AAV1-Mediated shRNA Knockdown of SASH1 in Rat Bronchus Attenuates Hypoxia-Induced Pulmonary Artery Remodeling
AAV1-Mediated shRNA Knockdown of SASH1 in Rat Bronchus Attenuates Hypoxia-Induced Pulmonary Artery Remodeling
复制标题
AAV1 介导的大鼠支气管中 SASH1 的 shRNA 敲低可减轻缺氧引起的肺动脉重塑
DOI：
10.1089/hum.2020.242
复制
发表时间：
2021-02-22
影响因子：
4.2
通讯作者：
Li, Qinghai
中科院分区：
文献类型：
--
作者：
Liu, Hong;Wang, Ning;Li, Qinghai
肺动脉高压（PH）是一种以肺动脉重构（PAR）为特征的增生性疾病。SAM and SH 3 domain containing 1（SASH 1）是一种新型肿瘤抑制基因，其在PH中的生物学功能尚不清楚。本研究通过建立低氧诱导的肺动脉高压（HPH）大鼠模型，探讨SASH 1在PAR中的作用。通过腺相关病毒1型（AAV 1）感染SASH 1基因敲除大鼠，观察肺组织病理学变化和血流动力学改变。体外培养人肺动脉平滑肌细胞（HPASMCs），用SASH 1 siRNA转染HPASMCs，观察SASH 1对缺氧诱导的HPASMCs增殖和迁移的影响。SASH 1相关的分子机制通过敲低和过表达的方法进行了探索。我们发现，SASH 1的表达显着增加大鼠肺动脉和HPASMCs缺氧暴露后。在体内，沉默SASH 1基因表达改善了大鼠的HPH。SASH 1下调可抑制缺氧诱导的HPASMCs增殖和迁移。SASH 1过表达后，HPASMCs中磷酸化AKT（称为蛋白激酶B）、增殖细胞核抗原和基质金属蛋白酶9（MMP 9）的蛋白表达增加，而这些作用被SASH 1敲低抑制。总之，SASH1下调改善缺氧诱导的PAR和PH。SASH1可能是精准医学时代PH基因治疗的新靶点。
Pulmonary hypertension (PH) is a proliferative disease characterized by pulmonary arterial remodeling (PAR). SAM and SH3 domain containing 1 (SASH1) is a novel tumor suppressor gene whose biological function in PH is unclear. In this study, a hypoxia-induced pulmonary hypertension (HPH) rat model was constructed to explore the role of SASH1 in PAR. Histopathological changes in the lung tissue and hemodynamic alteration were detected in SASH1-knockdown rats through adeno-associated virus type-1 (AAV1) infection. In vitro, primary human pulmonary arterial smooth muscle cells (HPASMCs) were transfected with SASH1siRNA to investigate the effects of SASH1 on hypoxia-induced proliferation and migration. The molecular mechanisms associated with SASH1 were explored through knockdown and overexpression approaches. We found that SASH1 expression was significantly increased in rat pulmonary arteries and HPASMCs after hypoxia exposure. In vivo, silencing the SASH1 gene expression improved HPH in rats. The SASH1 downregulation inhibited proliferation and migration of hypoxia-induced HPASMCs. The protein expression of phospho-AKT (known as protein kinase B), proliferating cell nuclear antigen, and matrix metalloproteinase 9 (MMP9) in HPASMCs were increased after SASH1 overexpression, whereas these effects were inhibited by SASH1 knockdown. In conclusion, SASH1 downregulation improved hypoxia-induced PAR and PH. SASH1 may be a novel target for PH gene therapy in the era of precision medicine.