摘要
背景:血管钙化是一种细胞介导的主动的、可调控的复杂生物学过程,血管平滑肌细胞转分化为成骨样细胞发挥着重要作用,其确切机制尚不清楚。目的:探讨尿毒症背景下动脉粥样硬化血管钙化的病理生理机制。方法:采用5/6肾切除法建立尿毒症背景下ApoE-/-小鼠动脉血管钙化模型,苏木精-伊红染色和VonKossa染色观察主动脉组织形态学特点,明确造模成功。应用小鼠全基因组Agilent芯片筛查MAPK信号通路的差异表达基因,实时定量PCR分析验证部分与MAPK信号通路相关的差异表达基因,并结合通路分析来探索MAPK信号通路与血管钙化的内在联系。结果与结论:造模12周后,尿毒症ApoE-/-小鼠主动脉组织形态学特点证实动脉粥样硬化钙化斑块形成。Agilent基因芯片检测结果显示,MAPK信号通路中存在14个差异表达基因,RT-PCR验证结果与芯片检测结果相符合。经KEGG通路分析,ERK1/2信号通路可能在血管钙化的病理生理过程中扮演着重要的角色。说明5/6肾切除ApoE-/-小鼠主动脉钙化斑块形成与MAPK信号通路激活密切相关,该信号通路可能在平滑肌细胞转分化过程中起着至关重要作用。
BACKGROUND： Vascular calcification is recognized as an active and regulated biological process involving osteoblast-like cell transdifferentiation of vascular smooth muscle cells. However, the precise mechanism of vascular calcification is stil unclear. OBJECTIVE： To explore the pathophysiological mechanism of atherosclerotic calcification in uremic mice. METHODS： The animal model of atherosclerotic calcification in Apolipoprotein E knock-out mice was established with 5/6 nephrectomy. Histomorphological changes of aorta sections of mice were evaluted by hematoxylin-eosin staining and Von Kossa staining to confirm atherosclerotic calcification. The differential y expressed genes in mitogen-activated protein kinase pathway were evaluated using mouse whole-genome Agilent chip. Real-time quantitative reverse transcriptase-polymerase chain reaction was used to verify gene expression changes related to mitogen-activated protein kinase pathway, and combined with pathway analysis to explore the relationship between mitogen-activated protein kinase pathway and vascular calcification. RESULTS AND CONCLUSION： The histomorphological changes of aorta sections of uremic Apolipoprotein E knock-out mice indicated atherosclerotic calcification after 12 weeks of modeling. Microarray hybridization identified fourteen differentially expressed genes in the mitogen-activated protein kinase pathway, which have significantly altered their expression levels during atherosclerotic calcification. Reverse transcriptase-polymerase chain reaction results were consistent with the chip validation. The extracel ular signal-regulated kinase 1/2 signal transduction pathway played an important role in vascular calcification, identified by KEGG pathway analysis. Experimental findings indicate that, atherosclerotic calcification in Apolipoprotein E knock-out mice with 5/6 nephrectomy is closely associated with mitogen-activated protein kinase signaling pathway, which plays an important role in smooth muscle phenotypic transition.
出处
《中国组织工程研究》
CAS
CSCD
2013年第20期3618-3625,共8页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金项目(81100157)~~
关键词
组织构建
骨组织构建
动脉粥样硬化
血管钙化
尿毒症
平滑肌细胞
丝裂原活化蛋白激酶
基因芯片
表达谱
MAPK信号通路
形态学变化
差异表达基因
国家自然科学基金
tissue construction
bone tissue construction
atherosclerosis
vascular calcification
uremia
smoothmuscle cells
mitogen activated protein kinase
gene chip
expression profile
mitogen-activated protein kinasesignaling pathway
morphology
differentially expressed gene
National Natural Science Foundation of China
