中国大鲵小肠代谢相关蛋白的表达及预示的代谢活性分析

发布时间：2018-06-03 01:41

本文选题：中国大鲵 + 小肠　；参考：《河南师范大学》2017年硕士论文

【摘要】：中国大鲵(Andrias davidianus)又称娃娃鱼,隶属两栖纲、有尾目、隐鳃鲵科,是世界上现存个体最大的有尾目两栖类。大鲵有“活化石”之称,是我国的II级保护动物。近年来受多种因素的影响,中国大鲵的分布范围逐渐缩小,资源受到极大的破坏,对大鲵的保护刻不容缓。中国大鲵属于由水生向陆生转变的过渡两栖动物,且其迁移捕食能力较差,是研究生物进化和多样性的好材料。大鲵小肠作为重要的消化和吸收器官,在超微结构上已经具备了较强的消化、吸收、合成和分泌功能的结构基础,如细胞质中线粒体、内质网、高尔基体和溶酶体丰富。目前中国大鲵的研究主要集中在组织形态学,但在分子层面的研究未见报道。因此,本文采用iTRAQ标记结合LC-MS/MS的蛋白质研究技术对正常中国大鲵的小肠进行蛋白检测。然后用GO、NCBI、GENMAPP、KEGG、QIAGEN和IPA软件等生物信息学方法对鉴定到的蛋白进行生理活动分类,挑选出糖代谢、脂代谢、蛋白质和氨基酸代谢以及核酸代谢相关蛋白,之后用IPA软件这些蛋白进行通路分析。最后通过查阅相关文献资料,研究代谢相关信号通路中的主要蛋白在中国大鲵小肠中的功能作用。主要结果如下:(1)参与中国大鲵小肠糖类代谢的蛋白有117种。涉及糖代谢的有:糖酵解和糖异生途径,主要蛋白有ALOA、ALOB、ENO1、ENO2、ENO3和GPI等;磷酸戊糖途径,主要蛋白有PGD和PGLS等。(2)参与中国大鲵小肠脂代谢的蛋白有142种,涉及脂代谢的有:脂肪酸β氧化途径,主要蛋白有ACADM、ACSL1、ECHS1、HADHA和HADHB等;酮体生成途径,主要蛋白有BDH1和HMGCS1等。在中国大鲵小肠糖代谢和脂代谢中均检测到了AMPK信号通路,主要蛋白为CYP7B1。(3)参与中国大鲵小肠蛋白质及氨基酸代谢的蛋白有121种,主要涉及蛋白泛素化途径,主要蛋白有PSMC1、PSMC2、PSMC3、PSMC4和PSMC5等;AHR信号通路,主要涉及的蛋白有SLC7A8和SLC3A1等;组胺降解途径,主要蛋白有DAO等。(4)参与在中国大鲵小肠核酸代谢的蛋白有105种,涉及嘧啶核苷酸从头合成途径的主要蛋白有AK1、NME7、NUDT9和UMPS等;SAPK/JNK信号通路,主要蛋白有CDC42,CRK和CRKL等。综上所述,本文用iTRAQ标记结合LC-MS/MS的蛋白质研究技术检测了中国大鲵正常小肠中参与糖代谢、脂代谢、蛋白质和氨基酸代谢以及核酸代谢相关蛋白的表达丰度,用生物信息学方法分析了中国大鲵小肠中代谢活动及其涉及的信号通路,通过查阅相关文献查找出在中国大鲵正常小肠中可能参与调节代谢相关信号通路的主要蛋白,为进一步研究中国大鲵小肠的生理功能提供资料。
[Abstract]:The Chinese giant salamander Andrias davidianus, also called the baby fish, belongs to the amphibian class, which has the order Carnionidae, which is the largest living amphibian species in the world. Giant salamander, known as a "living fossil", is a class II protected animal in China. In recent years, due to the influence of many factors, the distribution of Chinese giant salamander has been gradually reduced, and the resources have been greatly damaged, so it is urgent to protect the giant salamander. Chinese giant salamander belongs to the transition amphibian from aquatic to terrestrial, and its migration and predation ability is poor, so it is a good material to study biological evolution and diversity. As an important digestive and absorptive organ, the small intestine of giant salamander has strong structural basis of digestion, absorption, synthesis and secretion in ultrastructure, such as mitochondria, endoplasmic reticulum, Golgi body and lysosome in cytoplasm. At present, the studies of Chinese giant salamander mainly focus on histomorphology, but there is no report on molecular level. Therefore, iTRAQ labeling combined with LC-MS/MS protein technique was used to detect the protein in the small intestine of Chinese giant salamander. Then the identified proteins were classified by bioinformatics methods, such as GONCBI- GENMAPPP-KEGGN and IPA software. The proteins related to glucose metabolism, lipid metabolism, protein and amino acid metabolism and nucleic acid metabolism were selected. IPA software was used to analyze the pathway of these proteins. Finally, the function of major proteins in metabolic signaling pathway in the small intestine of Chinese giant salamander was studied. The main results are as follows: 1) 117 kinds of proteins are involved in saccharide metabolism in the small intestine of Chinese giant salamander. Glycolysis and glycometabolism are involved in glycolysis and glycometabolism, the main proteins are AlOAA, ENO1, ENO2, ENO3 and GPI, and the pentose phosphate pathway, which mainly involves PGD and PGLS, has 142 kinds of proteins involved in lipid metabolism in the small intestine of Chinese giant salamander (Andrias davidianus). Fatty acid 尾 oxidation pathway was involved in lipid metabolism, the main proteins were ACADML ACSL 1 ECHS1 HADHA and HADHB, and ketone body formation pathway, BDH1 and HMGCS1, etc. AMPK signaling pathway was detected in the small intestine glucose and lipid metabolism of Chinese giant salamander. The main protein was CYP7B1.3) there were 121 proteins involved in the metabolism of protein and amino acids in the small intestine of Chinese giant salamander, mainly involved in the ubiquitin pathway. The main proteins include PSMC1, PSMC2, PSMC3, PSMC4 and PSMC5, mainly involved in SLC7A8 and SLC3A1, histamine degradation pathway, DAO et al. 4) there are 105 kinds of proteins involved in nucleic acid metabolism in the small intestine of Chinese giant salamander. The major proteins involved in the ab initio synthesis of pyrimidine nucleotides are AK1NME7NUDT9 and UMPS. The major proteins are CDC42 CRK and CRKL. In conclusion, iTRAQ labeling combined with LC-MS/MS protein technique was used to detect the abundance of proteins involved in glucose metabolism, lipid metabolism, protein and amino acid metabolism and nucleic acid metabolism in the normal small intestine of Chinese giant salamander. The metabolic activity in the small intestine of Chinese giant salamander and its signal pathway were analyzed by bioinformatics. The main proteins involved in the regulation of metabolic signaling pathway in the normal small intestine of Chinese giant salamander were found by consulting the relevant literature. To further study the physiological function of the small intestine of Chinese giant salamander.
【学位授予单位】：河南师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S917.4

【参考文献】