Weidmann Henri, Touat-Hamici Zahia, Durand Herve, Mueller Christian, Chardonnet Solenne, Pionneau Cedric, Charlotte Frédéric, Janssen Klaus-Peter, Verdugo Ricardo, Cambien Francois, Blankenberg Stefan, Tiret Laurence, Zeller Tanja, Ninio Ewa
Sorbonne Universités, UPMC, UMR_S 1166-ICAN, Genomics and Pathophysiology of Cardiovascular Diseases, Institute of Cardiometabolism and Nutrition, ICAN, Pitié-Salpêtrière Hospital, F-75013, Paris, France; University Heart Center Hamburg, Clinic for General and Interventional Cardiology, Hamburg, Germany; German Center for Cardiovascular Research (DZHK e.V.) Partner Site Hamburg, Lübeck, Kiel, Hamburg, Germany.
Sorbonne Universités, UPMC, UMR_S 1166-ICAN, Genomics and Pathophysiology of Cardiovascular Diseases, Institute of Cardiometabolism and Nutrition, ICAN, Pitié-Salpêtrière Hospital, F-75013, Paris, France.
Atherosclerosis. 2015 Oct;242(2):571-9. doi: 10.1016/j.atherosclerosis.2015.08.013. Epub 2015 Aug 14.
We have previously reported that SASH1 expression is increased in circulating human monocytes from smokers and was positively correlated with the number of carotid atherosclerotic plaques. The aim of this study was to further validate the link between smoking, SASH1 and atherosclerosis within the vascular wall and to assess the impact of SASH1 expression on endothelial cell functions.
Human carotids with atherosclerotic plaques were obtained from 58 patients (45 of them with known smoking status: smoker, non-smoker, ex-smokers), and were processed for gene expression analyses and immunostaining. To investigate its function, SASH1 was silenced in human aortic endothelial cells (HAECs) using two different siRNA and subcellular localization of SASH1 was determined by immunostaining and subcellular fractionation. Subsequently the transcriptomic analyses and functional experiments (wound healing, WST-1 proliferation or Matrigel assays) were performed to characterize SASH1 function.
SASH1 was expressed in human vascular cells (HAECs, smooth muscle cells) and in monocytes/macrophages. Its tissue expression was significantly higher in the atherosclerotic carotids of smokers compared to non-smokers (p < 0.01). In HAECs, SASH1 was expressed mostly in the cytoplasm and SASH1 knockdown resulted in an increased cell migration, proliferation and angiogenesis. Transcriptomic and pathway analyses showed that SASH1 silencing results in a decreased CYP1A1 expression possibly through the inhibition of TP53 activity.
We showed that SASH1 expression is increased in atherosclerotic carotids in smokers and its silencing affects endothelial angiogenic functions; therefore we provide a potential link between smoking and atherosclerosis through SASH1 expression.
我们之前报道过，吸烟者循环血中的人单核细胞中SASH1表达增加，且与颈动脉粥样硬化斑块数量呈正相关。本研究的目的是进一步验证吸烟、SASH1与血管壁内动脉粥样硬化之间的联系，并评估SASH1表达对内皮细胞功能的影响。
从58例患者（其中45例有已知吸烟状态：吸烟者、非吸烟者、既往吸烟者）获取有动脉粥样硬化斑块的人颈动脉，进行基因表达分析和免疫染色。为研究其功能，使用两种不同的小干扰RNA（siRNA）在人主动脉内皮细胞（HAECs）中沉默SASH1，并通过免疫染色和亚细胞分级分离确定SASH1的亚细胞定位。随后进行转录组分析和功能实验（伤口愈合、WST-1增殖或基质胶实验）以表征SASH1的功能。
SASH1在人血管细胞（HAECs、平滑肌细胞）以及单核细胞/巨噬细胞中表达。与非吸烟者相比，吸烟者动脉粥样硬化颈动脉中的组织表达显著更高（p<0.01）。在HAECs中，SASH1主要在细胞质中表达，SASH1敲低导致细胞迁移、增殖和血管生成增加。转录组和通路分析表明，SASH1沉默可能通过抑制TP53活性导致CYP1A1表达降低。
我们表明吸烟者动脉粥样硬化颈动脉中SASH1表达增加，其沉默影响内皮血管生成功能；因此我们通过SASH1表达提供了吸烟与动脉粥样硬化之间的潜在联系。
