黄花败酱改善3T3-L1脂肪细胞胰岛素抵抗的活性成分及作用机制研究

Diabetes mellitus seriously affects people's life and health because of its high morbidity and mortality. Its pathogenesis is complexed and now it is generally believed to be insulin resistance, which may relate to inflammatory response. Thus, it has been a popular research direction to look for antidiabetic drugs from natural products under the guidance of improving inflammation and insulin resistance. Previous studies have found that five compounds from the EtOAc extract of Patrinia scabiosaefolia could significantly promote glucose uptake. Among theme, patrinoside (10) and the new compoud 13 could activate PI3K and Akt signaling to mediate glucose uptake. However, their effects on inflammation, other compounds with better activities in n-butanol extract, structure-activity relationship and their mechanisms are worthy to make further studies. Hence, in this project, the substantial basis from n-butanol extract will be identified systematically to find compounds with better activity. Their structure-activity relationship and mechanisms will be discussed under the guidance of improving inflammation and insulin resistance in 3T3-L1 adipocytes. The implementation of this project may obtain anti-diabetic lead compounds, and illuminate the active ingredients on improving insulin resistance of P. scabiosaefolia and their mechanisms, and then provide scientific basis for the development and effective use of P. scabiosaefolia.

糖尿病因其高发病率和高死亡率严重影响人们的生活健康，其发病机制复杂，目前普遍认为是胰岛素抵抗，而炎症可能是引起胰岛素抵抗的重要因素。以改善炎症与胰岛素抵抗为方向，从天然产物中寻找抗糖尿病药物，成为研究的热点。前期研究发现黄花败酱提取物具有抗胰岛素抵抗活性；目前从乙酸乙酯部位中发现5个化合物可显著促进葡萄糖摄取，其中patrinoside (10) 和新化合物13可激活PI3K/Akt信号通路调控葡萄糖摄取。但活性更好正丁醇部位是否存在活性更好的化合物及构效关系、作用机制等值得深入研究。因此，本项目拟在前期工作基础上，继续开展活性部位物质基础研究，以影响胰岛素抵抗葡萄糖摄取和炎症反应为方向，寻找活性更强的分子，探讨初步构效关系，并阐明强活性分子的作用机制。本项目的实施有望获得抗糖尿病药物的先导化合物，同时阐明黄花败酱改善胰岛素抵抗的活性分子及作用机制，进而为开发利用黄花败酱植物提供科学依据。

为了寻找黄花败酱中改善胰岛素抵抗活性成分及作用机制，该项目系统研究了黄花败酱改善胰岛素抵抗物质基础及强活性分子的作用机制。从黄花败酱植物中共分离鉴定化合物91个，以环烯醚萜类化合物和木脂素类化合物为主，其中新化合物18个。对分离得到的化合物，在3T3-L1脂肪细胞上进行改善胰岛素抵抗葡萄糖摄取活性和在RAW264.7细胞上进行抗炎活性评价，共发现17个具有不同程度的降糖活性化合物，其中4个化合物还具有显著抗炎活性。采用Western blot方法进一步研究活性化合物作用机制。结果发现，化合物5、6、43、57和59可通过激活PI3K/AKT信号通路改善胰岛素抵抗。同时化合物5和化合物6还可以抑制NF-κB和MAPK通路的激活，抑制炎症因子和趋化因子的分泌及抑制巨噬细胞向脂肪细胞浸润，共同发挥改善胰岛素抵抗作用。该项目的完成，不仅阐明了黄花败酱的化学成分，丰富了环烯醚萜类化合物的多样性，而且明确了黄花败酱中改善胰岛素抵抗的主要活性分子及作用机制，为该属植物的进一步开发与利用提供坚实的物质基础和科学依据。