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高糖高脂日糧對(duì)團(tuán)頭魴生長(zhǎng)性狀、肝組織結(jié)構(gòu)及轉(zhuǎn)錄組的影響

發(fā)布時(shí)間:2021-11-01 23:31
  團(tuán)頭魴是中國(guó)重要的淡水草食性混養(yǎng)魚(yú)類之一。近年來(lái),如何最大限度地利用非蛋白能量飼料及其在飼料中如何正確配比等問(wèn)題已有較多研究與報(bào)道。非蛋白能量飼料的過(guò)量使用可以造成魚(yú)類體脂的大量沉積,從而導(dǎo)致魚(yú)類脂肪肝疾病。然而迄今為止,有關(guān)非蛋白能量飼料過(guò)量使用導(dǎo)致團(tuán)頭魴脂肪肝疾病發(fā)生的相關(guān)報(bào)道較少。通常情況下,魚(yú)類攝食是由餌料能量及營(yíng)養(yǎng)成分等因素協(xié)同調(diào)節(jié)影響魚(yú)類健康,而魚(yú)類健康又與相關(guān)功能基因的表達(dá)有關(guān),類似的研究已在其它魚(yú)類中獲得證實(shí),如虹鱒、金鯧等。團(tuán)頭魴該方面的相關(guān)研究,主要集中在碳水化合物與脂肪飼料對(duì)團(tuán)頭魴生長(zhǎng)性能及免疫機(jī)制改善等方面的研究,而由于日糧中高糖高脂導(dǎo)致團(tuán)頭魴脂肪沉積所產(chǎn)生的生長(zhǎng)性能和肝組織改變,以及其變化后的轉(zhuǎn)錄組學(xué)特征未見(jiàn)報(bào)道。因此,本文以團(tuán)頭魴為研究對(duì)象,經(jīng)過(guò)8周高糖高脂日糧的飼養(yǎng)實(shí)驗(yàn),利用組織切片與轉(zhuǎn)錄組學(xué)的方法來(lái)研究高脂日糧(HFD,脂肪含量12.22%)、高糖日糧(HCBD,還原糖含量34.13%)和高脂高糖日糧(HFHCD,脂肪含量10.43%,還原糖含量30.80%)對(duì)團(tuán)頭魴生長(zhǎng)性能、肝組織及肝轉(zhuǎn)錄組的影響,并在此基礎(chǔ)上,結(jié)合實(shí)時(shí)定量PCR、血液生化指標(biāo)及部分肝... 

【文章來(lái)源】:華中農(nóng)業(yè)大學(xué)湖北省 211工程院校 教育部直屬院校

【文章頁(yè)數(shù)】:154 頁(yè)

【學(xué)位級(jí)別】:博士

【文章目錄】:
中文摘要
ABSTRACT
ABBREVIATIONS
CHAPTER 1 Review of literature
    1.1 General introduction of Megalobrama amblycephala
        1.1.1 Biological characteristics
        1.1.2 Megalobrama amblycephala nutrient requirements
        1.1.3 The problem of Megalobrama amblycephala in productions
    1.2 Transcriptome study
        1.2.1 Introduction of transcriptome
        1.2.2 The ability of transcriptomics
        1.2.3 The use of transcriptome data to explore gene functions
        1.2.4 Transcriptome studies in aquatic animals
        1.2.5 The study of transcriptome in Megalobrama amblycephala
    1.3 Fat and carbohydrate in fish diets
        1.3.1 Fat and carbohydrate characteristics
        1.3.2 Function in animals
        1.3.3 Fat and carbohydrate diets in aquaculture
    1.4 Fatty liver disease
        1.4.1 Fatty liver disease types
        1.4.2 Fatty liver disease in fish researches
    1.5 Objective and significance
        1.5.1 Objective
        1.5.2 Significance
CHAPTER 2 The effect of high fat and high carbohydrate diets on growth and liver histology of Megalobrama amblycephala
    2.1 Introduction
    2.2 Materials and methods
        2.2.1 Animal rearing and experimental procedures
        2.2.2 Dietary preparation and proximate analysis
        2.2.3 Sample collection
        2.2.4 Histology
    2.3 Results
        2.3.1 The effect of high fat and high carbohydrate diets on growth performance
        2.3.2 Liver histology changes of BSB fed with high fat and high carbohydrate diets
    2.4 Discussion
CHAPTER 3 The overview of transcriptome analysis fed with high fat and high carbohydrate diets in Megalobrama amblyphyla
    3.1 Introduction
    3.2 Materials and methods
        3.2.1 Sampling and RNA extraction
        3.2.2 c DNA library preparation and Illumina sequencing
        3.2.3 De novo assembly of sequencing reads
        3.2.4 Annotation
        3.2.5 Gene expression analysis
        3.2.6 SSR markers
    3.3 Results
        3.3.1 Transcriptome sequencing and assembly
        3.3.2 Blast search analysis and unigene functional annotation
        3.3.3 The differentially expressed gene profile
        3.3.4 Pathway analysis
        3.3.5 SSR markers
    3.4 Discussion
CHAPTER 4 High Carbohydrate Diet Negatively Affects the Liver Health of Megalobrama amblycephala
    4.1 Introduction
    4.2 Materials and methods
        4.2.1 Diet preparation and proximate analysis
        4.2.2 Animal rearing, experimental procedures and sample collection
        4.2.3 Metabolomics study by NMR
        4.2.4 Transcriptome analysis
        4.2.5 qRT-PCR analysis
    4.3 Results
        4.3.1 Hepatosomatic index and liver histology
        4.3.2 Serum biochemistry
        4.3.3 The metabolomics of serum and liver
        4.3.4 Impacts of high-carbohydrate diet on the expressions of DEGs associated with NAFLD and insulin signaling pathways
    4.4 Discussions
        4.4.1 High-carbohydrate induced hepatosomatic index and liver histology changes
        4.4.2 High-carbohydrate induced changes in serum biochemistry
        4.4.3 High-carbohydrate induced fatty liver associated changes in serum metabolomics and liver metabolomics
        4.4.4 The expression of DEGs in NAFLD and insulin signaling pathways via transcriptome and qRT-PCR
CHAPTER 5 High-fat-high-carbohydrate diet causes mitochondrial dysfunction in the liver of Megalobrama amblycephala
    5.1 Introduction
    5.2 Materials and methods
        5.2.1 Diet preparation and proximate analysis
        5.2.2 Animal rearing, feeding and sample collection
        5.2.3 Transcriptome analysis
            5.2.3.1 RNA extraction
            5.2.3.2 cDNA library preparation and Illumina sequencing
            5.2.3.3 De novo assembly of sequencing reads
            5.2.3.4 Annotation
            5.2.3.5 Ontology analysis
            5.2.3.6 Gene expression analysis
        5.2.4 qRT-PCR analysis
    5.3 Results
        5.3.1 Transcriptome sequencing and annotation of unigenes
        5.3.2 Differential expression analysis
        5.3.3 qRT-PCR
    5.4 Discussion
CONCLUSIONS
REFERENCES
APPENDIX 1
    Fig. S1 Metabolomics graphs
    Fig. S2 ~1H NMR spectra
    Fig. S3 KEGG pathways
    Fig. S4 Alzheimer’s disease
    Fig. S5 Huntington’s disease
    Fig. S6 Parkinson’s disease
    Table S1. ~1H NMR signal assignment of metabolites in serum and liver extracts
    Table S2. Differentially expressed genes
APPENDIX 2 Publications
APPENDIX 3 Awards and Funding
ACKNOWLEDGEMENTS


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