基于抑制结肠癌血管新生的准噶尔阿魏物质基础及作用机理研究

阿魏是新疆特色药材，前期研究表明“香阿魏”(准噶尔阿魏、多伞阿魏)抗结肠癌强于“臭阿魏”(新疆阿魏、托里阿魏)，“臭阿魏”濒临灭绝，准噶尔阿魏资源丰富、可抑制斑马鱼血管新生，国内外未见报道。本课题采用硅胶柱色谱对准噶尔阿魏提取分离，以绿色荧光血管转基因斑马鱼血管新生为指标，进行活性跟踪，筛选活性部位、化合物，通过UV、IR、HPLC-MS、GC-MS、NMR进行结构解析；一是体外实验，检测准噶尔阿魏活性部位、化合物对结肠癌细胞HCT 116划痕、侵袭、迁移，血管内皮细胞管形成的影响；二是体内实验，建立裸鼠结肠癌原位种植模型，测定抑瘤率、肿瘤转移、肝肾功能，免疫组化检测微血管密度(MVD)、VEFGA、Dll4、Notch4；最后，Western blot测定准噶尔阿魏调控Dll4-Notch/VEGF信号开关抑制结肠癌血管新生的机制。旨在带动新疆18种“香阿魏”研究，促进民族医药产业发展。

Ferula is a characteristic medicine of Xinjiang,Previous studies have shown that the anticolonal activity of “Sweet Ferula” (Ferula soongarica,Ferula feurlaeoides) is stronger than that of “Smelly Ferula” (Ferula krylovii,Ferula sinkiangensis), “Smelly Ferula”is endangered,Ferula soongarica is rich in resources and can inhibit angiogenesis in zebrafish.It is not reported at China and abroad.In this study, Silica gel column chromatography was used for extraction and separation of Ferula soongarica, and angiogenesis of green fluorescent vascular transgenic zebrafish was used as the index to conduct activity tracking and screening of active sites and compounds.The structure was analyzed by UV、IR、HPLC-MS、GC-MS、NMR.Firstly, in vitro experiments were carried out to detect the effects of active sites and compounds of Ferula soongarica on the scratch, invasion and migration of colon cancer cells HCT 116 and the formation of vascular endothelial cell tubes. Secondly, in vivo experiments were conducted to establish an in situ implantation model of colon cancer in nude mice, to determine tumor inhibition rate, tumor metastasis, liver and kidney function, and to detect microvascular density (MVD), VEFGA, Dll4, and Notch4 by immunohisto chemistry. Finally, Western blot was used to determine the mechanism of regulation of Dll4-Notch/VEGF by Ferula soongarica in inhibiting angiogenesis in colon cancer. It aims to promote the research of 18 kinds of“Sweet Ferula” in Xinjiang and promote the development of ethnic medicine industry.