曼氏迭宫绦虫裂头蚴生长发育中糖代谢规律研究

Sparganosis is a parasitic zoonosis, and has seriously threatened human health for a long time. Presently, the treatment of sparganosis is mainly using surgery to remove the parasites, and there is no specific drugs used to control the sparganosis. Nowadays, cerebral sparganosis, proliferative sparganosis and subcutaneous sparganosis have no effective treatment. Drugs used in the clinical treatment of parasites mainly block parasites to obtain glucose and produce energy to kill them. Moreover, little study involving glycometabolism of sparganum has been done. In this study, we focus on building gene library of key enzymes involving glycometabolism, and detecting contents of glucogenic amino acid, glycerol, glucose and glycogen in the infected and uninfected frog muscle by high performance liquid chromatography (HPLC). Sparganum are cultivated with different nutrients mentioned above and inhibitors of key enzymes in vitro. This study explores the glycometabolism pathways which provide energy for sparganum survival through combining with HPLC, real time PCR, western blot, key enzymes activity detections, nucleic acid in situ hybridization and immunohistochemical localization analysis. This project carries out research on glycometabolism of sparganum to study the significant metabolic pathways and key enzymes in the glycometabolism, as well as to obtain drug target. It provides important theoretical basis for the follow-up drug development.

曼氏迭宫绦虫裂头蚴病是一种人畜共患寄生虫病，已严重威胁公共健康安全。目前临床上主要通过外科手术摘除虫体，尚无特效药物用于治疗裂头蚴病，而对于脑部裂头蚴病、增值型裂头蚴病和皮下多发型裂头蚴病更是束手无策。现阶段治疗寄生虫病的药物多数是通过阻断寄生虫对葡萄糖摄取和能量供应达到杀灭虫体，然而关于裂头蚴的糖代谢研究尚属空白。本研究拟通过建立糖代谢途径关键酶的基因组文库，利用高效液相色谱比较感染和未感染裂头蚴青蛙肌肉中生糖氨基酸、甘油、葡萄糖、糖原的含量。利用以上不同营养物和关键调节酶特异性抑制剂进行体外裂头蚴培养，联合高效液相色谱法、real-time PCR、western blot、关键调节酶活性检测、核酸原位杂交技术和免疫组织定位探究裂头蚴糖代谢途径之间如何协同作用提供能量。本项目将揭示裂头蚴的糖代谢规律，从而获取关键药物靶点为后续的药物开发提供重要的理论依据。

曼氏迭宫绦虫裂头蚴病是一种人畜共患寄生虫病，已严重威胁公共健康安全。目前临床上主要通过外科手术摘除虫体，尚无特效药物用于治疗裂头蚴病，而对于脑部裂头蚴病、增值型裂头蚴病和皮下多发型裂头蚴病更是束手无策。现阶段治疗寄生虫病的药物多数是通过阻断寄生虫对葡萄糖摄取和能量供应达到杀灭虫体。糖类（葡萄糖和糖原）被认为是蠕虫重要的能量来源，糖酵解是获取能量的主要方式，少数绦虫也可以通过皮层吸收宿主体内的氨基酸、脂肪酸、甘油等作为营养来源。本课题研究发现，感染青蛙肌肉组织中检测到生糖氨基酸、糖原和葡萄比正常青蛙肌肉组织中的高，且存在显著性差异，只有甘油成分在两者中均没有检测到，说明生糖氨基酸、糖原和葡萄糖是裂头蚴的主要营养来源，利用这三种营养物质进行体外裂头蚴培养，发现裂头蚴的生长情况与培养物浓度在一定程度上呈现正向关联。同时构建了糖代谢途径的代谢酶基因组文库，在裂头蚴阶段五种糖代谢途径是完整的，但是通过检测限速酶转录水平发现，三羧酸循环、糖酵解、糖异生和糖原分解途径的转录水平均较高，生糖氨基酸在代谢过程中经过一系列转变为磷酸烯醇式丙酮酸最后经过糖异生途径合成葡萄糖；糖原在糖原磷酸化酶的作用下水解出葡萄糖，裂头蚴可通过三羧酸循环和糖酵解途径利用葡萄糖产生能量供应，从而推断出这些途径在裂头蚴生长发育过程中发挥了能量供应协同作用，而磷酸戊糖途径和糖原合成途径中的限速酶转录水平较低，可能在能量供应中发挥作用不大。本课题研究揭示了三羧酸循环、糖酵解、糖异生和糖原分解途径在裂头蚴生长发育中发挥重要作用，其对应的限速酶具有潜在开发价值的药物靶标。