新疆阿魏Ferula sinkiangensis K. M. Shen 属于伞形科阿魏属植物，主产于我国新疆伊犁等地，具有消积，化瘕，散痞，杀虫等作用，是我国传统的民族药。国内外大量研究表明，新疆阿魏具有显著的抗炎活性，被广泛应用于胃肠道疾病、类风湿性关节炎、支气管炎等炎症相关疾病的治疗。但由于自然环境的改变以及长期以来的乱采滥挖等因素，使其生长环境遭到极大破坏，导致阿魏资源越来越匮乏，新疆阿魏更是被列为我国二级濒危植物。目前只有新疆阿魏和阜康阿魏Ferula fukanensis K. M. Shen被收录在2020版《中国药典》中，其药用部位为树脂且只能通过切割根或茎产生，极难获得，而易获取的地上部位则常常被丢弃，这造成了极大的资源浪费。因此，若能将新疆阿魏地上部位变废为宝，从中发现新的天然活性成分，将对新疆阿魏的环境保护和资源利用起到极为重要的作用。本课题组前期通过体外抗炎活性筛选发现新疆阿魏地上部位95%乙醇提取物的二氯甲烷极性段对脂多糖(LPS)诱导的RAW 264.7细胞中产生一氧化氮(NO)具有抑制作用，因此本研究对二氯甲烷部位的提取物进行了系统的化学成分研究，确定其发挥抗炎药效的物质基础，从而发现新的抗炎活性先导化合物。

本研究采用多种分离纯化技术(正相硅胶、Sephadex LH-20凝胶、半制备高效液相色谱、薄层色谱、重结晶等)对新疆阿魏地上部位的二氯甲烷极性段进行了系统研究，从中共分离得到42个化合物，并依据其理化常数和光谱数据，结合单晶X射线衍射(X-Ray)和Mosher反应技术对其结构进行了鉴定，最终鉴定出36个香豆素及其衍生物类化合物，包括3个香豆素类化合物(FSA-1~ FSA-3)，1个单萜香豆素类化合物(FSA-4)，32个倍半萜香豆素类化合物(FSA-5~FSA-36)；此外，还鉴定了1个苯丙素类化合物(FSA-37)，1个木脂素类化合物(FSA-38)，1个吲哚类化合物(FSA-39)，3个类固醇类化合物(FSA-40~FSA-42)。其中10个为新化合物，结构全部为倍半萜香豆素类化合物，分别为：新疆阿魏素A (Fesinkin A) (FSA-5)、新疆阿魏素B (Fesinkin B) (FSA-6)、新疆阿魏素C (Fesinkin C) (FSA-8)、4′E-新疆阿魏素D (4′E-Fesinkin D) (FSA-9)、4′Z-新疆阿魏素D (4′Z-Fesinkin D) (FSA-10)、(4′R,5′S,6′R,9′R)-新疆阿魏醇A ((4′R,5′S,6′R,9′R)-Ferusingenol A) (FSA-14)、(4′S,5′R,6′R,9′R)-新疆阿魏醇A ((4′S,5′R,6′R,9′R)-Ferusingenol A) (FSA-15)、新疆阿魏素E (Fesinkin E) (FSA-23)、新疆阿魏素F (Fesinkin F) (FSA-24)、新疆阿魏素G (Fesinkin G) (FSA-30)。3个化合物(FSA-39、FSA-41、FSA-42)为首次从该属中分离得到。

所有分离得到的单体化合物均进行了体外抗炎活性筛选，结果显示，倍半萜香豆素类化合物对LPS诱导的RAW 264.7细胞产生的NO呈现出不同程度的抑制作用，因此推测该类化合物可能为新疆阿魏地上部位发挥抗炎活性的主要物质基础，尤其是化合物Episamarcandin acetate (FSA-31)和 Samarcandone (FSA-33)对NO释放量的抑制作用最为明显，IC₅₀值分别为2.3±0.1 μM和3.7±0.3 μM, 并且对正常的RAW 264.7细胞毒性较低。此外，我们还采取ELISA试剂盒检测了化合物FSA-31和FSA-33对RAW 264.7中炎症细胞因子白细胞介素-1β (IL-1β)、白细胞介素-6 (IL-6)和肿瘤坏死因子-α (TNF-α)产生的影响，结果显示化合物FSA-31在6.25 μM浓度下对IL-6释放量的抑制作用最为显著。最后，为了进一步研究倍半萜香豆素类化合物产生抗炎活性的作用靶点，我们选取化合物Episamarcandin acetate分别与iNOs和IL-6蛋白受体进行了分子对接模拟，此结果为发现新型抗炎药物提供了科学依据和理论基础。

同时，本文还对近二十年阿魏属植物中的倍半萜香豆素类化合物的研究情况进行了综述，为接下来的研究奠定基础。

Ferula sinkiangensis K. M. Shen belongs to the family Apiaceae, mainly distributed in Yili, Xinjiang Uygur Autonomous Region in China, which possesses various medicinal efficacy such as eliminating accumulation, removing abdominal mass, dispersing masses and killing insects. It is also a traditional medicine in China. However, due to the changes in the natural environment, long-term indiscriminate digging and other aspects of the restrictions, the growth environment of F. sinkiangensis has been greatly damaged, resulting in the increasing shortage of F. sinkiangensis resources. F. sinkiangensis has been listed as national secondary endangered plants in China. At present, only the resin of F. sinkiangensis and F. fukangensis is recorded in the 2020 edition of Chinese Pharmacopoeia as the medicinal part, and can only be produced by cutting roots and stems, which is very difficult to obtain. However, the aerial parts of F. sinkiangensis with many years of growth and large quantity are rarely used. In this research, the dichloromethane extracts which obtained from the aerial parts of F. sinkiangensis showed NO production inhibitory effects on LPS-induced RAW 264.7 cells, and its chemical constituents were systematically studied in order to discover the material basis for its anti-inflammatory effect. 

The dichloromethane extract from the aerial parts of F. sinkiangensis was systematically investigated by varies separation and purification methods such as silica gel, Sephadex LH-20 gel, semipreparative high performance liquid chromatography, thin layer chromatography (TLC) and recrystallization, and 42 compounds were isolated. Their structures were elucidated by spectral analysis, single crystal X-ray diffraction (X-Ray) and Mosher reaction. As a result, 36 coumarins and their derivatives were identified, including 3 coumarins (FSA-1~FSA-3), 1 monoterpene coumarin (FSA-4) and 32 sesquiterpene coumarins (FSA-5~FSA-36); in addition, 1 phenylpropanoid compound (FSA-37), 1 lignan compound (FSA-38), 1 indole compound (FSA-39) and 3 steroid compounds (FSA-40~FSA-42) were also identified. Among them, 10 compounds were new compounds, all of which were sesquiterpene coumarins: Fesinkin A (FSA-5), Fesinkin B (FSA-6), Fesinkin C (FSA-8), 4′E-Fesinkin D (FSA-9), 4′Z-Fesinkin D (FSA-10), (4′R,5′S,6′R,9′R)-Ferusingenol A (FSA-14), (4′S,5′R,6′R,9′R)-Ferusingenol A (FSA-15), Fesinkin E (FSA-23), Fesinkin F (FSA-24), Fesinkin G (FSA-30). Beyond that, 3 compounds (FSA-39、FSA-41、FSA-42) were firstly isolated from this genus.

All the purified compounds were tested in vitro anti-inflammatory activities on LPS-induced RAW 264.7 cells. The results showed that sesquiterpene coumarins had different degrees of inhibitory effects on the NO production, suggesting that these compounds may be the main material basis for the anti-inflammatory effect of the aerial parts of F. sinkiangensis. In particular, Episamarcandin acetate (FSA-31) and Samarcandone (FSA-33) showed significant inhibitory effects on the NO production (IC₅₀=2.3 ± 0.1 and 3.7 ± 0.3 μM, respectively) with lower toxicity against normal RAW 264.7 cells. Furthermore, we used ELISA kit to detect the effects of compounds FSA-31 and FSA-33 on the production of inflammatory cytokines IL-1β, IL-6 and TNF-α in LPS-induced RAW 264.7. The results showed that compound FSA-31 had the most significant inhibitory effect on IL-6 release under the concentration of 6.25 μM. Finally, molecular docking studies were carried out to predict the anti-inflammatory targets of natural sesquiterpene coumarins, compound Episamarcandin acetate was selected to perform molecular docking simulations with iNOs and IL-6 receptors, respectively, which provided a scientific and theoretical basis for the development of new anti-inflammatory drugs.

Besides, research advances on the sesquiterpene coumarins from Ferula genus in the past two decades were summarized, which may provide a reference for further research.