常春藤苷C通过抑制TLR2TLR4及其下游NF—κB和MAPKS的信号级联效应发挥抗炎作用
发布时间：2021-06-25 11:39
在当今社会,随着奶牛对代谢性、感染性和炎性疾病易感性的增加,奶牛养殖业承受着巨大的压力。乳腺炎仍然是奶牛在泌乳期最常见的炎性疾病,导致乳汁产量减少、品质下降、治疗费用增加。据统计,乳腺炎每年造成的经济损失在美国约为18亿美元,在中国约为150—450亿人民币。金黄色葡萄球菌（S.aureus）是最常见和最具感染性的致病菌,可导致危及人类和动物生命的并发症。反刍动物是致病性金黄色葡萄球菌最重要的宿主之一,奶牛是重要的反刍动物同时也是人类重要的食用动物。抗生素是临床上治疗金黄色葡萄球菌性乳腺炎的主要药物,但在治疗过程中产生的抗生素残留及耐药菌又会引发严重的食品安全和公共卫生问题,现在许多国家已被禁止使用。因此,兽医临床实际迫切需要科学有效的防治乳腺炎的药物。中药（Tradition Chinese Medicines TCM）对于细菌性炎性疾病具有良好的治疗效果,在食用动物行业有着广阔的应用前景。本研究目的是阐明常春藤苷-C抑制TLRs-mediated介导的潜在奶牛乳腺炎炎性反应的调节通路及其作用机制,为临床治疗奶牛乳腺炎提供科学依据。1.奶牛乳腺炎临床病理及炎症因子分析乳腺炎是指乳腺组...
【文章来源】：华中农业大学湖北省 211工程院校 教育部直属院校
【文章页数】：163 页
【学位级别】：博士
【文章目录】：
摘要
Abstract
Abbreviations
Chapter 1 Introduction
1.1 Mastitis in Dairy Cows
1.1.1 General Aspects of Bovine Mastitis
1.1.2 Classification of Mastitis
1.1.3 Economic Implications of Mastitis
1.1.4 Epidemiological Percept of Mastitis
1.1.5 Factors Involved in Occurrence and Pathogenesis of Mastitis
1.1.5.1 Host Factors
1.1.5.2 Environmental Factors
1.1.5.3 Pathogens
1.1.6 Clinical Signs and Symptoms of Mastitis
1.1.7 Diagnosis of Mastitis
1.1.8 Strategies for Treatment of Mastitis
1.1.8.1 Genetic Selection of Mastitis Resistance Cows
1.1.8.2 Vaccination
1.1.8.3 Bacteriophage Therapy
1.1.8.4 Lactational Therapy
1.1.8.5 Dry Cow Therapy
1.1.8.6 Use of Teat Sealer
1.1.8.7 Synthetic Drugs (Antibiotics) Should Be Used Carefully
1.1.8.8 Natural Drugs Therapy
1.2 Hederacoside-C to Treat Infectious Diseases
1.2.1 Traditional Chinese Medicines (TCM) to Treat Infections
1.2.2 Effective Ingredients and Therapeutic Effects of Hedera helix Compounds
1.2.3 Hederacoside-C (HDC) and Its Anti-Inflammatory Effects and Mechanisms
1.3 Immune Response in Mastitis
1.3.1 Innate and Acquired Immunity
1.3.2 Mastitis and Immunity Are Interlinked
1.3.3 Defensive Mechanisms Due to Anatomical Structure
1.3.4 Defensive Mechanisms at Cellular Levels
1.3.4.1 Role of Neutrophils
1.3.4.2 Role of Macrophages
1.3.4.3 Role of Lymphocytes
1.3.5 Endogenous Soluble Defensive Mechanism
1.4 Inflammatory Responsive Mediators and Mechanisms in Mastitis
1.4.1 Myeloperoxidase (MPO),A Novel Biomarker for Early Diagnosis
1.4.2 Pattern Recognition Receptors (PRRs)
1.4.2.1 Toll-Like Receptor 2 (TLR2)
1.4.2.2 Toll-Like Receptor 4 (TLR4)
1.4.3 Mitogen-Activated Protein Kinases (MAPKs)
1.4.3.1 p38 MAPK
1.4.3.2 Extracellular Signal Regulated Kinases (ERKs)
1.4.3.3 The c-Jun Amino-terminal Kinases(JNKs)
1.4.4 Nuclear Factor-Kappa B (NF-κB)
1.4.4.1 NF-κB p-65
1.4.4.2 Inhibitor of nuclear factor Kappa-B subunit alpha (IκBα)
1.4.5 Cytokines,the Immune Modulators that Link Cellular Physiology withPathology
1.4.5.1 Pro-inflammatory Cytokines
1.4.5.2 Anti-inflammatory Cytokine
1.5 Objectives and Significance
1.5.1 Statement of Problem
1.5.2 Aims
1.5.3 Project Benefits and Outcomes
Chapter 2 Clinical Histopathology and Inflammatory Factors in Mastitis of DairyCows
2.1 Objectives
2.2 Methodology
2.2.1 Chemicals and Reagents
2.2.2 Target Animals (Cows) and Mammary Gland Tissue Sample Collection
2.2.3 Histopathology
2.2.4 Experimental Groups
2.2.5 Isolation and Identification of S. aureus from Mammary Gland Tissues
2.2.6 Western Blot Analysis for TLRs,NF-κB and MAPKs Proteins
2.2.7 Real-Time Quantitative Polymerase Chain Reaction for Pro-InflammatoryCytokines
2.2.8 Statistical Analysis
2.3 Results
2.3.1 Histopathological Evaluation of Mammary Glands
2.3.2 S. aureus Characterization and Identification
2.3.3 Expressions of TLR2 and TLR4 in Mammary Glands
2.3.4 Phosphorylation of p-38,ERK and JNK in Mammary Glands
2.3.5 Phosphorylation of p-65 and IκBα in Mammary Glands
2.3.6 mRNA Expression Levels of TNF-α, IL-1β and IL-6 in Mammary Glands
2.4 Discussion
Chapter 3 Hederacoside-C Inhibiting Inflammatory Response via TLR2 & TLR4and Their Downstream Signalling NF-κB & MAPKs Pathways in-vitro
3.1 Objectives
3.2 Materials and Methods
3.2.1 Chemicals and Reagents
3.2.2 The Multiplication and Growth of Staphylococcus aureus
3.2.3 High Performance Liquid Chromatography
3.2.4 Cell Viability Assay
3.2.5 Cell Culturing and Treatment Groups
3.2.6 Enzyme-Linked Immunosorbent Assay (ELISA)
3.2.7 Real-Time Quantitative Polymerase Chain Reaction
3.2.8 Western Blot Analysis
3.2.9 Statistics
3.3 Results
3.3.1 Purity of HDC and S. aureus Growth upon HDC Administration
3.3.2 Effect of HDC on RAW 264.7 Cell Viability
3.3.3 HDC modulates Protein/Gene Expression Levels of Inflammatory Cytokinein S. aureus-induced Inflammatory RAW 264.7 cells
3.3.4 HDC inhibits TLR2 & TLR4 Expressions in S. aureus-inducedInflammation RAW 264.7 cells
3.3.5 HDC attenuates NF-κB cascade in S. aureus-induced Inflammation RAW264.7 cells
3.3.6 HDC attenuates MAPKs cascade in S. aureus-induced Inflammation RAW264.7 cells
3.4 Discussion
Chapter 4 Hederacoside-C Suppressing Inflammatory Response in Mastitis viaInhibiting TLR2 & TLR4 and Their Downstream Signalling NF-κB & MAPKsPathways in-vivo
4.1 Objectives
4.2 Materials and Methods
4.2.1 Chemicals and Reagents
4.2.2 Selection of Experimental Animals and Treatment Groups
4.2.3 Histopathological Evaluation
4.2.4 Measurement of Myeloperoxidase (MPO) in the Mammary Glands
4.2.5 Enzyme-Linked Immunosorbent Assay (ELISA)
4.2.6 Real-Time Quantitative Polymerase Chain Reaction (qRT-PCR)
4.2.7 Analysis by Western Blotting
4.2.8 Statistical Analysis
4.3 Results
4.3.1 Histopathological Changes and Analysis of Mammary Glands
4.3.2 Effects of HDC on the Myeloperoxidase Activity
4.3.3 Effects of HDC on the Inflammatory Cytokines in S. aureus-inducedMastitis
4.3.4 Effects of HDC on TLR2 & TLR4 Expression in S. aureus-induced Mastitis
4.3.5 HDC Suppresses the NF-κB Pathway in S. aureus-induced Mastitis
4.3.6 HDC Suppresses the MAPKs pathway in S. aureus-induced Mastitis
4.4 Discussion
Chapter 5 Discussion and Conclusion
Discussion
Conclusion
Achieved Goals
Future Suggestions
References
List of Figures
List of Tables
Acknowledgement
Appendix Ⅰ Publication List and Related Research
Publications List
Related Research
【参考文献】：
期刊论文
[1]The Immune System, Cytokines, and Biomarkers in Autism Spectrum Disorder[J]. Anne Masi,Nicholas Glozier,Russell Dale,Adam J.Guastella. Neuroscience Bulletin. 2017(02)
本文编号：3249147
【文章来源】：华中农业大学湖北省 211工程院校 教育部直属院校
【文章页数】：163 页
【学位级别】：博士
【文章目录】：
摘要
Abstract
Abbreviations
Chapter 1 Introduction
1.1 Mastitis in Dairy Cows
1.1.1 General Aspects of Bovine Mastitis
1.1.2 Classification of Mastitis
1.1.3 Economic Implications of Mastitis
1.1.4 Epidemiological Percept of Mastitis
1.1.5 Factors Involved in Occurrence and Pathogenesis of Mastitis
1.1.5.1 Host Factors
1.1.5.2 Environmental Factors
1.1.5.3 Pathogens
1.1.6 Clinical Signs and Symptoms of Mastitis
1.1.7 Diagnosis of Mastitis
1.1.8 Strategies for Treatment of Mastitis
1.1.8.1 Genetic Selection of Mastitis Resistance Cows
1.1.8.2 Vaccination
1.1.8.3 Bacteriophage Therapy
1.1.8.4 Lactational Therapy
1.1.8.5 Dry Cow Therapy
1.1.8.6 Use of Teat Sealer
1.1.8.7 Synthetic Drugs (Antibiotics) Should Be Used Carefully
1.1.8.8 Natural Drugs Therapy
1.2 Hederacoside-C to Treat Infectious Diseases
1.2.1 Traditional Chinese Medicines (TCM) to Treat Infections
1.2.2 Effective Ingredients and Therapeutic Effects of Hedera helix Compounds
1.2.3 Hederacoside-C (HDC) and Its Anti-Inflammatory Effects and Mechanisms
1.3 Immune Response in Mastitis
1.3.1 Innate and Acquired Immunity
1.3.2 Mastitis and Immunity Are Interlinked
1.3.3 Defensive Mechanisms Due to Anatomical Structure
1.3.4 Defensive Mechanisms at Cellular Levels
1.3.4.1 Role of Neutrophils
1.3.4.2 Role of Macrophages
1.3.4.3 Role of Lymphocytes
1.3.5 Endogenous Soluble Defensive Mechanism
1.4 Inflammatory Responsive Mediators and Mechanisms in Mastitis
1.4.1 Myeloperoxidase (MPO),A Novel Biomarker for Early Diagnosis
1.4.2 Pattern Recognition Receptors (PRRs)
1.4.2.1 Toll-Like Receptor 2 (TLR2)
1.4.2.2 Toll-Like Receptor 4 (TLR4)
1.4.3 Mitogen-Activated Protein Kinases (MAPKs)
1.4.3.1 p38 MAPK
1.4.3.2 Extracellular Signal Regulated Kinases (ERKs)
1.4.3.3 The c-Jun Amino-terminal Kinases(JNKs)
1.4.4 Nuclear Factor-Kappa B (NF-κB)
1.4.4.1 NF-κB p-65
1.4.4.2 Inhibitor of nuclear factor Kappa-B subunit alpha (IκBα)
1.4.5 Cytokines,the Immune Modulators that Link Cellular Physiology withPathology
1.4.5.1 Pro-inflammatory Cytokines
1.4.5.2 Anti-inflammatory Cytokine
1.5 Objectives and Significance
1.5.1 Statement of Problem
1.5.2 Aims
1.5.3 Project Benefits and Outcomes
Chapter 2 Clinical Histopathology and Inflammatory Factors in Mastitis of DairyCows
2.1 Objectives
2.2 Methodology
2.2.1 Chemicals and Reagents
2.2.2 Target Animals (Cows) and Mammary Gland Tissue Sample Collection
2.2.3 Histopathology
2.2.4 Experimental Groups
2.2.5 Isolation and Identification of S. aureus from Mammary Gland Tissues
2.2.6 Western Blot Analysis for TLRs,NF-κB and MAPKs Proteins
2.2.7 Real-Time Quantitative Polymerase Chain Reaction for Pro-InflammatoryCytokines
2.2.8 Statistical Analysis
2.3 Results
2.3.1 Histopathological Evaluation of Mammary Glands
2.3.2 S. aureus Characterization and Identification
2.3.3 Expressions of TLR2 and TLR4 in Mammary Glands
2.3.4 Phosphorylation of p-38,ERK and JNK in Mammary Glands
2.3.5 Phosphorylation of p-65 and IκBα in Mammary Glands
2.3.6 mRNA Expression Levels of TNF-α, IL-1β and IL-6 in Mammary Glands
2.4 Discussion
Chapter 3 Hederacoside-C Inhibiting Inflammatory Response via TLR2 & TLR4and Their Downstream Signalling NF-κB & MAPKs Pathways in-vitro
3.1 Objectives
3.2 Materials and Methods
3.2.1 Chemicals and Reagents
3.2.2 The Multiplication and Growth of Staphylococcus aureus
3.2.3 High Performance Liquid Chromatography
3.2.4 Cell Viability Assay
3.2.5 Cell Culturing and Treatment Groups
3.2.6 Enzyme-Linked Immunosorbent Assay (ELISA)
3.2.7 Real-Time Quantitative Polymerase Chain Reaction
3.2.8 Western Blot Analysis
3.2.9 Statistics
3.3 Results
3.3.1 Purity of HDC and S. aureus Growth upon HDC Administration
3.3.2 Effect of HDC on RAW 264.7 Cell Viability
3.3.3 HDC modulates Protein/Gene Expression Levels of Inflammatory Cytokinein S. aureus-induced Inflammatory RAW 264.7 cells
3.3.4 HDC inhibits TLR2 & TLR4 Expressions in S. aureus-inducedInflammation RAW 264.7 cells
3.3.5 HDC attenuates NF-κB cascade in S. aureus-induced Inflammation RAW264.7 cells
3.3.6 HDC attenuates MAPKs cascade in S. aureus-induced Inflammation RAW264.7 cells
3.4 Discussion
Chapter 4 Hederacoside-C Suppressing Inflammatory Response in Mastitis viaInhibiting TLR2 & TLR4 and Their Downstream Signalling NF-κB & MAPKsPathways in-vivo
4.1 Objectives
4.2 Materials and Methods
4.2.1 Chemicals and Reagents
4.2.2 Selection of Experimental Animals and Treatment Groups
4.2.3 Histopathological Evaluation
4.2.4 Measurement of Myeloperoxidase (MPO) in the Mammary Glands
4.2.5 Enzyme-Linked Immunosorbent Assay (ELISA)
4.2.6 Real-Time Quantitative Polymerase Chain Reaction (qRT-PCR)
4.2.7 Analysis by Western Blotting
4.2.8 Statistical Analysis
4.3 Results
4.3.1 Histopathological Changes and Analysis of Mammary Glands
4.3.2 Effects of HDC on the Myeloperoxidase Activity
4.3.3 Effects of HDC on the Inflammatory Cytokines in S. aureus-inducedMastitis
4.3.4 Effects of HDC on TLR2 & TLR4 Expression in S. aureus-induced Mastitis
4.3.5 HDC Suppresses the NF-κB Pathway in S. aureus-induced Mastitis
4.3.6 HDC Suppresses the MAPKs pathway in S. aureus-induced Mastitis
4.4 Discussion
Chapter 5 Discussion and Conclusion
Discussion
Conclusion
Achieved Goals
Future Suggestions
References
List of Figures
List of Tables
Acknowledgement
Appendix Ⅰ Publication List and Related Research
Publications List
Related Research
【参考文献】：
期刊论文
[1]The Immune System, Cytokines, and Biomarkers in Autism Spectrum Disorder[J]. Anne Masi,Nicholas Glozier,Russell Dale,Adam J.Guastella. Neuroscience Bulletin. 2017(02)
本文编号：3249147
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