Hu Qingsong, Zhang Baojian, Liu Yulong, Guo Yiqun, Zhang Tao, Nie Ruqiong, Ke Xiao, Dong Xiaobian
Department of Cardiology, First Affiliated Hospital of Jinan University, Guangzhou, China.
Cardiac Care Unit, Affiliated Hospital of Traditional Chinese Medicine, Xinjiang Medical University, Urumqi, China.
Ann Transl Med. 2020 Oct;8(20):1318. doi: 10.21037/atm-20-6467.
Physiological fluid shear stress has been shown to have a beneficial impact on vascular homeostasis. Endothelial progenitor cells (EPCs) make a significant contribution to maintaining endothelial integrity. Therefore, we hypothesised that shear stress-induced endothelium protection plays a role in hydrogen sulphide (HS) production and up-regulation of cystathionine γ-lyase (CSE) expression in EPCs.
Human EPC-derived CSE activity was detected by colorimetric assay, and HS production was evaluated by membrane adsorption method. Cell proliferation, migration, and adhesion were assessed by MTT, Transwell, and endothelial cell-mediated adhesion assays, respectively. Real-time polymerase chain reaction (RT-PCR) was carried out to analyse gene expression. Protein expression was analysed by western blot.
Human EPCs were treated with shear stress levels of 5-25 dyn/cm for up to 3 h, and 25 dyn/cm for up to 24 h. HS production and CSE mRNA expression in the EPCs were increased by shear stress in a dose-dependent manner . Likewise, time-dependent shear stress also significantly enhanced CSE protein expression. Compared to static condition, shear stress improved EPCs proliferation, migration and adhesion capacity. Knockdown of CSE expression by small interfering RNA substantially eliminated the shear stress-induced above functions of human EPCs .
This study gives new insight into the regulatory effect of physiological shear stress on the CSE/HS system in human EPCs. Our findings may contribute to the development of vascular protective research, although the relevant evidence is admittedly indirect.
生理流体剪切应力已被证明对血管稳态有有益影响。内皮祖细胞(EPCs)对维持内皮完整性有重要贡献。因此,我们假设剪切应力诱导的内皮保护在EPCs中硫化氢(HS)产生和胱硫醚γ-裂解酶(CSE)表达上调中起作用。
采用比色法检测人EPCs衍生的CSE活性,采用膜吸附法评估HS产生。分别通过MTT、Transwell和内皮细胞介导的黏附试验评估细胞增殖、迁移和黏附。进行实时聚合酶链反应(RT-PCR)分析基因表达。通过蛋白质印迹分析蛋白质表达。
将人EPCs用5-25达因/平方厘米的剪切应力处理长达3小时,以及用25达因/平方厘米处理长达24小时。EPCs中HS产生和CSE mRNA表达随剪切应力呈剂量依赖性增加。同样,时间依赖性剪切应力也显著增强CSE蛋白表达。与静态条件相比,剪切应力改善了EPCs的增殖、迁移和黏附能力。用小干扰RNA敲低CSE表达基本上消除了剪切应力诱导的人EPCs的上述功能。
本研究为生理剪切应力对人EPCs中CSE/HS系统的调节作用提供了新的见解。我们的发现可能有助于血管保护研究的发展,尽管相关证据诚然是间接的。