基于NF-κB从油茶枯中发现有效抗炎、抗肿瘤活性成分的研究

Oil tea (Camellia oleifera Abel.) is one of the main economic forest species in south China, which has more than 2300 years of cultivation history. The fruit of this plant are used to produce Camellia oil. However, three times of Camellia cake was produced as byproduct, which is always discarded as waste residues or traded at low price. Although Camellia cake resource is abundant in our country, it has not been utilized comprehensively at present. In this project, all kinds of modern skills for separation and structural determination are employed to isolate and identify the chemical constituents of the effective fraction from Camellia cake monitored by LPS-induced NF-κB activation in RAW 264.7 macrophages. Based on this study, the isolated compounds will be screened for anti-inflammatory and anti-tumor activities in vitro. The aim of this study is to discover the anti-inflammatory and anti-tumor constituents from Camellia cake, especially those with both nice anti-inflammatory and anti-tumor activities. For the compounds with excellent pharmaceutical efficacy, molecular biological and pharmacological methods such as ELISA, RT-PCR, Western Blot will be applied to explore their possible anti-inflammatory and anti-tumor mechanisms. In a word, the task of this research is to explore the chemical constituents of Camellia cake through systematic separation and structural characterization, as well as their pharmarcological activites and the possible mechanisms. This research may provide theoretical and experimental data for further exploiting the abundant Camellia cake resource of our country.

油茶是我国南方的主要经济林品种之一，在我国已有2300多年的种植历史。油茶枯是茶油制取的副产物，其产量是茶油的3倍，资源丰富、贮量大，然而却常被废弃或低价买出，未能得以很好的综合利用，造成资源浪费。本项目以我国丰富廉价的油茶枯资源为研究对象，利用LPS诱导巨噬细胞RAW 264.7活化NF-κB建立初步的药理筛选模型，从油茶枯中获得具有NF-κB抑制活性的部位，再利用各种现代提取分离和结构测试手段从中获得有效单体化合物，在此基础上通过系统的抗炎、抗肿瘤活性筛选，发现油茶枯中有效的抗炎、抗肿瘤活性成分。对于药效显著，具有良好发展前景的化合物，运用ELISA、RT-PCR、Western blot等分子生物学和药理学的研究手段探讨化合物发挥抗炎、抗肿瘤活性的作用机制。通过油茶枯化学成分的分离鉴定、活性筛选和作用机制的研究，为我国油茶枯资源的深度研究与开发利用提供实验依据和理论指导。

本项目采用药理学和植物化学相结合的研究手段，明确了油茶枯乙醇提取物的乙酸乙酯、正丁醇和水溶性部位为其主要药效部位，采用各种现代分离手段从中分离鉴定了其中的33个小分子化合物的结构，并对2个新的多糖类成分进行了系统的研究，确定了油茶枯有效部位的化学成分。通过对油茶枯抗炎、抗肿瘤、免疫调节等药理活性的筛选，筛选出其中具有良好抗炎、抗肿瘤、免疫调节活性的成分，确立了油茶枯发挥药理活性的主要物质基础。对于油茶多酚、皂素、多糖这些在抗炎、抗肿瘤、免疫调节等药理活性上具有良好发展前景的代表性化合物或成分，运用了ELISA、Western blot、RT-PCR等分子生物学和药理学的研究手段，探索这些成分发挥药理活性的作用机制。摸索获得了一套从油茶枯中综合提取油茶多酚、山奈酚、皂素等成分的实用工艺，顺利完成了中试放大实验，具有高效、廉价、稳定、易于产业化放大等优点。通过油茶枯化学成分的分离鉴定、活性筛选、作用机制和活性成分提取工艺的研究，为油茶资源的深度研究与综合利用提供实验依据和理论指导。项目已基本完成了原申请书中预定的研究内容，并有所拓展；已在国家核心期刊以上杂志发表论文5篇；在国内学术会议论文集上发表会议论文3篇；国际学术会议上做邀请报告1次；申请国家发明专利2项；部分研究结果尚待整理于后期发表。在人才培养上，项目培养硕士研究生3名，2名已获硕士学位，1名在读；项目负责人和项目组成员经过此项目的开展在科研能力等方面得到了较大程度的提高。