Department of Cardiology, The First Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China.
Int J Mol Med. 2018 Jan;41(1):33-42. doi: 10.3892/ijmm.2017.3200. Epub 2017 Oct 19.
This study aimed to explore whether angiotensin II (Ang II) inhibits the proliferation of human umbilical vein endothelial cells (HUVECs) by changing mitochondrial energy metabolism, and whether atorvastatin has a protective role via restoration of endothelial function. HUVECs were treated with 1 µM Ang II alone or with 10 µM atorvastatin for 24 h. Proliferation was detected by MTT assay, cell counting, 5‑ethynyl‑2'‑deoxyuridine assay and real‑time cell analyzer. Mitochondrial energy metabolism including oxygen consumption rate and extracellular acidification rate were measured using a Seahorse metabolic flux analyzer. Mitochondrial membrane potential was detected under fluorescence microscope following staining with tetramethylrhodamine. Respiratory chain complexes I‑V were detected using western blotting. The current study showed that Ang II inhibits the proliferation of HUVECs. Results from the Seahorse metabolic flux analyzer indicated that Ang II decreased basal oxygen consumption, maximal respiration capacity, spare respiration capacity, adenosine triphosphate‑linked respiration and non‑mitochondrial respiration. By contrast, Ang II increased the proton leak. Additionally, Ang II increased glycolysis, glycolytic capacity and non‑glycolytic acidification. Furthermore, these effects were all suppressed by atorvastatin. The results indicated that atorvastatin prevents cellular energy metabolism switching from oxidative phosphorylation to glycolysis induced by Ang II and protected the proliferative ability of HUVECs.
本研究旨在探讨血管紧张素 II(Ang II)是否通过改变线粒体能量代谢来抑制人脐静脉内皮细胞(HUVEC)的增殖,以及阿托伐他汀是否通过恢复内皮功能发挥保护作用。将 HUVEC 用 1µM Ang II 单独或用 10µM 阿托伐他汀处理 24 小时。通过 MTT 测定法、细胞计数、5-乙炔基-2'-脱氧尿苷测定法和实时细胞分析仪检测增殖。使用 Seahorse 代谢通量分析仪测量线粒体能量代谢,包括耗氧率和细胞外酸化率。用四甲基罗丹明染色后在荧光显微镜下检测线粒体膜电位。用 Western blot 检测呼吸链复合物 I-V。本研究表明 Ang II 抑制 HUVEC 的增殖。 Seahorse 代谢通量分析仪的结果表明,Ang II 降低了基础耗氧率、最大呼吸能力、备用呼吸能力、三磷酸腺苷相关呼吸和非线粒体呼吸。相比之下,Ang II 增加了质子漏。此外,Ang II 增加了糖酵解、糖酵解能力和非糖酵解酸化。此外,阿托伐他汀抑制了这些作用。结果表明,阿托伐他汀可防止 Ang II 诱导的细胞能量代谢从氧化磷酸化向糖酵解的转变,并保护 HUVEC 的增殖能力。