基于ADK/Adenosine调控DNA甲基化探讨“利湿化瘀通络”法对2型糖尿病肾病足细胞裂孔膜损伤的干预机制研究

腺苷激酶途径（ADK/Adenosine）调控DNA甲基化介导足细胞裂孔膜(SD)损伤是2型糖尿病肾病(T2DKD)的病理新机制，目前尚无理想治疗方法。基于中医对T2DKD “湿瘀阻络”基本病机的认识，我们证实“利湿化瘀通络”消肾方对T2DKD蛋白尿和肾脏损伤有显著临床疗效。在此基础上，本课题拟利用eNOS(-)/(-)db/db小鼠、C57BLKS/Jdb/db小鼠两种T2DKD模型和离体培养足细胞，应用MeDIP-chip和MSP等DNA甲基化检测手段，免疫荧光双标、ChIP、基因过表达或knockdown等分子生物学技术和中药血清药理学方法，以ADK/Adenosine调控DNA甲基化为切入点，探讨消肾方对T2DKD足细胞SD损伤的保护机制。本研究有助于阐明消肾方治疗作用的物质基础和科学内涵，明确中医“利湿化瘀通络”法通过干预表观遗传学机制治疗T2DKD蛋白尿和肾脏损伤的特色和优势。

The adenosine kinase pathway (ADK/Adenosine) regulating podocytes DNA methylation plays a pivotal role in mechanisms for slit diaphragm (SD) damages and proteinuria of Type 2 Diabetic kidney disease (T2DKD). There is no effective method to intervene this mechanism till now. Based on the pathogenesis “Collaterals blockage by dampness and stagnancy” of Chinese Medicine about DKD, we have confirmed that Xiao-Shen-Formula (XSF) with the effect of “Removing dampness, dissolving stagnancy and relaxing collaterals” had remarkable clinical therapeutic effect on proteinuria and renal injury in T2DKD patients. On this basis, eNOS(-)/(-) db/db , C57BLKS/J db/db mice, the cultured podocytes, the multiple molecular biological methods such as MeDIP-chip, MSP, knock down, over expression, chromatin immunoprecitation (ChIP) and Chinese medicine serum pharmacology method will be adopted to investigate the therapeutic effect of XSF on ADK/Adenosine regulating podocytes DNA methylation and podocytes SD damages in T2DKD. The present study aim to explore the experimental basis and scientific evidence for clinical usage of XSF. From the point of regulating epigenetics mechanisms, the advantage and unique feature of Chinese Medicine “Removing dampness, dissolving stagnancy and relaxing collaterals” in treating proteinuria and renal injury of T2DKD will be further investigated.