岩藻糖化糖胺聚糖的寡糖结构及其抗乙酰肝素酶活性研究

	岩藻糖化糖胺聚糖的寡糖结构及其抗乙酰肝素酶活性研究; Study on the chemical structure of oligosaccharides from fucosylated glycosaminoglycan and their heparanase inhibitory activities
	周路坦
导师	赵金华
摘要	Invasion and metastasis of tumor cells is an important feature of malignant tumors and also the major obstacle to its clinical treatment. Extracellular matrix (ECM) and basement membrane (BM) serve as the main histological barriers for cell metastasis and angiogenesis. Heparan sulfate (HS) contained in them can be blocked by heparanase (HPSE) degradation. Studies have found that inhibiting or blocking HPSE activity can inhibit tumor cell invasion and metastasis. Further studies have confirmed that HPSE is closely related to angiogenesis and tumor invasion and metastasis. Additionally, venous thromboembolism (VTE) is a common malignant tumor complication. In theory, drugs with both anti-tumor and anti-thrombotic activities are beneficial to tumor treatment. Heparin (Hep), as a classic anticoagulant, is a common drug for clinical treatment of malignant tumor thromboembolism complications. However, the clinical application of Hep is severely limited due to the serious risk of bleeding. Fucosylated glycosaminoglycan (FG) extracted from sea cucumber is a glycosaminoglycan derivative fucose branches. Depolymerized FG and its purified fragments (oligosaccharides) showed potent anticoagulant and antithrombotic activities with low bleeding tendency through selectively inhibiting intrinsic factor Xase (iXase). Given that FG has a similar but different backbone structure of glycosaminoglycans from HS, it may have heparin-related activities. Purpose: This dissertation explores the effect of purified oligosaccharides from FG (oFGs) on HPSE activity, and then further analyzed the effects of oFGs on angiogenesis and tumor cell metastasis. The purpose was to discover active ingredients that both have anti-tumor and anti-thrombotic effects with potential clinical applications and without serious bleeding risk. Methods: (1) Using β-elimination combined with gel column chromatography method to separate and purify series of oligosaccharides (oFGs). Then using NMR analysis combined with mass spectrometry data to confirm the structure of the oligosaccharides. (2) Homogeneous time resolved fluorescence resonance energy transfer (HTRF) method was used to analyze the effect of FGs on HPSE activity, and surface plasmon resonance (SPR) method and computer simulation technology were used to analyze its interaction with target protein mechanism. (3) Cell culture in vitro combined with Transwell chamber and Matrigel were used to analyze the effects of oFGs on the migration and invasion of tumor cells and vascular endothelial cells. (4) Matrigel tubule formation and chick embryo chorioallantoic models were studied the effects of oFGs on angiogenesis in vitro and in vivo, respectively. (5) Animal experimental model was used to study the effects of oFGs on tumor cell metastasis and colonization in mice. Results: (1) Preparation and structure analysis of oFGs: The native fucosylated glycosaminoglycan isolated from sea cucumber Holothuria fuscopunctata (nHG) was depolymerized by a
	2020-11
文献类型	学位论文
条目标识符	http://ir.kib.ac.cn/handle/151853/74232
专题	昆明植物所硕博研究生毕业学位论文
推荐引用方式 GB/T 7714	周路坦. 岩藻糖化糖胺聚糖的寡糖结构及其抗乙酰肝素酶活性研究, Study on the chemical structure of oligosaccharides from fucosylated glycosaminoglycan and their heparanase inhibitory activities[D],2020.

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