摘要
目的分析共培养时大鼠血管内皮细胞(VECs)对大鼠血管平滑肌细胞(VSMCs)表型转化的影响。方法 VSMCs直接种植于培养板表面,VECs则种植于与VSMCs相对的浮胶底面。免疫荧光方法观察和鉴定VECs和VSMCs,RT-PCR和共聚焦显微镜分析表型相关基因的表达。结果原代培养的大鼠VECs呈铺路石样形态,vWF染色阳性。原代培养的大鼠VSMCs免疫荧光染色α-SMA呈阳性。RT-PCR检测结果表明,48h共培养组VSMCs的合成表型相关基因CRBP-1、Smemb的表达水平显著高于单独培养组,分别为1.4倍、1.5倍;72h达到峰值,分别为1.7倍、2.1倍,96h开始下降;共培养组中收缩表型标记物Smoothelin-B和SM-MHC的基因表达水平在48h、72h显著低于单独培养组,96hSmoothelin-B却高于单独培养组。单独培养组上述各基因的变化趋势不变或保持稳定。免疫荧光结果显示SM-MHC蛋白表达在共培养组中96h后从下降转为升高(P<0.05)。结论在共培养体系中,血管内皮细胞对血管平滑肌细胞表型转化的作用表现为先促进向合成型转化,96h后促进向收缩型转化。
Objective To observe the influence of vascular endothelial cells(VECs) on phenotype alternation of vascular smooth muscle cells (VSMCs). Methods VECs and VSMCs were isolated from thoracic aortas of rat using collagenase digestion and tissue section culturing respectively. A single layer of VSMCs were cultured on the downside of floating gel of Type I collagen from rat tails and faced a single layer of VECs attaching on the surface of cell culture plate. The crosstalk between VECs and VSMCs was allowed throu...
出处
《中国分子心脏病学杂志》
CAS
2011年第2期78-82,共5页
Molecular Cardiology of China
基金
863课题(2009AA02Z110)
关键词
表型
细胞共培养
血管内皮细胞
血管平滑肌细胞
基因表达
Phenotype
Co-culture
vascular smooth muscle cells
vascular endothelial cells
gene expression
