Department of Physiology, Harbin Medical University-Daqing, Daqing, China.
Department of Pharmacology, Harbin Medical University-Daqing, Daqing, China.
Cell Prolif. 2018 Jun;51(3):e12436. doi: 10.1111/cpr.12436. Epub 2018 Jan 22.
Transient receptor potential vanilloid 3 (TRPV3) is a member of the TRP channels family of Ca -permeant cation channels. In this study, we aim to investigate the role of TRPV3 in pulmonary vascular remodeling and PASMCs proliferation under hypoxia.
The expression of TRPV3 was evaluated in patients with pulmonary arterial hypertension (PAH) and hypoxic rats, using hematoxylin and eosin (H&E) and immunohistochemistry. In vitro, MTT assay, flow cytometry, Western blotting and immunofluorescence were performed to investigate the effects of TRPV3 on proliferation of PASMCs.
We found that, in vivo, the expression of TRPV3 was increased in patients with PAH and hypoxic rats. Right ventricular hypertrophy measurements and pulmonary pathomorphology data show that the ratio of the heart weight/tibia length (HW/TL), the right ventricle/left ventricle plus septum (RV/LV+S) and the medial width of the pulmonary artery were increased in chronic hypoxic rats. Moreover, the expression of proliferating cell nuclear antigen (PCNA), Cyclin D, Cyclin E and Cyclin A, phospho-CaMKII (p-CaMKII) were induced by hypoxia. In vitro, we revealed that hypoxia promoted PASMCs viability, increased the expression of PCNA, Cyclin D, Cyclin E, Cyclin A p-CaMKII, made more cells from G /G phase to G /M + S phase, enhanced the microtubule formation, and increased [Ca ] , which could be suppressed by Ruthenium Red, an inhibitor of TRPV3, and TRPV3 silencing has similar effects. Furthermore, the up-regulated expression of PCNA, Cyclin D, Cyclin E and Cyclin A, the increased number of cells in G /M and S phase, and the enhanced activation and expression of PI3K and AKT proteins induced by hypoxia and in presence of carvacrol (an agonist of TRPV3), was significantly attenuated by incubation of LY 294002, a specific inhibitor for PI3K/AKT.
These findings suggest that TRPV3 is involved in hypoxia-induced pulmonary vascular remodeling and promotes proliferation of PASMCs and the effect is, at least in part, mediated via the PI3K/AKT pathway.
瞬时受体电位香草酸 3 型（TRPV3）是钙通透性阳离子通道 TRP 通道家族的成员。在这项研究中，我们旨在研究 TRPV3 在低氧环境下肺血管重构和 PASMC 增殖中的作用。
使用苏木精和伊红（H&E）和免疫组织化学评估肺动脉高压（PAH）患者和低氧大鼠中 TRPV3 的表达。在体外，通过 MTT 测定、流式细胞术、Western blot 和免疫荧光研究 TRPV3 对 PASMC 增殖的影响。
我们发现，在体内，PAH 患者和低氧大鼠中 TRPV3 的表达增加。右心室肥厚测量和肺病理形态学数据显示，慢性低氧大鼠的心脏重量/胫骨长度（HW/TL）、右心室/左心室加室间隔（RV/LV+S）和肺动脉中膜宽度比值增加。此外，增殖细胞核抗原（PCNA）、细胞周期蛋白 D、细胞周期蛋白 E 和细胞周期蛋白 A、磷酸化钙调蛋白依赖性蛋白激酶 II（p-CaMKII）的表达受到低氧诱导。在体外，我们发现低氧促进 PASMC 活力，增加 PCNA、细胞周期蛋白 D、细胞周期蛋白 E、细胞周期蛋白 A 和 p-CaMKII 的表达，使更多细胞从 G1/G0 期进入 G2/M+S 期，增强微管形成，增加[Ca2+]，这可被 Ruthenium Red（TRPV3 的抑制剂）抑制，TRPV3 沉默也有类似的作用。此外，低氧和 carvacrol（TRPV3 激动剂）存在下，PCNA、细胞周期蛋白 D、细胞周期蛋白 E 和细胞周期蛋白 A 的上调表达、G2/M 和 S 期细胞数量的增加以及 PI3K 和 AKT 蛋白的激活和表达增强，均被 LY 294002（PI3K/AKT 的特异性抑制剂）孵育显著减弱。
这些发现表明 TRPV3 参与低氧诱导的肺血管重构，并促进 PASMC 的增殖，其作用至少部分是通过 PI3K/AKT 途径介导的。
