万年蒿抑制脂肪蓄积活性成分作用机理及其协同作用研究

Artemisia sacrorum Ledeb. (Compositae), which is one kind of Korean Minority folk medicine that has been traditionally used to prevent and treat fatty liver and other hepatic diseases, has a long and wide application history. Based on the confirmation of the active part of attenuation on lipid accumulation in our previous work, this project intends to separate compounds from the active part by applying chromatographic methods such as HPLC, perparative HPLC, etc. and identify their chemical structures. And then in vitro experiments will be applied to find 3 compounds with best activities. After that, this project will adopt western blot analysis,RT-PCR,pathological observation and other methods,to comprehensively study the effects of 3 active compounds and their optimal prescription screened by orthogonal test on several lipid accumulation-related signal pathways such as 'AMPK- - ACC- - Malonyl CoA- - CPT1', etc. in cell and animal levels,by means of evaluation of the proteins and genes expressions of AMPK, SREBP1c, etc. and other techniques, so as to investigate their mechanisms and synergic effects of attenuative effects on lipid accumulation. If it is approved and then completed, this project will provide theoretical basis and technical support on treating fatty liver(endemic disease)and other lipid accumulation-related diseases.

万年蒿是一种中国朝药，常用其预防和治疗脂肪肝等各种肝脏疾病，有悠久、广泛的应用基础。在前期工作基本确定万年蒿抑制脂肪蓄积有效部位ASL的前提下，本项目拟应用HPLC等色谱学方法，从ASL中分离得到化学成分并进行结构鉴定，之后通过细胞实验对各成分的抑制脂肪蓄积活性进行测定，筛选出活性最好的三个化学成分。然后，采用Western blot analysis，RT-PCR等方法，通过评估细胞和肝组织中AMPK等蛋白质或基因表达等多种技术手段，在细胞和动物水平上对三个活性成分及通过正交试验筛选出来的最优组方进行全方位的研究，探讨其对"AMPK- - ACC- - Malonyl CoA- - CPT1"等多个与脂肪蓄积相关的信号途径的影响，以深入探究万年蒿对机体的整体协同作用，阐明其抑制脂肪蓄积作用机理。本项目的立项和完成将对应用万年蒿治疗地方性疾病脂肪肝及与脂肪蓄积密切相关的疾病提供理论基础和技术支持。

万年蒿是一种中国朝药，常用其预防和治疗脂肪肝等各种肝脏疾病，有悠久、广泛的应用基础。本项目在进一步确证了万年蒿抑制脂肪蓄积有效部位的前提下，应用HPLC等色谱学方法，从万年蒿有效部位中分离出十八个单体化合物并对其中完成了化学结构鉴定。综合各方面因素后，从万年蒿有效部位中筛选出十二个单体化合物，应用3T3-L1前脂肪细胞和HepG2细胞等实验继续进行进一步的筛选。实验结果表明，其中化合物W在多种实验中均显示了最强的抑制脂肪蓄积作用。因此，采用Western blot analysis，RT-PCR等方法，因此对其进行了深入的药效学确证以及作用机理研究，阐明了其调节AMPK，ACC，SREBP1c，FAS等蛋白质及基因的作用机理。另外，以化合物W为核心研究了万年蒿抑制脂肪蓄积最优组方，证明了协同作用的存在，并阐明了其协同作用机理。本项目的完成将对应用万年蒿治疗脂肪肝及与脂肪蓄积密切相关的疾病提供坚实的理论基础和技术支持。