School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, No.24,Tong Jia Xiang, Nanjing 210009, PR China; Department of Pharmacy, The First People's Hospital of Lianyungang, No.182,TongguanNorth Road, Lianyungang 222002, PR China.
Anesthesiology Department, Children's Hospital of Soochow University, Suzhou 215025, China.
Vascul Pharmacol. 2019 Sep;120:106568. doi: 10.1016/j.vph.2019.106568. Epub 2019 Jun 14.
Pulmonary arterial hypertension (PAH) is a life-threatening disease characterized by the vascular remodeling that also involves proliferation and migration of pulmonary artery smooth muscle cells (PASMCs). Overexpression of Storkhead box (STOX1) regulates genes involved hypoxia, redox balance, nitric oxide, and energy metabolism. In this study, we supposed Stox1 adjusted cells proliferation and migration in PASMCs development and played an important role in the pulmonary arterial vascular remodeling.
Hemodynamic assay and Right ventricular morphometric assay were used to check the rat model of PAH. HE staining was used to examine the arterial wall thickness. Masson staining showed that the deposition of collagen was significantly increased in PAH. In addition, Stox1 were assessed by immunofluorescence and immunohistochemistry staining. The effect of Stox1 on PASMCs was assessed by cell counting Kit-8 assay (CCK-8 assay), Scratch-Wound assay, EdU staining assay, Cell cycle analysis and Western blot.
Right ventricular systolic pressure (RVSP) and right ventricular were significantly increased in hypoxia group and monocrotaline group compared to control group. The expression of Stox1 was increased in lung tissues in PAH rats. In vitro, the expression of Stox1 was up-regulated with time-dependent manner in hypoxia condition. Meanwhile, Stxo1 promoted the proliferation and migration in hypoxia-treated PASMCs. Moreover, we found that hypoxia promoted the expression of PCNA, Cyclin E and Cyclin A, increased more cells from G/G phase to S phase and induced the activation of AKT proteins, which was significantly attenuated by inhibition of Stox1 expression in PASMCs.
These findings indicated that Stox1 induced proliferation of PASMCs and the effect is, at least in part, mediated through AKT signaling pathway.
肺动脉高压（PAH）是一种危及生命的疾病，其特征是血管重构，还涉及肺动脉平滑肌细胞（PASMC）的增殖和迁移。Storkhead 盒（STOX1）的过表达调节涉及缺氧、氧化还原平衡、一氧化氮和能量代谢的基因。在这项研究中，我们假设 Stox1 调节 PASMCs 发育中的细胞增殖和迁移，并在肺血管重构中发挥重要作用。
使用血流动力学测定和右心室形态计量学测定检查 PAH 大鼠模型。HE 染色用于检查动脉壁厚度。Masson 染色显示 PAH 中胶原沉积明显增加。此外，通过免疫荧光和免疫组织化学染色评估 Stox1。通过细胞计数试剂盒-8 测定（CCK-8 测定）、划痕愈合测定、EdU 染色测定、细胞周期分析和 Western blot 评估 Stox1 对 PASMCs 的影响。
与对照组相比，缺氧组和单克隆抗体组的右心室收缩压（RVSP）和右心室均显著升高。PAH 大鼠肺组织中 Stox1 的表达增加。在体外，缺氧条件下 Stox1 的表达随时间呈时间依赖性增加。同时，Stxo1 促进了缺氧处理的 PASMCs 的增殖和迁移。此外，我们发现缺氧促进了 PCNA、Cyclin E 和 Cyclin A 的表达，使更多的细胞从 G/G 期进入 S 期，并诱导 AKT 蛋白的激活，这在 PASMCs 中抑制 Stox1 表达后明显减弱。
这些发现表明 Stox1 诱导 PASMCs 的增殖，其作用至少部分通过 AKT 信号通路介导。
