摘要
为从基因水平研究高血压病大鼠血管重塑机制 ,采用抑制消减杂交法筛选两肾一夹肾血管性高血压大鼠重塑大血管 (胸主动脉 )差异表达基因。经过两轮消减杂交和巢式聚合酶链反应扩增 ,获得了富集的差异表达cDNA片段 ,经Genbank等数据库进行同源比较 ,获得 10余个差异表达的表达序列标签 ;采用Westernblot方法对其中 2个已知基因编码蛋白细胞色素C和Bcl 2的表达进行检测。结果发现细胞色素C在高血压大鼠重塑大血管组织中表达增高 ,而Bcl 2在高血压大鼠重塑大血管组织中表达减少。研究结果提示氧化应激导致细胞凋亡机制尤其是Bcl 2 细胞色素C
Aim Cloning of genes that differentially expressed in remodeling thoracic aorta of 2K1C hypertensive rats(2-kidney, 1-clip Goldblatt rats), which is important for understanding the molecular basis of hypertensive vascular remodeling. Methods We used suppression subtractive hybridization (SSH) to isolate differentially expressed EST in remodeling thoracic aorta. This led to identification of more than fifteen differential expressed sequence-tags(EST). After further cloned and sequenced and homology searched in Genbank with software Stand BLAST. Some differentially displayed genes coding protein were confirmed by western blot. Results We have obtained fifteen pieces of differentially displayed EST, such as cytochrome C and Bcl-2. Moreover, the results from Western Blot confirmed that cytochrome C is highly expressed in in remodeling thoracic aorta and Bcl-2 lowly expressed in remodeling thoracic aorta. Conclusion Cytochrome C is released from mitochondria in response to a variety of apoptotic stimuli, such as reactive oxygen radicals, calcium and ceramide. The Bcl-2 family of proteins serves as critical regulators of pathways involved in apoptosis. Bcl-2 protein is anti-apoptotic. Our data indicate that cell apoptotic mechanism may have important role in hypertensive vascular remodeling.
出处
《中国动脉硬化杂志》
CAS
CSCD
2003年第1期5-8,共4页
Chinese Journal of Arteriosclerosis
基金
国家基础研究重点规划项目 (G2 0 0 0 0 5 690 5 )
湖南省自然科学基金 ( 0 0JJY2 0 2 7)
湖南省教育厅课题 ( 0 2B0 39)资助
关键词
病理学
重塑血管差异表达基因
抑制消减杂交
高血压
胸主动脉
细胞色素C
细胞凋亡
Suppression Subtractive Hybridization
Hypertension
Thoracic Aorta
Cytochrome C
Cell Apoptosis
Vascular Remodeling Related Gene
Gene Expression
