羧酸酯酶-1作为早期肝损伤新型标志物的初步研究

发布时间：2018-04-03 21:53

本文选题：羧酸酯酶1　切入点：肝损伤　出处：《大连医科大学》2017年硕士论文

【摘要】：哺乳动物的羧酸酯酶(CEs)是重要的丝氨酸水解酶家族的成员,Ces可以分为5个亚型,但最主要的亚型是羧酸酯酶1(CE1)和羧酸酯酶2(CE2),主要存在于组织的内质网腔内。人类羧酸酯酶1(Human Carboxylesterases 1,hCE1)作为一类代谢酶,广泛存在于人体肝脏、肾脏及皮肤等组织中,但却在肝脏中特异性分布且含量极高。hCE1负责水解各种内源性物质包括胆固醇酯,甘油三酯等,并在维持机体脂质代谢平衡中起着关键的作用。此外还催化水解各种外源性物质,如某些临床药物和环境毒素,在机体的防御体系中也扮演重要角色。由于hCE1在肝脏中特异性分布,当肝脏收到损伤时,肝细胞破裂后释放入血,在血中可以检测到该酶。肝脏为人体极为重要的脏器之一,是体内各种碳水化合物、蛋白质、脂质、维生素合成代谢的主要器官,并且在分泌、排泄、解毒、生物转化等方面也发挥着重要作用。基于肝特殊的解剖位置及其生理功能,多种内源性或外源性的因素,比如:病毒、细菌、炎症因子、药物、环境毒物等都能引起肝功能不同程度受损,最终导致疾病发生。肝损伤的准确预测和诊断是临床医学的研究热点,受到研究者的广泛关注。由于损伤的机制较为复杂,影响因素多,常常涉及到诸多代谢反映的参与,或是引起免疫反应,因此其检查方法依然存在很多问题。较好的血清标志物需要具有快速,简便,灵敏准确且早期就能反应出问题所在。目前临床上对肝损伤主要是进行肝生化检查,包括一些肝脏酶学检查,肝合成功能及肝纤维化血清学指标等,能较全面的反映肝功能。其中肝纤维化指标较为特异,但不够敏感。考虑到这些指标不能有效进行分类,不能较早的反映肝功能,需要结合其他的相关指标进行综合考虑。随着近年来蛋白组学、代谢组学和临床检验相关学科和技术的快速发展,越来越多的新型血清学指标将被发现,弥补了传统指标的一些不足,但是大部分尚停留在研究阶段。因此,发现并验证新型血清标志物对于肝损伤的早期诊断与治疗具有重要意义。本文采用羧酸酯酶1的特异性小分子荧光探针对血浆中hCE1的进行检测,该探针为hCE1的底物,被hCE1代谢后产生相应的产物,通过高效液相色谱荧光分析,根据产物的生成量,以测定羧酸酯酶的活性。用此方法检测不同原因造成的肝损伤血清中的hCE1的活性,以此来分析探求该酶与不同肝脏疾病的关系,并与目前临床比较认可的肝损伤标准进行相关性分析,探究其作为新型标志物的可行性及意义。
[Abstract]:Ces, a member of the important serine hydrolase family, can be divided into five subtypes, but the main subtypes are carboxylesterase 1 (CE1) and carboxylesterase 2 (CE2), which mainly exist in the endoplasmic reticulum (ER) cavity of tissues.Human carboxylesterase 1(Human Carboxylesterases 1hCE1 is a kind of metabolic enzyme, which widely exists in human liver, kidney, skin and other tissues. However, it is specifically distributed in the liver and contains a high content of HCE1, which is responsible for the hydrolysis of various endogenous substances, including cholesterol ester.Triglyceride plays a key role in maintaining the balance of lipid metabolism.In addition, the catalytic hydrolysis of various exogenous substances, such as some clinical drugs and environmental toxins, also plays an important role in the body's defense system.Because of the specific distribution of hCE1 in the liver, when the liver is damaged, the liver cells release into the blood after rupture, and the enzyme can be detected in the blood.As one of the most important organs in human body, liver is the main organ of carbohydrate, protein, lipid, vitamin synthesis and metabolism, and also plays an important role in secretion, excretion, detoxification, biotransformation and so on.Based on the special anatomical location and physiological function of the liver, many endogenous and exogenous factors, such as virus, bacteria, inflammatory factors, drugs and environmental poisons, can damage liver function to varying degrees and eventually lead to disease.The accurate prediction and diagnosis of liver injury is a hot topic in clinical medicine.Because the mechanism of injury is more complex and there are many factors affecting it, it often involves many metabolic reactions, or causes immune response, so there are still many problems in its examination method.Better serum markers need to be quick, simple, sensitive, accurate and early to respond to the problem.At present, liver injury is mainly examined by biochemical examination of liver, including some liver enzyme examination, liver synthesis function and liver fibrosis serological index, which can reflect liver function comprehensively.The indexes of liver fibrosis were specific, but not sensitive.Considering that these indexes can not be classified effectively and can not reflect liver function earlier, it needs to be considered synthetically with other related indexes.With the rapid development of proteomics, metabolomics and clinical laboratory related disciplines and techniques, more and more new serological indicators will be found, which make up for some shortcomings of traditional indicators, but most of them are still in the research stage.Therefore, it is important to find and verify new serum markers for early diagnosis and treatment of liver injury.In this paper, hCE1 in plasma was detected by using a specific small molecular fluorescence probe of carboxylesterase 1. The probe was the substrate of hCE1, which was metabolized by hCE1 to produce corresponding products. The products were analyzed by high performance liquid chromatography (HPLC).To determine the activity of carboxylate esterase.This method was used to detect the activity of hCE1 in the serum of liver injury caused by different causes, so as to analyze the relationship between the enzyme and different liver diseases, and to analyze the correlation between the enzyme and the current approved liver injury standard.Explore its feasibility and significance as a new type of marker.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R575

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