat IR of type 2 diabetes，and the effect was significant by treating diabetic insulin resistance with nourishing yin and clearing heat method. Prophase research shows that Qing run formula mainly treats diabetic insulin resistance by improving inflammation state of the liver. The new research suggests that the pathway of miR-34a/ miR-181a -SIRT1/NF- kappa B in the liver inflammation is closely related to IR of type 2 diabetes. we speculate that Qing run formula may improve inflammation state to treat diabetic insulin resistance by negativing regulatory the expression of miR-34a/ miR-181a thus activating the signal pathway of SIRT1/NF- kappa B.With the pathway of miR-34a/ miR-181a -SIRT1/NF- kappa B flowing as the starting point through Diabetic Rats and IR model of HepG2 cell ,this study plans to investigate molecular mechanism of Qing run formula treating diabetic insulin resistance while it has important significance in the inheritance and excavation of the experiences of some famous veteran doctors of TCM and further reveal the scientific connotation of the theory thus providing new ideas and targets to treating diabetic insulin resistance in clinic.

胰岛素抵抗（IR）是2型糖尿病发生和进展的驱动因素，并贯穿于其全程。本课题组多年来采用清润方代表的滋阴清热法治疗2型糖尿病IR，疗效确切。前期研究表明，清润方主要通过改善肝脏的炎症反应来治疗IR。最新研究提示miR-34a/ miR-181a介导的肝内SIRT1/NF-κB炎症通路和2型糖尿病IR密切相关。我们推测清润方可能是通过抑制miR-34a/ miR-181a的表达，从而激活SIRT1/NF-κB信号通路，改善炎症状态来治疗糖尿病IR。本研究拟通过2型糖尿病大鼠及HepG2细胞的IR模型，以miR-34a/ miR-181a-SIRT1/NF-κB通路为切入点，应用qRT-PCR、Western blot、siRNA等技术，体内外实验相结合，探讨清润方治疗2型糖尿病IR的机制，对传承、挖掘名老中医经验，揭示其理论科学内涵具有重要意义，为治疗2型糖尿病IR提供新思路和靶标。

2型糖尿病（Type 2 Diabetes Mellitus，T2DM）严重危害人类健康，是我国慢性病的防控重点。胰岛素抵抗（Insulin，resistance，IR）是T2DM发生和进展的驱动因素，积极改善IR成为治疗T2DM的关键策略。本课题组应用滋阴清热法治疗T2DM-IR已有多年的工作积累。本研究开展了在体动物实验和离体细胞实验两部分研究，利用Western blot、qRT-PCR、ELISA、siRNA等技术探讨了清润方代表的滋阴清热法改善T2DM-IR的作用及机制。体内实验以高脂饮食联合链脲佐菌素诱导SD大鼠建立T2DM大鼠模型，发现清润方可减轻T2DM大鼠糖脂代谢紊乱，改善IR状态，缓解肝脏病理损伤，并下调肝组织miR-181a、miR-34a、NF-κB的表达水平，上调SIRT1、GLUT4的表达水平。体外实验以高浓度胰岛素诱导HepG2细胞建立IR细胞模型，以清润方及其有效组分干预后，发现清润方及其有效组分可提高IR-HepG2细胞的葡萄糖摄取能力，改善IR状态。清润方代表的滋阴清热法可通过抑制miR-181a、miR-34a的表达，激活SIRT1/NF-κB信号通路，下调TNF-α、IL-6的表达，改善炎症状态从而改善T2DM-IR。