Almazov Federal Medical Research Centre, Saint-Petersburg, Russia; Institute of Cytology, Russian Academy of Sciences, Saint-Petersburg, Russia.
Almazov Federal Medical Research Centre, Saint-Petersburg, Russia; Institute of Cytology, Russian Academy of Sciences, Saint-Petersburg, Russia; Saint-Petersburg State University, Saint-Petersburg, Russia.
Biochem Biophys Res Commun. 2019 Jun 25;514(2):462-468. doi: 10.1016/j.bbrc.2019.04.177. Epub 2019 May 2.
Cardiovascular calcification is one of the leading reasons of morbidity and mortality in Western countries and has many similarities to osteogenesis. The role of smooth muscle calcific transformation is well established for atherogenic lesions, but mechanisms driving initial stages of proosteogenic cell fate commitment in big vessels remain poorly understood. The role of endothelial and underlying interstitial cell interaction in driving cellular decisions is emerging from recent studies. The aim of this study was to analyze co-culture of endothelial and smooth muscle cells in vitro in acquiring proosteogenic phenotype. We co-cultured human aortic endothelial cells (EC) and human aortic smooth muscle cells (SMC) and analyzed osteogenic phenotype by ALP staining and proosteogenic gene expression by qPCR in co-cultures and in separate cellular types after magnetic CD31-sorting. In EC and SMC co-cultures osteogenic phenotype was induced as well as activated expression of RUNX2, POSTIN, BMP2/4, SOX5, COL1A SMC; co-culture of EC with SMC induced NOTCH1, NOTCH3, NOTCH4 and HEY1 expression; Notch activation by lentiviral activated Notch intracellular domain induced expression of RUNX2, OPN, POSTIN in SMC; NOTCH1 and NOTCH3 and HEY1 were selectively induced in EC during co-culture. We conclude that endothelial cells are capable of driving smooth muscle calcification via cell-cell contact and activation of Notch signaling.
心血管钙化是西方国家发病率和死亡率的主要原因之一，与成骨有许多相似之处。平滑肌钙化转化在动脉粥样硬化病变中作用明确，但大血管中初始促成骨细胞命运决定的机制仍知之甚少。内皮细胞和下间质细胞相互作用在驱动细胞决策中的作用正从最近的研究中显现出来。本研究旨在分析体外共培养内皮细胞和平滑肌细胞以获得成骨表型。我们共培养人主动脉内皮细胞（EC）和人主动脉平滑肌细胞（SMC），通过碱性磷酸酶染色分析成骨表型，并通过 qPCR 分析共培养物和经磁 CD31 分选后的单独细胞类型中成骨基因的表达。在 EC 和 SMC 的共培养物中诱导了成骨表型，同时 RUNX2、POSTIN、BMP2/4、SOX5、COL1A SMC 的激活表达；EC 与 SMC 的共培养诱导了 NOTCH1、NOTCH3、NOTCH4 和 HEY1 的表达；慢病毒激活 Notch 细胞内结构域诱导 SMC 中 RUNX2、OPN、POSTIN 的表达；共培养过程中，NOTCH1、NOTCH3 和 HEY1 在内皮细胞中选择性诱导。我们得出结论，内皮细胞通过细胞间接触和 Notch 信号的激活能够驱动平滑肌钙化。
