基于Notch信号通路探讨活血利水法对糖尿病肾病巨噬细胞活化介导肾小管上皮细胞转分化的影响

Promoting Blood Circulation and Alleviating Water Therapy is an important clinical syndrome differentiation and treatment to Diabetic Nephropathy (DN) of Traditional Chinese Medicine, the curative effect is positive. Inflammatory reaction mediated by Macrophage activation is important factor to DN development. Notch signal pathway plays a key function in the process. In our previous research we had demonstrated that Danggui Shaoyao Powder (DSS) was able to inhibit expression of Macrophage related inflammatory factors and renal tubular Epithelial-Mesenchymal Transition ,which is representative prescription of Promoting Blood Circulation and Alleviating Water Therapy. We put forward the hypothesis: The mechanism of Promoting Blood Circulation and Alleviating Water Therapy treatment of DN is to inhibit Macrophage activation by Notch signal pathway, followed by regulating Th1/Th2 balance and inhibiting renal tubular Epithelial-Mesenchymal Transition. In this study, the effect of DSS on the change of Macrophage activation, proliferation, Th1/Th2 balance, renal tubular Epithelial-Mesenchymal Transition and key molecular in Notch signal pathway at Diabetic kidney is studied by using Light Microscope, Real-time PCR, Western-Blot, ELISA, Cell Culture technology and Immunohistochemistry In Vitro and In Vivo.Explore the function of Promoting Blood Circulation and Alleviating Water Therapy reduce renal tubular Epithelial-Mesenchymal Transition mediated by macrophage activation through Notch signal pathway, Revealing the treatment mechanisms of Promoting Blood Circulation and Alleviating Water Therapy to DN, The study will provide new idea and method to Chinese medicine treatment of DN.

活血利水法是糖尿病肾病（DN）中医临床辨治重要治法，疗效确切，由巨噬细胞活化所介导的炎症反应是导致DN持续发展的重要因素，Notch信号通路在该过程中起关键作用。前期研究发现活血利水法代表方当归芍药散（DSS）有抑制DN巨噬细胞相关炎性因子表达和肾小管上皮细胞转分化作用。据此提出假说：活血利水法治疗DN的机制是通过Notch信号通路抑制巨噬细胞活化继而调节Th1/Th2失衡及抑制肾小管上皮细胞转分化。本研究通过体内外试验，应用光镜、Real-time PCR、WB、ELISA、免疫组化双染和细胞培养技术，研究DSS对DN巨噬细胞活化、Th1/Th2平衡、肾小管细胞转分化及Notch信号通路关键分子变化的影响，探讨活血利水法对Notch通路及巨噬细胞活化介导肾小管上皮细胞转分化作用，揭示活血利水法治疗DN的机制和作用环节，为中医药治疗糖尿病肾病提供新的思路和方法。

巨噬细胞活化介导的炎症反应及肾小管上皮细胞转分化与糖尿病肾病（DN）持续发展密切相关，Notch信号通路在该过程中起关键作用。活血利水法是DN中医临床辨治重要治法。本项目依据中医学“血不利则为水”理论，借助信号转导通路研究策略，以巨噬细胞活化与DN的共同调控点Notch信号通路为切入点，采用体内外实验，以早期糖尿病肾损伤动物模型和体外活化的巨噬细胞及肾小管上皮细胞为研究对象，研究活血利水法代表方当归芍药散对糖尿病大鼠肾组织巨噬细胞活化、肾小管上皮细胞转分化及Notch信号通路关键分子变化的影响。通过本项研究，证明活血利水法可能通过抑制巨噬细胞Notch信号通路，改善肾小管上皮细胞转分化延缓糖尿病进程。从而明确活血利水法的治疗机制，探讨血不利则为水理论在治疗早期糖尿病肾病的科学内涵，为开发治疗早期糖尿病肾病的新药提供依据。