基于PI3K/AKT/HIF-1α信号通路研究传统中药治疗肉鸡胫骨软骨发育不良的作用机制
发布时间：2022-01-17 09:23
胫骨软骨发育不良（Tibial dyschondroplasia,TD）是家禽较为常见的骨骼疾病,主要发生于快大型、速生长的肉鸡、火鸡等。TD主要病理学特征是病鸡胫骨近干骺端干骺面生长板内出现无血管化、无钙化的白色“软骨栓”。该病主要引起腿部疾患,在商品肉鸡中最为常见,严重降低肉的品质,造成巨大的经济损失,目前该病已成为家禽跛足的主要原因,严重影响动物福利。1.基于HIF-1α/VEGF信号通路研究川芎嗪对胫骨软骨发育不良的影响肉鸡胫骨软骨发育不良（TD）是许多禽类常见的骨骼疾病,其特征是生长板（GP）构建异常,导致GP内持续无血管、无钙化病变。川芎嗪（TMP）被广泛应用于神经血管疾病和肺动脉高压的治疗,但TMP对肉鸡TD的是否具有一定的治疗作用,尚未见报道。因此,本研究随机选择AA肉鸡210只,饲喂标准日粮;于第三天随机将其分为3组:对照组（n=70）给予标准日粮,福美双组（n=70）和TMP组（n=70）从第4天至第7天,均以饲料添加福美双（thiram,50mg/kg）诱导TD。第8天,TD组与对照组一样给予标准日粮,TMP组给予TMP（30mg/kg/d）,直至试验结束,分别于...
【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】：119 页
【学位级别】：博士
【文章目录】：
摘要
ABSTRACT
LIST OF ABBREVIATIONS
CHAPTER 1 General Introduction
1.1 Endochondral bone formation and ossification
1.2 Long bone growth and development
1.3 Epiphyseal growth plate
1.3.1 Reserve zone of GP
1.3.2 Proliferative zone of GP
1.3.3 Transition zone of GP
1.3.4 Prehypertrophic zone of GP
1.3.5 Hypertrophic zone of GP
1.4 Avian growth plate
1.5 Tibial dyschondroplasia
1.6 Tetramethylthiuram disulfide (Thiram)
1.7 TD and gene expression
1.8 Role of hypoxia inducible factor-1α in TD
1.9 Role of VEGF and VEGFR1 in TD
1.10 Role of PI3K and Akt
1.11 Role of Cyclooxygenase-2
1.12 Role of integrin beta 3
1.13 Prevention and treatment of TD
1.14 Traditional Chinese medicines
1.14.1 Tetramethylpyrazine
1.14.2 Astragaloside IV
1.15 Conclusion and hypothesis
CHAPTER 2 Effects of tetramethylpyrazine on tibial dyschondroplasia by regulating HIF-1α and VEGF
2.1 Introduction
2.2 Objectives
2.3 Materials and Methods
2.3.1 Animal ethics and welfare
2.3.2 Chicken management and experimental design
2.3.3 Morphological, production parameters analysis and sample collection
2.3.4 Hematoxylin and eosin (H&E) staining
2.3.5 Immunohistochemistry
2.3.6 Quantitative-real time PCR (RT-qPCR)
2.3.7 Western blot analysis
2.3.8 Statistical analysis
2.4 Results
2.4.1 TMP prevented the mortality in chickens
2.4.2 Performance parameters analysis of chickens
2.4.3 Effect of TMP on TD score
2.4.4 The overall tibial parameters analysis
2.4.5 Clinical observation of thiram-induced TD and TMP chickens
2.4.6 Histological examination of the tibial growth plates
2.4.7 Immunohistochemistry of tibial growth plates
2.4.8 The expression of HIF-1α/VEGF genes and proteins level in growth plate
2.4.9 Overall effect of TMP on TD chickens
2.5 Discussion
CHAPTER 3 Ligustrazine remodels the toxicity in thiram-induced tibial dyschondroplasia chickens: involvement of new molecules modulating integrin beta-3
3.1 Introduction
3.2 Objectives
3.3 Materials and Methods
3.3.1 Animal ethics
3.3.2 Experimental design and treatment
3.3.3 Incidence and severity of TD
3.3.4 Serum analysis
3.3.5 Measurement of liver antioxidant enzymes
3.3.6 Hematoxylin & Eosin (H&E) and immunofluorescence staining
3.3.7 Tibia index
3.3.8 Quantitative real time polymerase chain reaction (RT-qPCR)
3.3.9 Western blot analysis
3.3.10 Statistical analysis
3.4 Results
3.4.1 Chickens performance
3.4.2 Biochemical criterion analysis of serum
3.4.3 Determination of oxidative stress in liver
3.4.4 Tibia bone observation and H&E examination
3.4.5 Incidence and severity of TD
3.4.6 Tibial index
3.4.7 Histopathology micrograph
3.4.8 Expression of ITGB3
3.5 Discussion
CHAPTER 4 Protective effect of astragaloside IV to inhibit tibial dyschondroplasia on vascular endothelial cell based PI3K/Akt/HIF-1α signaling
4.1 Introduction
4.2 Objectives
4.3 Materials and Methods
4.3.1 Ethics approval
4.3.2 Chicken management and experimental design
4.3.3 Production parameter analysis and sample collection
4.3.4 Biochemical criterion determination
4.3.5 Measurement of liver antioxidant enzymes
4.3.6 Hematoxylin and Eosin (H&E) staining
4.3.7 RT-qPCR assay
4.3.8 Western blotting
4.3.9 Immunohistochemistry assay
4.3.10 Statistical analysis
4.4 Results
4.4.1 Production parameter analysis
4.4.2 H&E staining
4.4.3 Tibial parameters analysis
4.4.4 Biochemical criterion determination
4.4.5 Antioxidant ability of AST-IV
4.4.6 Effect of AST-IV on overall performance of chickens
4.4.7 RT-qPCR assay
4.4.8 Western blotting analysis
4.4.9 Immunohistochemistry analysis
4.4.10 Astragaloside IV regulates PI3K/Akt/HIF-1α signaling pathway
4.5 Discussion
CHAPTER 5 Conclusions and Future prospectus
5.1 Conclusion
5.2 Future prospectus
REFERENCES
Acknowledgement
Publications
【参考文献】：
期刊论文
[1]黄芪皂苷对化疗贫血小鼠PI3K/Akt/mTOR信号通路相关基因mRNA表达的影响[J]. 乔铁,梁可,马进,林蔗茹,郑冰元,张立德. 中国中药杂志. 2016(20)
[2]The role of vascular endothelial growth factor in ossification[J]. Ricky Wong,Alex Wenden,A Bakr M Rabie. International Journal of Oral Science. 2012(02)
本文编号：3594478
【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】：119 页
【学位级别】：博士
【文章目录】：
摘要
ABSTRACT
LIST OF ABBREVIATIONS
CHAPTER 1 General Introduction
1.1 Endochondral bone formation and ossification
1.2 Long bone growth and development
1.3 Epiphyseal growth plate
1.3.1 Reserve zone of GP
1.3.2 Proliferative zone of GP
1.3.3 Transition zone of GP
1.3.4 Prehypertrophic zone of GP
1.3.5 Hypertrophic zone of GP
1.4 Avian growth plate
1.5 Tibial dyschondroplasia
1.6 Tetramethylthiuram disulfide (Thiram)
1.7 TD and gene expression
1.8 Role of hypoxia inducible factor-1α in TD
1.9 Role of VEGF and VEGFR1 in TD
1.10 Role of PI3K and Akt
1.11 Role of Cyclooxygenase-2
1.12 Role of integrin beta 3
1.13 Prevention and treatment of TD
1.14 Traditional Chinese medicines
1.14.1 Tetramethylpyrazine
1.14.2 Astragaloside IV
1.15 Conclusion and hypothesis
CHAPTER 2 Effects of tetramethylpyrazine on tibial dyschondroplasia by regulating HIF-1α and VEGF
2.1 Introduction
2.2 Objectives
2.3 Materials and Methods
2.3.1 Animal ethics and welfare
2.3.2 Chicken management and experimental design
2.3.3 Morphological, production parameters analysis and sample collection
2.3.4 Hematoxylin and eosin (H&E) staining
2.3.5 Immunohistochemistry
2.3.6 Quantitative-real time PCR (RT-qPCR)
2.3.7 Western blot analysis
2.3.8 Statistical analysis
2.4 Results
2.4.1 TMP prevented the mortality in chickens
2.4.2 Performance parameters analysis of chickens
2.4.3 Effect of TMP on TD score
2.4.4 The overall tibial parameters analysis
2.4.5 Clinical observation of thiram-induced TD and TMP chickens
2.4.6 Histological examination of the tibial growth plates
2.4.7 Immunohistochemistry of tibial growth plates
2.4.8 The expression of HIF-1α/VEGF genes and proteins level in growth plate
2.4.9 Overall effect of TMP on TD chickens
2.5 Discussion
CHAPTER 3 Ligustrazine remodels the toxicity in thiram-induced tibial dyschondroplasia chickens: involvement of new molecules modulating integrin beta-3
3.1 Introduction
3.2 Objectives
3.3 Materials and Methods
3.3.1 Animal ethics
3.3.2 Experimental design and treatment
3.3.3 Incidence and severity of TD
3.3.4 Serum analysis
3.3.5 Measurement of liver antioxidant enzymes
3.3.6 Hematoxylin & Eosin (H&E) and immunofluorescence staining
3.3.7 Tibia index
3.3.8 Quantitative real time polymerase chain reaction (RT-qPCR)
3.3.9 Western blot analysis
3.3.10 Statistical analysis
3.4 Results
3.4.1 Chickens performance
3.4.2 Biochemical criterion analysis of serum
3.4.3 Determination of oxidative stress in liver
3.4.4 Tibia bone observation and H&E examination
3.4.5 Incidence and severity of TD
3.4.6 Tibial index
3.4.7 Histopathology micrograph
3.4.8 Expression of ITGB3
3.5 Discussion
CHAPTER 4 Protective effect of astragaloside IV to inhibit tibial dyschondroplasia on vascular endothelial cell based PI3K/Akt/HIF-1α signaling
4.1 Introduction
4.2 Objectives
4.3 Materials and Methods
4.3.1 Ethics approval
4.3.2 Chicken management and experimental design
4.3.3 Production parameter analysis and sample collection
4.3.4 Biochemical criterion determination
4.3.5 Measurement of liver antioxidant enzymes
4.3.6 Hematoxylin and Eosin (H&E) staining
4.3.7 RT-qPCR assay
4.3.8 Western blotting
4.3.9 Immunohistochemistry assay
4.3.10 Statistical analysis
4.4 Results
4.4.1 Production parameter analysis
4.4.2 H&E staining
4.4.3 Tibial parameters analysis
4.4.4 Biochemical criterion determination
4.4.5 Antioxidant ability of AST-IV
4.4.6 Effect of AST-IV on overall performance of chickens
4.4.7 RT-qPCR assay
4.4.8 Western blotting analysis
4.4.9 Immunohistochemistry analysis
4.4.10 Astragaloside IV regulates PI3K/Akt/HIF-1α signaling pathway
4.5 Discussion
CHAPTER 5 Conclusions and Future prospectus
5.1 Conclusion
5.2 Future prospectus
REFERENCES
Acknowledgement
Publications
【参考文献】：
期刊论文
[1]黄芪皂苷对化疗贫血小鼠PI3K/Akt/mTOR信号通路相关基因mRNA表达的影响[J]. 乔铁,梁可,马进,林蔗茹,郑冰元,张立德. 中国中药杂志. 2016(20)
[2]The role of vascular endothelial growth factor in ossification[J]. Ricky Wong,Alex Wenden,A Bakr M Rabie. International Journal of Oral Science. 2012(02)
本文编号：3594478
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