基于脂联素的多聚化修饰探讨祛湿活血方治疗非酒精性脂肪肝的分子机制

脂联素是目前发现的唯一有益于非酒精性脂肪肝（NAFLD）的细胞因子，高分子量（HMW）脂联素是其主要活性形式，其多聚化修饰主要受分子伴侣DsbA-L调节。我们的工作和已有的研究发现，祛湿活血方能促进脂肪细胞HMW脂联素以及脂肪细胞DsbA-L的表达；脂肪细胞内DsbA-L的表达受AMPK和Akt/FOXO1通路调节，脂肪细胞外，受TNF-α调节。本项目拟从以上调控DsbA-L途径的角度，探讨祛湿活血方在细胞内外对脂联素多聚化的调节机制。项目通过免疫组化、Westernblot、RNA干扰、腺病毒感染等手段，在NAFLD大鼠中观察中药处理后脂肪组织以上通路各分子的变化；以及在原代脂肪细胞和3T3-L1脂肪细胞系，阻断或活化以上各信号通路，观察祛湿活血方对DsbA-L/HMW脂联素表达的影响，以期阐明祛湿活血方调节脂联素多聚化表达的分子机制，可能对中医药靶向治疗NAFLD产生重要影响。

Adiponectin is the only beneficial cytokine for non-alcoholic fatty liver disease (NAFLD).High molecular weight (HMW) adiponectin is major activity form in vivo. Multimerization modification of adiponectin is regulated by molecular chaperones DsbA-L. Our early study has found Queshihuoxue (QSHX) decoction promoted significantly HMW adiponectin expression of serum and adipocyte, and promoted DsbA-L expression of adipocyte. Other studies found that DsbA-L expression is regulated by AMPK and Akt/FOXO1 pathways intra-adipocyte, and extra-adipocyte by TNF-α. This project will study molecular mechanism of regulating high molecular weight adiponectin by QSHX decoction basing on DsbA-L expression of intra- and extra-adipocyte. By way of immunohistochemistry stain, Westernblot, RNAi and adenovirus infection, the change of each upper-stream molecular of DsbA-L after QSHX decoction treatment in NAFLD rats fatty tissue, and the effect of QSHX decoction on DsbA-L/adiponectin expression after blocked or activated above pathway in primary adipocytes and 3T3-L1 line, will be observated. This project is to illuminate the molecular mechanism of QSHX decoction regulating HMW adiponectin and to establish study basis of mechanism for traditional decoction treating NAFLD.