内脏利什曼病小鼠模型的代谢组学研究及生物标志物筛选

Visceral Leishmaniasis (Kala-azar) is a kind of parasitic disease that can cause human death. In China, this disease is mainly endemic in northwest and border areas. Recent years, the prevalence and morbidity of leishmaniasis increased graduately. Previous phylogenic evidence that based on biomarkers showed that the primary pathogeny of Chinese Kala-azar is Leishmania Donovani. It is still unclear that the changes of metabolic level of hosts after the parasite’s invasion, and there is still no detecting technique that can diagnose Kala-azar accurately in its early phase. In this study project, the mouse model of visceral leishmaniasis wouled be established. The blood, urine, liver and spleen of infected mouse would be detected by metabolomics method that contain HPLS-MS and bioinformatics to analyze the changes of metabolic pathways that the different metabolites take part in, the metabolic level of hosts at different time points after infection, the relationship between metabolic level and the severity of leishmaniasis, and screen the potential metabolic biomarkers of visceral leishmaniasis by ROC-curve analysis as well. This study project will help us further understanding the mechanism of Leishmania-host interactions, and lay the foundation of developing early diagnosis techniques for Kala-azar.

黑热病是流行于我国西北部及边疆地区的一种可致人死亡的严重寄生虫病，且近年来有扩大流行和发病率逐渐增加的趋势。前期基于分子标记的种群鉴定结果证实我国黑热病病原体主要为杜氏利什曼原虫，而关于其感染后宿主的代谢水平的变化尚不清楚，且目前国内尚无一种能够准确早期诊断黑热病的实验室检测技术。本研究拟构建内脏利什曼病小鼠感染模型，采用代谢组学研究方法，以HPLC-MS技术及生物信息学分析手段，对感染杜氏利什曼原虫的小鼠的血液、尿液及肝脾组织进行非靶向代谢组学分析，寻找差异代谢物并分析其参与的机体代谢通路的变化，同时检测机体在感染后不同时期的代谢水平，分析代谢水平变化与内脏利什曼病发展程度之间的联系，同时通过ROC曲线分析筛选潜在的内脏利什曼病生物标志物。该研究为进一步认识利什曼原虫与宿主相互作用的机制及开发黑热病早期诊断技术奠定基础。

黑热病是一种可致人死亡的严重寄生虫病，现主要流行于我国西北部及边疆地区的，近年来有扩大流行和发病率逐渐增加的趋势。本项目在前期分子标记和种群鉴定研究的基础上，成功构建内脏利什曼病金黄地鼠模型，并利用代谢组学的研究策略，对感染黑热病的小鼠的血液、尿液样本进行了非靶向代谢组学分析，通过对感染后样本不同时间点的代谢水平进行分析，探究代谢变化与内脏利什曼病进展之间的联系，同时通过ROC曲线筛选潜在的诊断标志物。本项目研究发现在感染利什曼原虫后，鼠血清中甘油磷脂类代谢物受影响较大，进一步分析发现磷脂酰胆碱（Phosphatidylcholine）和磷脂酰乙醇胺（Phosphatidylethanolamine）在感染后出现显著增多，由此提示其在黑热病发病中的关键作用。对尿液样本的分析表明在感染早期糖类无氧代谢途径受影响较大，感染晚期氨基酸代谢受影响较大。此外，项目研究共确定16个潜在的可用于诊断的生物标志物，其中血清和尿液标志物各8个。研究为进一步认识利什曼原虫与宿主相互作用的机制及开发黑热病早期诊断技术奠定了基础。