Department of Cardiovascular Surgery, Shanghai Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Department of Cardiology, Central Hospital of Minhang District, Shanghai, China.
J Vasc Res. 2023;60(4):204-212. doi: 10.1159/000533320. Epub 2023 Sep 6.
This study aimed to determine the function of Cx43 in the endothelial-to-mesenchymal transition (EndMT) process of endothelial cells (ECs) and to explore the potential signaling pathways underlying these functions.
ECs were extracted from rat aorta. ECs were transfected with Cx43 cDNA and Cx43 siRNA and then exposed to 5 or 12 dyne/cm2. Immunofluorescence staining was used to detect the expression of SM22α, Cx43, and acetylated α-tubulin in ECs. Western blotting was used to detect the protein expression of α-SMA, CD31, Cx43, H1-calponin, Ift88, and p-smad3 in ECs.
The expression of αSMA, SM22α, and Cx43 was significantly increased, and CD31 was markedly decreased in ECs treated with laminar shear stress at 5 dyn/cm2. The Cx43 cDNA transfection could induce the expression of SM22α or H1-calponin and attenuate CD31 expression in ECs. Also, Cx43 overexpression harms cilia formation in ECs exposed to 5 dyn/cm2, accompanied with the regulated Ift88 and smad signaling.
This study found that laminar shear stress at 5 dyn/cm2 would increase the expression of Cx43 to facilitate the EndMT process of ECs, associated with morphological changes in primary cilia and the decreased expression of Ift88 in ECs.
本研究旨在确定缝隙连接蛋白 43（Cx43）在血管内皮细胞（EC）向间充质转化（EndMT）过程中的作用，并探讨其潜在的信号通路。
从大鼠主动脉中提取 EC。将 Cx43 cDNA 和 Cx43 siRNA 转染 EC，然后暴露于 5 或 12 达因/平方厘米。免疫荧光染色用于检测 EC 中 SM22α、Cx43 和乙酰化α-微管蛋白的表达。Western blot 用于检测 EC 中 α-SMA、CD31、Cx43、H1-钙调蛋白、Ift88 和 p-smad3 的蛋白表达。
在 5 达因/平方厘米的层流剪切应力作用下，EC 中 αSMA、SM22α 和 Cx43 的表达显著增加，CD31 明显减少。Cx43 cDNA 转染可诱导 SM22α 或 H1-钙调蛋白的表达，并减弱 EC 中 CD31 的表达。此外，Cx43 过表达会损害 EC 在 5 达因/平方厘米下暴露时的纤毛形成，同时伴随着 Ift88 和 smad 信号的调节。
本研究发现，5 达因/平方厘米的层流剪切应力会增加 Cx43 的表达，促进 EC 的 EndMT 过程，与初级纤毛的形态变化和 EC 中 Ift88 的表达减少有关。
