糖尿病状态下CTRP13下调介导的肝窦毛细血管化的机制研究

糖尿病合并脂肪肝的形成与高糖诱导的肝窦毛细血管化有关。CTRP13是一个新的调节糖脂代谢的因子，参与了肝脏的脂肪变性。我们前期研究发现高糖可诱导肝窦内皮细胞CTRP13下调，然而，它在肝窦毛细血管化中的作用机制不清，国内外未见报道。本研究利用db/db小鼠模型和体外细胞实验，运用基因重组、siRNA、电镜、RT-PCR、WB、免疫组化、脂肪超声定量等技术，通过CTRP13过表达从动物水平研究CTRP13与肝脏肝窦血流、窗孔调节、基底膜形成及炎症因子变化的关系；在细胞水平研究高糖对肝窦内皮细胞CTRP13、PLVAP、AMPK、Rho 表达及摄糖摄脂功能的影响以及高糖/CTRP13介导的肝窦毛细血管化是否与CaMKK、AMPK、Rho途径有关；临床研究糖尿病患者CTRP13与糖脂代谢及脂肪肝的相关性。旨在阐明CTRP13介导的肝窦毛细血管化的分子机制，为糖尿病合并脂肪肝的防治提供理论依据。

The nonalcoholic fatty liver disease(NAFLD) was closely associated with hyperglycemia-induced hepatic sinusoid capillarization in diabetes. CTRP13 is a new regulated glucolipid metabolic factors, involved in hepatic steatosis. Our previous experiment found that high glucose induceddown-regulation of CTRP13 in hepatic sinusoidal endothelial cells. However, it remained unclear whether CTRP13 was involved in hepatic sinusoid capillarization. In this study, db/db mice model and cell experiment in vitro were established, genetic recombination, siRNA, electron microscope, RT-PCR, WB, immunohistochemical and fat quantitative techniques with ultrasound were used,respectively.In vivo, the relationship between CTRP13 expression and hepatic sinus blood flow, the characteristics of fenestrae, basement membrane formation and inflammatory factor were investigated. In vitro, the influence of high glucose on CTRP13, PLVAP, AMPK, Rho expression and glucolipid-taken function were determined, and whether high glucose/CTRP13 mediated hepatic sinusoid capillarization is related to CaMKK, AMPK and Rho. In clinical studies, the correlation of CTRP13 and glucolipid metabolism and fatty liver were explored in type 2 diabetes mellitus(T2DM). This study aimed to clarify the molecular mechanisms of CTRP13 mediated hepatic sinusoid capillarization in daibetes, and to provide theoretical basis for the prevention and treatment of NAFLD in T2DM.