组织蛋白酶S通过Wnt5a/SC35信号通路调控老龄小鼠缺血诱导的血管再生机制
发布时间：2021-04-16 00:14
目的:本研究探讨老龄小鼠缺血性血管再生过程中CatS介导的Wnt5a/SC35负性调控机制,进一步阐明衰老削弱血管再生的相关分子机制,为防治老年心血管疾病提供实验依据。方法:1、动物实验实验1):老龄小鼠（25-29g,雄性,>24个月龄,n=31）（由日本名古屋大学提供）,分为CatS基因敲除组（CatS-/-,n=16）和野生组（CatS+/+,n=15）。腹腔内注射戊巴比妥（60mg/kg）进行麻醉,脱毛并切开左下腹皮肤,结扎股动脉根部及其主要分支后切除股动脉段的方法制作小鼠下肢缺血模型。造模成功后在特定时间点收集血液,腓肠肌和骨髓样本,开展以下实验:（1）通过激光散斑血流成像（LSBFI）测定术前、术后即刻、4天、7天、14天的下肢血流,用同一时间点的缺血侧与非缺血侧的下肢血流灌注比值分析血流恢复情况;（2）应用CD31和Mac-3抗体进行免疫染色,c-Kit和Ki67抗体进行免疫荧光,评价肌肉组织中的毛细血管形成,巨噬细胞浸润和骨髓中的干细胞增殖。（3）应用流式细胞术（FACS）检测骨髓及外周血中的血管内皮祖细胞（EPC）,评价EPC的增生动员;（4）使用酶联免疫吸附检...
【文章来源】：延边大学吉林省 211工程院校
【文章页数】：60 页
【学位级别】：博士
【文章目录】：
中文摘要
Abstract
Abbreviations （缩略词简表）
Chapter One Introduction
Chapter Two Methods and Materials
2.1 Materials
2.2 Animals
2.3 Mouse hindlimb ischemic model and blood flow analysis
2.4 Gene expression assays
2.5 Immunohistochemical analysis
2.6 Immunoblotting assay
2.7 Gelatin zymography
2.8 Aorta-ring culture assay
2.9 Cell culture
2.10 Immunofluorescence
2.11 Cell migration,invasion,and proliferation assays
2.12 Tubulogenesis assay
2.13 ELISA
2.14 The silencing and the overexpression of CatS
2.15 BM-derived EPC isolation and culture
2.16 Flow cytometry analyses of EPCs
2.17 Statistical analyses
Chapter Three Results
3.1 CatS deficiency impairs angiogenesis in response to ischemia in agedmice
3.2 CatS deletion negatively regulates antiangiogenic Wnt5a/SC35expressions and proangiogenic signaling activation in aged mice andendothelial cells
-/- attenuated microtubule formation ex vivo"> 3.3 CatS-/- attenuated microtubule formation ex vivo
-/- impaired ischemia-induced vasculogenesis"> 3.4 CatS-/- impaired ischemia-induced vasculogenesis
3.5 CatS activity was required for the inflammatory response to ischemicstress in the aged mice
3.6 CatS inhibition suppressed ischemia-induced angiogenesis andvascularization
Chapter Four Discussion
Chapter Five Conclusions
References
致谢
综述
参考文献
附录: 攻读学位期间发表论文目录
本文编号：3140359
【文章来源】：延边大学吉林省 211工程院校
【文章页数】：60 页
【学位级别】：博士
【文章目录】：
中文摘要
Abstract
Abbreviations （缩略词简表）
Chapter One Introduction
Chapter Two Methods and Materials
2.1 Materials
2.2 Animals
2.3 Mouse hindlimb ischemic model and blood flow analysis
2.4 Gene expression assays
2.5 Immunohistochemical analysis
2.6 Immunoblotting assay
2.7 Gelatin zymography
2.8 Aorta-ring culture assay
2.9 Cell culture
2.10 Immunofluorescence
2.11 Cell migration,invasion,and proliferation assays
2.12 Tubulogenesis assay
2.13 ELISA
2.14 The silencing and the overexpression of CatS
2.15 BM-derived EPC isolation and culture
2.16 Flow cytometry analyses of EPCs
2.17 Statistical analyses
Chapter Three Results
3.1 CatS deficiency impairs angiogenesis in response to ischemia in agedmice
3.2 CatS deletion negatively regulates antiangiogenic Wnt5a/SC35expressions and proangiogenic signaling activation in aged mice andendothelial cells
-/- attenuated microtubule formation ex vivo"> 3.3 CatS-/- attenuated microtubule formation ex vivo
-/- impaired ischemia-induced vasculogenesis"> 3.4 CatS-/- impaired ischemia-induced vasculogenesis
3.5 CatS activity was required for the inflammatory response to ischemicstress in the aged mice
3.6 CatS inhibition suppressed ischemia-induced angiogenesis andvascularization
Chapter Four Discussion
Chapter Five Conclusions
References
致谢
综述
参考文献
附录: 攻读学位期间发表论文目录
本文编号：3140359
本文链接：https://www.wllwen.com/yixuelunwen/xxg/3140359.html
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