Department of Respiratory and Critical Care Medicine, Key Laboratory of Pulmonary Diseases of Health Ministry, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Department of Respiratory and Critical Care Medicine, Affiliated Hospital of Zunyi Medical University, Zunyi, China.
J Cell Mol Med. 2021 Jan;25(2):1238-1251. doi: 10.1111/jcmm.16194. Epub 2020 Dec 20.
We found previously that KLF4 expression was up-regulated in cultured rat and human pulmonary artery smooth muscle cells (PASMCs) exposed to cigarette smoke (CS) extract and in pulmonary artery from rats with pulmonary hypertension induced by CS. Here, we aim to investigate whether CS-induced pulmonary hypertension (PH) is prevented and ameliorated by targeted pulmonary vascular gene knockdown of KLF4 via adeno-associated virus 1 (AAV1)-KLF4-shRNA in vivo in rat model. The preventive and therapeutic effects were observed according to the different time-point of AAV1-KLF4-shRNA intratracheal administration. We tested haemodynamic measurements of systemic and pulmonary circulations and observed the degree of pulmonary vascular remodelling. In the preventive experiment, KLF4 expression and some pulmonary circulation hemodynamic measurements such as right ventricular systolic pressure (RVSP), mean right ventricular pressure (mRVP), peak RV pressure rate of rise (dP/dt max) and right ventricle (RV) contractility index were increased significantly in the CS-induced PH model. While in the prevention group (AAV1-KLF4-shRNA group), RVSP, mRVP, dP/dt max and RV contractility index which are associated with systolic function of right ventricle decreased and the degree of pulmonary vascular remodelling relieved. In the therapeutic experiment, we observed a similar trend. Our findings emphasize the feasibility of sustained pulmonary vascular KLF4 gene knockdown using intratracheal delivery of AAV1 in an animal model of cigarette smoke-induced PH and determined gene transfer of KLF4-shRNA could prevent and ameliorate the progression of PH.
我们之前发现，在培养的大鼠和人肺动脉平滑肌细胞（PASMCs）中，在接触香烟烟雾（CS）提取物以及在 CS 诱导的肺动脉高压大鼠的肺动脉中，KLF4 的表达上调。在这里，我们旨在通过腺相关病毒 1（AAV1）-KLF4-shRNA 在体内大鼠模型中靶向肺血管基因敲低 KLF4，来研究 CS 诱导的肺动脉高压（PH）是否可以预防和改善。根据 AAV1-KLF4-shRNA 经气管内给药的不同时间点观察预防和治疗效果。我们测试了体循环和肺循环的血流动力学测量值，并观察了肺血管重构的程度。在预防实验中，在 CS 诱导的 PH 模型中，KLF4 的表达和一些肺循环血流动力学测量值，如右心室收缩压（RVSP）、平均右心室压（mRVP）、右心室收缩压上升率（dP/dt max）和右心室（RV）收缩性指数，均显著增加。而在预防组（AAV1-KLF4-shRNA 组）中，与右心室收缩功能相关的 RVSP、mRVP、dP/dt max 和 RV 收缩性指数降低，肺血管重构的程度得到缓解。在治疗实验中，我们观察到了类似的趋势。我们的研究结果强调了使用气管内给予 AAV1 持续肺血管 KLF4 基因敲低在 CS 诱导的 PH 动物模型中的可行性，并确定 KLF4-shRNA 的基因转移可以预防和改善 PH 的进展。
