百里香醌及白桦酯醇通过激活LKB1-AMPK信号通路抑制肝纤维化以及酒精性脂肪肝的机制研究

发布时间：2021-01-11 21:00

　　目的：肝纤维化是继发于各种慢性致病因素引起的肝脏炎症损伤后组织修复过程中的代偿反应,以细胞外基质的过度沉积为病理特征,是各种慢性肝病向肝硬化发展所共有的病理改变和必经途径。而肝纤维化发生的中心环节就是肝星状细胞的激活。在肝损伤的过程中,活化的肝星状细胞是肝纤维化形成过程中产生胶原的主要细胞。此外,长期过量饮酒可导致酒精性肝病的发生,使细胞外基质和胶原蛋白过度沉积,进而发展成肝纤维化以及肝硬化。酒精性肝病是世界范围内慢性肝病的最重要病因之一,严重危害人民健康。严重酗酒时可诱发广泛肝细胞坏死或肝功能衰竭,甚至会加剧慢性病毒性肝炎以及非酒精性肝病的发生。因此,发现更多新源药物更深入的研究抗肝纤维化作用靶点是肝纤维化治疗的迫切需要。本研究采用硫代乙酰胺诱导慢性肝纤维化模型和慢性酒精喂养加急性酒精灌胃的酒精性肝病小鼠模型,通过体外、体内实验研究百里香醌以及白桦酯醇的肝保护作用及潜在的作用机制。方法：（1）百里香醌通过调节TLR4及LKB1-AMPK信号通路抑制肝纤维化实验研究。体外实验中用不同浓度的百里香醌预处理肝星状细胞（T-HSC/CI-6）1小时后,加入1 pg/ml内毒素（LPS）孵育2... 

【文章来源】：延边大学吉林省 211工程院校

【文章页数】：109 页

【学位级别】：博士

【文章目录】：
List of Figures
Abbreviations
Abstract in Chinese
Abstract
Introduct
    1. Chronic hepatic injury
    2. Hepatic stellate cells in hepatic fibrosis
    3. TLR4 signaling pathways in liver fibrosis
    4. LKB1-AMPK signaling pathways in hepatic injury
    5. Traditional Chinese medicine in liver disease
Chapter 1.Thymoquinone attenuates liver fibrosis via TLR4 and LKB1-AMPKsignaling pathways
    Abstract
    1.1 Introduction
    1.2 Materials and Methods
        1.2.1 Materials
        1.2.2 In vitro study
            1.2.2.1 Cell cultures
            1.2.2.2 Measurement of cell viability by MTT assay
        1.2.3 In vivo study
            1.2.3.1 Animals and treatments
            1.2.3.2 Serum biochemical parameters assay
            1.2.3.3 Histopathological analysis
            1.2.3.4 Immunohistochemistry examination
            1.2.3.5 Reverse Transcription Polymerase Chain Reaction（RT-PCR）
        1.2.4 Western blot analysis
        1.2.5 Statistical analysis
    1.3 Results
        1.3.1 In vitro results
            1.3.1.1 Effects of TQ on the Viability
            1.3.1.2 Effects of TQ on protein expression of CD14 and TLR4
            1.3.1.3 Effects of TQ on HSCs apoptosis
            1.3.1.4 Effects of TQ on protein expression of collagen-I and α-SMA
            1.3.1.5 Effects of TQ on LPS-induced protein expression of PI3K/Akt phosphorylation
        1.3.2 In vivo results
            1.3.2.1 Effects of TQ on ALT and AST activities
            1.3.2.2 Histopathological and immunohistochemical changes in mice livers after TAA treatment
            1.3.2.3 Effects of TQ on the expression of collagen-I,α-SMA and TIMP-1
            1.3.2.4 Effects of TQ on the expression of TLR4
            1.3.2.5 Effects of TQ on TAA treated protein expression of PI3K phosphorylation
            1.3.2.6 Effects of TQ on the activation of AMPK signaling pathways in TAA induced liver fibrosis
            1.3.2.7 Effects of TQ on proinflammatory cytokines levels
    1.4 Discussion
Chapter 2.Betulin alleviated ethanol-induced alcoholic liver steatosis throughLKB1-AMPK signaling pathways
    Abstract
    2.1 Introduction
    2.2 Materials and Methods
        2.2.1 Materials
        2.2.2 In vitro study
            2.2.2.1 Cell cultures
            2.2.2.2 Measurement of cell viability by MTT assay
        2.2.3 In vivo study
            2.2.3.1 Animals
            2.2.3.2 Chronic-binge ethanol feeding model
            2.2.3.3 Serum biochemical parameters and triglyceride assay
            2.2.3.4 Histopathological analysis
            2.2.3.5 Immunohistochemistry examination
            2.2.3.6 Immunofluorescence imaging assay
        2.2.4 Western blot analysis
        2.2.5 Statistical analysis
    2.3
        2.3.1
            2.3.1.3 AMPK signaling is implicated in the supp ression of ethanol-induced SREBP-1 expression by BT in LX-2 cells
            2.3.1.4 LKB1 signaling is implicated in the suppression of ethanol-induced SREBP-1 expression by BT in LX-2 cells
            2.3.1.5 Effects of BT on ethanol-induced protein expression of LKB1-AMPK phosphorylation
            2.3.1.6 Effect of BT on ethanol-induced SIRT1 expression
            2.3.1.7 Effects of BT on ethanol-induced collagen-I and α-SMA levels in LX-2 cells
        2.3.2 In vivo results
            2.3.2.1 Effects of BT on liver weight induced by chronic-binge ethanol
            2.3.2.2 Effects of BT on serum biochemical parameters and triglyceride assay
            2.3.2.3 Effects of BT on histopathological and Immunohistochemical changes induced by chronic-binge ethanol in mice
            2.3.2.4 Effect of BT on ch ronic-binge ethanol induced SREBP-1 expression
            2.3.2.5 Effects of BT on chronic-binge ethanol induced protein expression of LKB1-AMPK phosphorylation
            2.3.2.6 Effect of BT on chronic-binge ethanol induced SIRT1 expression
    2.4 Discussion
Conclusions
References
Publications
致谢


【参考文献】：
期刊论文
[1]Pathogenesis of alcoholic liver disease:Role of oxidative metabolism[J]. Elisabetta Ceni,Tommaso Mello,Andrea Galli.  World Journal of Gastroenterology. 2014(47)
[2]Sophocarpine attenuates liver fibrosis by inhibiting the TLR4 signaling pathway in rats[J]. Hui Qian,Jian Shi,Ting-Ting Fan,Jiao Lv,Si-Wen Chen,Chun-Yan Song,Zhi-Wu Zheng,Wei-Fen Xie,Yue-Xiang Chen.  World Journal of Gastroenterology. 2014(07)
[3]Role of ethanol in the regulation of hepatic stellate cell function[J]. Robert G Batey,Jacob George.  World Journal of Gastroenterology. 2006(43)



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