6-姜酚靶向结合PRKACG调节PKA活性改善代谢相关脂肪性肝病的分子机制

代谢相关脂肪性肝病（MAFLD）是全球范围内普遍发病的一种慢性肝脏代谢性疾病，目前尚无针对性治疗MAFLD的化学合成药物。课题组研究发现中药生（干）姜中有效化学成分6-姜酚具有改善MAFLD的潜在药理价值，但其直接作用靶点及相关分子机理尚不清楚。鉴于此，本课题前期利用“标记”与“非标记”等靶标鉴别方法发现6-姜酚具有靶向结合PRKACG调节PKA活性的潜在分子机制。在此基础上，本课题拟采用蛋白质热漂移、竞争性结合及表面等离子共振等实验证实6-姜酚与PRKACG的直接结合；其次利用RNA干扰、质粒DNA转染等技术敲低或过表达PRKACG，考察其对6-姜酚调节PKA活性及介导的信息传递与脂质代谢生物学表型变化过程的影响，最终回归动物水平予以验证。总之，本课题的有效实施，对阐明6-姜酚改善MAFLD的直接作用靶点及“精细”分子机制具有重要意义，同时为中药药理作用机制研究提供新的思路与方法借鉴。

Metabolic associated fatty liver disease (MAFLD) is a widespread incidence chronic metabolic liver disease in the world, while there are no chemosynthetic drugs for the treatment of NAFLD presently. Our research group found that 6-gingerol, an effective chemical component in ginger, has a potential pharmacological value to treat MAFLD, but its direct target and related molecular mechanism remain unclear. And in the early stage of this project, we found that 6-gingerol could target binding to PRKACG to regulate PKA activity by using "labeled" and "non-labeled" target identification methods. On this basis, we intend to use protein thermal drift, competitive binding and surface plasmon resonance experiments to confirm the direct binding of 6-gingerol to PRKACG. Then, RNA interference, DNA plasmid transfection and other techniques are used to knockdown or overexpress PRKACG, and evaluate its effect on PKA activity, information transmission that 6-gingerol-mediated and the biological phenotype change of lipid metabolism, which will be verified by animal experiments. In conclusion, the effective implementation of this project is of great significance to clarify the direct target and "meticulous" molecular mechanism of 6-gingerol in improving MAFLD. Meanwhile, providing a new idea and method for the research on pharmacological mechanism of Traditional Chinese Medicine.