Department of Pharmacy, Zhujiang Hospital, Southern Medical University, Guangdong, Guangzhou, People's Republic of China.
Affiliated Langdong Hospital, Guangxi Medical University, Guangxi, Nanning, People's Republic of China.
J Cardiovasc Pharmacol. 2018 May;71(5):293-302. doi: 10.1097/FJC.0000000000000571.
The aim of this study is to investigate the potential roles of Rutaecarpine (Rut) in hypoxia-induced human pulmonary artery smooth muscle cells (HPASMCs) model.
HPASMCs were cultured with or without hypoxia followed by Rut administration. Cytotoxicity and cell proliferation were assessed by CCK-8 and Cell counting method. Flow cytometry was used for the measurement of cell apoptosis rates. The mRNA expression of hypoxia-induced factor (HIF)-1α and protein levels of HIF-1α, p53, p21, erythropoietin, and vascular endothelial growth factor were determined by quantitative real-time polymerase chain reaction and Western blot, respectively.
Rut inhibited the proliferation of HPASMCs with IC50 value of 43.5 μmol·L. Hypoxia significantly increased proliferation and decreased apoptosis in HPASMCs, whereas Rut rescued this phenomenon at the appropriate concentration. Meanwhile, Rut effectively decreased the protein and mRNA expressions of HIF-1α. Knockdown of HIF-1α expression by small interfering RNA (siRNA) significantly enhanced the proapoptotic effect rather than antiproliferation effect of Rut in HPASMCs. Moreover, Rut simultaneously reduced proliferating cell nuclear antigen protein expression, whereas increased p53 and p21 protein levels. However, no significant difference was observed in the protein levels of vascular endothelial growth factor and erythropoietin.
Our results demonstrated that Rut exerted protective effects on HPASMCs against hypoxia partly through the HIF-1α-dependent signaling pathway.
本研究旨在探讨吴茱萸碱（Rut）在低氧诱导的人肺动脉平滑肌细胞（HPASMCs）模型中的潜在作用。
在缺氧条件下或缺氧条件下给予 Rut 处理后，培养 HPASMCs。通过 CCK-8 和细胞计数法评估细胞毒性和细胞增殖。通过流式细胞术测定细胞凋亡率。通过定量实时聚合酶链反应和 Western blot 分别测定缺氧诱导因子（HIF）-1α的 mRNA 表达和 HIF-1α、p53、p21、促红细胞生成素和血管内皮生长因子的蛋白水平。
Rut 的 IC50 值为 43.5 μmol·L，抑制 HPASMCs 的增殖。缺氧显著增加 HPASMCs 的增殖并降低其凋亡，而 Rut 在适当浓度下可挽救此现象。同时，Rut 有效降低 HIF-1α的蛋白和 mRNA 表达。用小干扰 RNA（siRNA）敲低 HIF-1α表达显著增强了 Rut 在 HPASMCs 中的促凋亡作用，而非抗增殖作用。此外，Rut 同时降低增殖细胞核抗原蛋白表达，而增加 p53 和 p21 蛋白水平。然而，血管内皮生长因子和促红细胞生成素的蛋白水平没有明显差异。
我们的结果表明，Rut 对 HPASMCs 具有保护作用，可部分通过 HIF-1α 依赖的信号通路来抵抗缺氧。
