【作者】 黎昌莉；

【导师】 蔡绍皙；

【作者基本信息】 重庆大学 ， 药学， 2010， 硕士

【摘要】 血管内皮细胞迁移在多种生理、病理现象中扮演着至关重要的角色。血管内皮细胞迁移有助于维持血管壁内皮的完整性和新血管的生长:在伤口愈合过程中,增殖的血管内皮细胞首先迁移到受损血管处,再以出芽或非出芽的方式形成新的血管;在组织修复的初始阶段,血管内皮细胞迁移到新生的肉芽组织中形成初始状态的毛细血管为新生的肉芽组织提供营养;另外,血管内皮细胞的迁移也是肿瘤的必要条件。研究能够诱导血管内皮迁移的中药对科研和临床有重要的意义,美洲大蠊提取液在临床中被广泛运用于皮肤创伤和胃溃疡治疗,且疗效明确。有大量的研究分别从促进胞基质表达,调节细胞因子分泌量,提高中性粒细胞数量,提高巨噬细胞吞噬能力及促使体内SOD值回升等角度解释美洲大蠊提取物促进伤口愈合的原因。但少有美洲大蠊提取物对血管内皮细胞作用的研究报道。因此,本研究通过传统的细胞迁移实验方法结合微流控芯片技术,对美洲大蠊提取液诱导血管内皮细胞迁移进行初步研究。对美洲大蠊虫体进行有效成分的提取;使用传统的细胞划痕法、Transwell小室法和琼脂糖平板法对不同浓度条件下的美洲大蠊提取液诱导血管内皮细胞迁移进行研究;运用插管抽丝法制作一款能够在较短时间内产生稳定浓度梯度的琼脂糖微流控装置,并借助该装置检测美洲大蠊提取液诱导血管内皮细胞迁移。运用划痕法和Transwell小室法初步证实美洲大蠊提取物能诱导血管内皮细胞迁移:其中5%(体积比)的美洲大蠊提取物对血管内皮的迁移没有发生明显的迁移作用,在浓度为5-30%范围内,血管内皮细胞的迁移呈在线性关系。利用琼脂糖凝胶为基质的微流控装置中以20%浓度的美洲大蠊提取物为诱导剂诱导6h后,将细胞培养去等分为4个区,其中1区和2区分别有大约7%和3%的细胞减少,而3区和4区分别有大约9%和2%的细胞数目增加。初步证明美洲大蠊提取物能够诱导血管内皮细胞迁移;同时插管抽丝法制作的琼脂糖微流控装置能够产生稳定的浓度梯度,可以开发成用于检测细胞迁移的装置。更多还原

【Abstract】 Endothelial cell migration play improtant role in many physiological and pathologic process.The integrity of vessel endothelium and angiogenesis depend on endothelial cell migration:in the process of wound healing,neovascularization by budding or other way is the consquence of the proliferous EC migrate to the breakage of vessel;in the elementary step of tissue repair,the capillary network in original state supplying granulation tissue with nutrientsubstance is the consquence of the proliferous EC migrate to anagenetic granulation tissue;additionally,the migration of EC is prerequisite for tumorigeness. Research on what chinese medicine induce EC migrate will be make contribution to clinic. The extraction from periplaneta americana were ofen used for treatment of wound healling and gastric ulcer effectively.There are a great deal of research on mechanism of periplaneta americana for wound healling,such as it adjust the secretory volume of EC,incerase the quantity of neutrophilic granulocyte, advance the endocytosis-ability of macrophage or promote the SOD level rally. But rerely research on what will happen to extraction of periplaneta americana act on EC. Therefore, in our reserch get help from tradional laboratory facilities and micro-fluidic chip to prove the chemotaxis of endothelial cells in response to the extraction from periplaneta americana.The solution is extract from periplaneta americana then using the traditional methods of scratch assay , transwell chamber and agarose plate test the extraction of periplaneta americana in various consistency induce EC migrate. A microfluidic device which using withdrawing thread form agarose hydrogel was designed and fabricated to generate stable concentration gradients immediately . Using this device monitored the chemotactic response of endothelial cell to the extraction of periplaneta americana.The methods of scratch assay and transwell chamber approved conformably that extraction of periplaneta americana can induce EC migrate: the extraction of medium and periplaneta americana volume ratio at 5% were unable to induce human endothelial cells migration. In contrast, to the extent that volume ratio is 10% to 30%, the relation of extraction concentration and the quantity of EC migration is linear dependence. There are respectively 7% and 3% of human endothelial cells were observed to migrate towards higher concentrations when the Medium and Periplaneta americana volume ratio was 20%.更多还原