University of Brighton, United Kingdom.
Exp Gerontol. 2009 Oct;44(10):659-65. doi: 10.1016/j.exger.2009.07.004. Epub 2009 Jul 23.
Little is known about the senescent phenotype of human vascular smooth muscle cells (VSMCs) and the potential involvement of senescent VSMCs in age-related vascular disease, such as atherosclerosis. As such, VSMCs were grown and characterised in vitro to generate senescent VSMCs needed for microarray analysis (Affymetrix). Comparative analysis of the transcriptome profiles of early (14 CPD) and late (39-42 CPD) passage VSMCs found a total of 327 probesets called as differentially expressed: 149 are up-regulated in senescence and 178 repressed (p-value<0.5%, minimum effect size of at least 2-fold differential regulation, explore data at http://www.madras.cf.ac.uk/vsmc). Data mining shows a differential regulation of genes at senescence associated with the development of atherosclerosis and vascular calcification. These included genes with roles in inflammation (IL1beta, IL8, ICAM1, TNFAP3, ESM1 and CCL2), tissue remodelling (VEGF, VEGFbeta, ADM and MMP14) and vascular calcification (MGP, BMP2, SPP1, OPG and DCN). The microarray data for IL1beta, IL8 and MGP were validated by either, ELISA, Western blot analysis or RT-PCR. These data thus provide the first evidence for a role of VSMC senescence in the development of vascular calcification and provides further support for the involvement of senescent VSMCs in the progression of atherosclerosis.
目前对于人血管平滑肌细胞（VSMCs）衰老表型知之甚少，衰老 VSMCs 是否参与与年龄相关的血管疾病（如动脉粥样硬化）也尚未可知。因此，本研究通过体外培养 VSMCs 并对其进行特征鉴定，以生成用于微阵列分析（Affymetrix）的衰老 VSMCs。对早期（14 CPD）和晚期（39-42 CPD）传代 VSMCs 的转录组谱进行比较分析，共发现 327 个探针集被称为差异表达：149 个在衰老时上调，178 个下调（p 值<0.5%，最小效应大小至少为 2 倍差异调节，可在 http://www.madras.cf.ac.uk/vsmc 上探索数据）。数据挖掘显示，与动脉粥样硬化和血管钙化发展相关的衰老基因发生差异调节。这些基因包括在炎症（IL1beta、IL8、ICAM1、TNFAP3、ESM1 和 CCL2）、组织重塑（VEGF、VEGFbeta、ADM 和 MMP14）和血管钙化（MGP、BMP2、SPP1、OPG 和 DCN）中起作用的基因。通过 ELISA、Western blot 分析或 RT-PCR 对 IL1beta、IL8 和 MGP 的微阵列数据进行了验证。这些数据首次为 VSMC 衰老在血管钙化发展中的作用提供了证据，并进一步支持衰老 VSMC 参与动脉粥样硬化的进展。
