本文選題：中國大鯢 + 小腸　； 參考：《河南師范大學(xué)》2017年碩士論文

【摘要】：中國大鯢(Andrias davidianus)又稱娃娃魚,隸屬兩棲綱、有尾目、隱鰓鯢科,是世界上現(xiàn)存?zhèn)�體最大的有尾目兩棲類。大鯢有“活化石”之稱,是我國的II級保護動物。近年來受多種因素的影響,中國大鯢的分布范圍逐漸縮小,資源受到極大的破壞,對大鯢的保護刻不容緩。中國大鯢屬于由水生向陸生轉(zhuǎn)變的過渡兩棲動物,且其遷移捕食能力較差,是研究生物進化和多樣性的好材料。大鯢小腸作為重要的消化和吸收器官,在超微結(jié)構(gòu)上已經(jīng)具備了較強的消化、吸收、合成和分泌功能的結(jié)構(gòu)基礎(chǔ),如細胞質(zhì)中線粒體、內(nèi)質(zhì)網(wǎng)、高爾基體和溶酶體豐富。目前中國大鯢的研究主要集中在組織形態(tài)學(xué),但在分子層面的研究未見報道。因此,本文采用iTRAQ標記結(jié)合LC-MS/MS的蛋白質(zhì)研究技術(shù)對正常中國大鯢的小腸進行蛋白檢測。然后用GO、NCBI、GENMAPP、KEGG、QIAGEN和IPA軟件等生物信息學(xué)方法對鑒定到的蛋白進行生理活動分類,挑選出糖代謝、脂代謝、蛋白質(zhì)和氨基酸代謝以及核酸代謝相關(guān)蛋白,之后用IPA軟件這些蛋白進行通路分析。最后通過查閱相關(guān)文獻資料,研究代謝相關(guān)信號通路中的主要蛋白在中國大鯢小腸中的功能作用。主要結(jié)果如下:(1)參與中國大鯢小腸糖類代謝的蛋白有117種。涉及糖代謝的有:糖酵解和糖異生途徑,主要蛋白有ALOA、ALOB、ENO1、ENO2、ENO3和GPI等;磷酸戊糖途徑,主要蛋白有PGD和PGLS等。(2)參與中國大鯢小腸脂代謝的蛋白有142種,涉及脂代謝的有:脂肪酸β氧化途徑,主要蛋白有ACADM、ACSL1、ECHS1、HADHA和HADHB等;酮體生成途徑,主要蛋白有BDH1和HMGCS1等。在中國大鯢小腸糖代謝和脂代謝中均檢測到了AMPK信號通路,主要蛋白為CYP7B1。(3)參與中國大鯢小腸蛋白質(zhì)及氨基酸代謝的蛋白有121種,主要涉及蛋白泛素化途徑,主要蛋白有PSMC1、PSMC2、PSMC3、PSMC4和PSMC5等;AHR信號通路,主要涉及的蛋白有SLC7A8和SLC3A1等;組胺降解途徑,主要蛋白有DAO等。(4)參與在中國大鯢小腸核酸代謝的蛋白有105種,涉及嘧啶核苷酸從頭合成途徑的主要蛋白有AK1、NME7、NUDT9和UMPS等;SAPK/JNK信號通路,主要蛋白有CDC42,CRK和CRKL等。綜上所述,本文用iTRAQ標記結(jié)合LC-MS/MS的蛋白質(zhì)研究技術(shù)檢測了中國大鯢正常小腸中參與糖代謝、脂代謝、蛋白質(zhì)和氨基酸代謝以及核酸代謝相關(guān)蛋白的表達豐度,用生物信息學(xué)方法分析了中國大鯢小腸中代謝活動及其涉及的信號通路,通過查閱相關(guān)文獻查找出在中國大鯢正常小腸中可能參與調(diào)節(jié)代謝相關(guān)信號通路的主要蛋白,為進一步研究中國大鯢小腸的生理功能提供資料。
[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.
【學(xué)位授予單位】：河南師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S917.4

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