蛇床子素和葛根素对福美双诱导的肉鸡胫骨发育不良的治疗作用及分子机制研究
发布时间：2021-12-10 11:23
胫骨软骨发育不良（TD）是一种影响快速生长期肉鸡的骨骼异常发育疾病。除了对全世界的禽业造成巨大的经济损失外,TD也引起了对动物福利的关注。TD的特点是在胫骨的近端生长板处存在无血管化,无矿化的白色不透明软骨栓。患TD的鸡表现出不良的身体状况,畸形的胫骨以及跛行。1.蛇床子素:一种香豆素衍生物通过调节鸡的BMP-2和RUNX-2表达缓解福美双所致的胫骨软骨发育不良禽胫骨软骨发育不良发生于快速生长的肉鸡,约占肉鸡腿病的30%。本研究旨在评估蛇床子素对禽类胫骨软骨发育不良（TD）的疗效。将240只鸡平均分为对照组,TD组和蛇床子素治疗（osthole）组（n=80）。TD和osthole组的鸡在4–7天以添加50mg/kg饲料重的四甲基秋兰姆二硫（thiram）的饲料进行饲喂,然后在8–18天通过口服的方式给予鸡20 mg/kg体重的蛇床子素。饲喂Thiram会导致跛行,增加死亡率,并降低生产参数,碱性磷酸酶（ALP）,超氧化物歧化酶（SOD）,总抗氧化能力（TAOC）,谷胱甘肽过氧化物酶（GSH-Px）,天冬氨酸氨基转移酶（AST）,丙氨酸氨基转移酶（ALT）,丙二醛（MDA）的水平和并使...
【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】：87 页
【学位级别】：博士
【文章目录】：
摘要
ABSTRACT
CHAPTER 1 General Introduction
1.1.Endochondral bone formation and ossification
1.2.Epiphyseal growth plate
1.2.1.Reserve zone
1.2.2.Proliferative zone
1.2.3.Transition zone
1.2.4.Prehypertrophic zone
1.2.5.Hypertrophic zone
1.3.Avian growth plate
1.4.Tibial dyschondroplasia(TD)
1.5.Tetramethylthiuram disulfide(Thiram)
1.6.TD and gene expression
1.6.1.Role of Bone morphogenetic protein-2(BMP-2)in TD
1.6.2.Role of Runt-related transcription factor-2(RUNX-2)in TD
1.6.3.Role of Hypoxia Inducible Factor-1α (HIF-1α)in TD
1.6.4.Role of Tissue inhibitor of metalloproteinase-3(TIMP-3)in TD
1.6.5.Role of B-cell lymphoma-2(BCL-2)in TD
1.7.Prevention and Treatment of TD
1.8.Traditional Chinese Medicines
1.8.1.Osthole
1.8.2.Puerarin
1.9.Conclusion and Hypothesis
CHAPTER 2 Osthole:A Coumarin Derivative Assuage Thiram-Induced Tibial Dyschondroplasia by Regulating BMP-2 and RUNX-2 Expressions in Chickens
2.1.Introduction
2.2.Objectives
2.3.Materials and methods
2.3.1.Animal ethics
2.3.2.Experimental Birds,Chemical Reagents,and Medicine
2.3.3.Experiment Design
2.3.4.Samples Collection
2.3.5.Mortality Assay,Production Parameters,and Visceral Organs Indices
2.3.6.Biochemical and Antioxidants Analyses
2.3.7.Tibia Bone Parameters
2.3.8.Hematoxylin and Eosin(H&E)Staining and Immunohistochemistry
2.3.9.Reverse Transcription Quantitative Polymerase Chain Reaction(RT-q PCR)
2.3.10.Western Blotting
2.3.11.Statistical Analyses
2.4.Results
2.4.1.Clinical Observations of Thiram–Induced TD
2.4.2.Chicken Mortality and Survival Rate
2.4.3.Production Parameters Analysis
2.4.4.Tibia Parameters Analysis
2.4.5.Serum Biochemical and Liver Antioxidants Analyses
2.4.6.Visceral Organs Indices
2.4.7.Histological Examination of the Tibial Growth Plates
2.4.8.Immunohistochemical Analysis
2.4.9.The m RNA and Protein Expressions of BMP-2 and RUNX-2
2.5.Discussion
CHAPTER 3 Puerarin enhance vascular proliferation and halt apoptosis in thiram-induced avian tibial dyschondroplasia by regulating HIF-1α,TIMP-3 and BCL-2 expressions
3.1.Introduction
3.2.Objectives
3.3.Material and methods
3.3.1.Ethical approval,TD induction and puerarin administration
3.3.2.Samples collection
3.3.3.Percent survival,production and visceral organ’parameters
3.3.4.Serum biochemical and antioxidant analyses
3.3.5.Tibia bone morphometry
3.3.6.Hematoxylin& Eosin(H&E)staining and immunohistochemistry
3.3.7.Reverse transcription quantitative polymerase chain reaction(RT-q PCR)
3.3.8.Western blot
3.3.9.Statistical analyses
3.4.Results
3.4.1.Clinical signs,percent survival and mortality
3.4.2.Puerarin enhance chicken production parameters
3.4.3.Puerarin assuage hepatic dysfunction and oxidative stress
3.4.4.Puerarin impact on visceral organ weight and indices
3.4.5.Tibia bone morphometry and growth plate morphology
3.4.6.Histopathology of the tibial growth plate
3.4.7.Immunohistochemical analysis
3.4.8.Expression of HIF-1α,TIMP-3 and BCL-2
3.5.Discussion
CHAPTER 4 Conclusion and future prospects
4.1.Study I
4.2.Study II
4.3.Future prospects
REFERENCES
ACKNOWLEDGEMENT
SCIENTIFIC CONTRIBUTION
本文编号：3532526
【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】：87 页
【学位级别】：博士
【文章目录】：
摘要
ABSTRACT
CHAPTER 1 General Introduction
1.1.Endochondral bone formation and ossification
1.2.Epiphyseal growth plate
1.2.1.Reserve zone
1.2.2.Proliferative zone
1.2.3.Transition zone
1.2.4.Prehypertrophic zone
1.2.5.Hypertrophic zone
1.3.Avian growth plate
1.4.Tibial dyschondroplasia(TD)
1.5.Tetramethylthiuram disulfide(Thiram)
1.6.TD and gene expression
1.6.1.Role of Bone morphogenetic protein-2(BMP-2)in TD
1.6.2.Role of Runt-related transcription factor-2(RUNX-2)in TD
1.6.3.Role of Hypoxia Inducible Factor-1α (HIF-1α)in TD
1.6.4.Role of Tissue inhibitor of metalloproteinase-3(TIMP-3)in TD
1.6.5.Role of B-cell lymphoma-2(BCL-2)in TD
1.7.Prevention and Treatment of TD
1.8.Traditional Chinese Medicines
1.8.1.Osthole
1.8.2.Puerarin
1.9.Conclusion and Hypothesis
CHAPTER 2 Osthole:A Coumarin Derivative Assuage Thiram-Induced Tibial Dyschondroplasia by Regulating BMP-2 and RUNX-2 Expressions in Chickens
2.1.Introduction
2.2.Objectives
2.3.Materials and methods
2.3.1.Animal ethics
2.3.2.Experimental Birds,Chemical Reagents,and Medicine
2.3.3.Experiment Design
2.3.4.Samples Collection
2.3.5.Mortality Assay,Production Parameters,and Visceral Organs Indices
2.3.6.Biochemical and Antioxidants Analyses
2.3.7.Tibia Bone Parameters
2.3.8.Hematoxylin and Eosin(H&E)Staining and Immunohistochemistry
2.3.9.Reverse Transcription Quantitative Polymerase Chain Reaction(RT-q PCR)
2.3.10.Western Blotting
2.3.11.Statistical Analyses
2.4.Results
2.4.1.Clinical Observations of Thiram–Induced TD
2.4.2.Chicken Mortality and Survival Rate
2.4.3.Production Parameters Analysis
2.4.4.Tibia Parameters Analysis
2.4.5.Serum Biochemical and Liver Antioxidants Analyses
2.4.6.Visceral Organs Indices
2.4.7.Histological Examination of the Tibial Growth Plates
2.4.8.Immunohistochemical Analysis
2.4.9.The m RNA and Protein Expressions of BMP-2 and RUNX-2
2.5.Discussion
CHAPTER 3 Puerarin enhance vascular proliferation and halt apoptosis in thiram-induced avian tibial dyschondroplasia by regulating HIF-1α,TIMP-3 and BCL-2 expressions
3.1.Introduction
3.2.Objectives
3.3.Material and methods
3.3.1.Ethical approval,TD induction and puerarin administration
3.3.2.Samples collection
3.3.3.Percent survival,production and visceral organ’parameters
3.3.4.Serum biochemical and antioxidant analyses
3.3.5.Tibia bone morphometry
3.3.6.Hematoxylin& Eosin(H&E)staining and immunohistochemistry
3.3.7.Reverse transcription quantitative polymerase chain reaction(RT-q PCR)
3.3.8.Western blot
3.3.9.Statistical analyses
3.4.Results
3.4.1.Clinical signs,percent survival and mortality
3.4.2.Puerarin enhance chicken production parameters
3.4.3.Puerarin assuage hepatic dysfunction and oxidative stress
3.4.4.Puerarin impact on visceral organ weight and indices
3.4.5.Tibia bone morphometry and growth plate morphology
3.4.6.Histopathology of the tibial growth plate
3.4.7.Immunohistochemical analysis
3.4.8.Expression of HIF-1α,TIMP-3 and BCL-2
3.5.Discussion
CHAPTER 4 Conclusion and future prospects
4.1.Study I
4.2.Study II
4.3.Future prospects
REFERENCES
ACKNOWLEDGEMENT
SCIENTIFIC CONTRIBUTION
本文编号：3532526
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