【作者】 刘莎；

【导师】 蔡柏蔷； 许文兵；

【作者基本信息】 北京协和医学院 ， 临床医学， 2010， 博士

【摘要】 特发性肺纤维化(IPF)是一种有着较高病死率的疾病,其特征为肺部成纤维细胞的聚集、大量胶原沉积以及基质重塑。目前人们认为肺泡上皮细胞的上皮-间质转化(epithelial to mesenchymal transition, EMT)这一过程可能在肺纤维化的发病过程中发挥了重要作用,并且TGF-β1可以促进这一过程的发生。糖胺聚糖类药物在临床多用于抗凝,近来越来越多的证据证明以肝素为代表的糖胺聚糖类药物对于器官纤维化有抑制作用。有研究表明其抗纤维化作用可能是通过其对凝血系统激活的抑制产生的,但还有一些研究者发现糖胺聚糖类药物本身也有抗纤维化作用,它们可以抑制TGF-β1在纤维化组织中的过度表达。舒洛地特是从猪小肠粘膜提取的天然物质,由四种糖胺聚糖组成。它与低分子肝素相比有着更少的副作用和更方便的用药方式。目前尚无其对于肺纤维化作用的报道。本研究以体外培养肺上皮A549细胞为基础,通过细胞形态学观察TRF-PCR法以及免疫荧光法检测舒洛地特对于TGF-β1诱导的EMT过程相关标志物表达的影响,来验证舒洛地特在抗凝作用之外的抗纤维化作用。结果显示,舒洛地特能够部分抑制TGF-β1诱导的肺泡上皮细胞形态的变化和间质细胞标志物基因水平和蛋白水平的表达,说明它本身对EMT过程有抑制作用,这也为IPF的治疗提供了一种新的研究方向。更多还原

【Abstract】 Idiopathic pulmonary fibrosis (IPF) is a disease characterized by fibroblast accumulation, excessive collagen deposition and matrix remodeling which continues to be associated with considerable morbidity and mortality. Now people think the process termed "epithelial-mesenchymal transition"(EMT) may play an important role in fibrogenesis and it has been proved that TGF-β1can induce this process. Nowadays there is compelling evidence that glycosaminoglycans, for example, heparin which are mainly used as anticoagulant can inhibit organ fibrosis. Some researchers found that this kind of medicine may exert their anti fibrotic effect by influencing the activated coagulation cascade. But some people think glycosaminoglycans have their own anti fibrotic effect which is not associated with the haemostatic system, for example, they found that glycosaminoglycans can prevent TGF-β1overexpression in fibrotic tissues. Sulodexide is isolated from porcine intestinal mucosa and comprised of four naturally glycosaminoglycan components. Compared to low-molecular heparin, it has less side effects and can be taken orally. The influence of sulodexide on pulmonary fibrosis has not been studied yet. This study which is based on the culture of human lung epithelial cells A549in vitro used morphology examination, RT-PCR, Immunofluorescence methods to evaluate the effect of Sulodexide on the marker expression in TGF-β1induced EMT process. So it can determine whether it has anti fibrotic activity beyond its anticoagulation effect. The results show the Sulodexide can partially inhibit the morphology changes and decrease the mesenchymal marker expression induced by TGF-β1. This indicate that sulodexide can inhibit the EMT process and it may present a potential novel anti-fibrotic therapeutic approach for the future.更多还原