Department of General Surgery II, Central Hospital of Luohe, Luohe, Henan 462000, P.R. China.
Department of Vascular Surgery, The Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, P.R. China.
Int J Mol Med. 2019 Feb;43(2):839-849. doi: 10.3892/ijmm.2018.3996. Epub 2018 Nov 26.
In the present study, the function of microRNA (miR)‑140‑5p on oxidative stress in mice with atherosclerosis was investigated. A reverse transcription‑quantitative polymerase chain reaction assay was used to determine the expression of miR‑140‑5p. Oxidative stress kits and reactive oxygen species (ROS) kits were used to analyze alterations in oxidative stress and ROS levels. The alterations in protein expression were determined using western blot analysis and an immunofluorescence assay. miR‑140‑5p expression was increased in mice with atherosclerosis with hypertension. Consistently, miR‑140‑5p expression was also increased in mice with atherosclerosis. Upregulation of miR‑140‑5p increased oxidative stress and ROS levels by suppressing the protein expression of nuclear factor erythroid 2‑related factor 2 (Nrf2), sirtuin 2 (Sirt2), Kelch‑like enoyl‑CoA hydratase‑associated protein 1 (Keap1) and heme oxygenase 1 (HO‑1) in vitro. By contrast, downregulation of miR‑140‑5p decreased oxidative stress and ROS levels by activating the protein expression of Nrf2, Sirt2, Keap1 and HO‑1 in vitro. Sirt2 agonist or Nrf2 agonist inhibited the effects of miR‑140‑5p on oxidative stress in vitro. Collectively, these results suggested that miR‑140‑5p aggravated hypertension and oxidative stress of mice with atherosclerosis by targeting Nrf2 and Sirt2.
在本研究中,研究了 microRNA(miR)-140-5p 对动脉粥样硬化小鼠氧化应激的作用。采用逆转录-定量聚合酶链反应(qPCR)检测 miR-140-5p 的表达。采用氧化应激试剂盒和活性氧(ROS)试剂盒分析氧化应激和 ROS 水平的变化。采用 Western blot 分析和免疫荧光测定法检测蛋白表达的变化。高血压合并动脉粥样硬化小鼠的 miR-140-5p 表达增加。一致地,动脉粥样硬化小鼠的 miR-140-5p 表达也增加。miR-140-5p 的上调通过抑制核因子红细胞 2 相关因子 2(Nrf2)、沉默调节蛋白 2(Sirt2)、Kelch 样烯酰辅酶 A 水合酶相关蛋白 1(Keap1)和血红素加氧酶 1(HO-1)的蛋白表达,增加了体外的氧化应激和 ROS 水平。相比之下,miR-140-5p 的下调通过激活 Nrf2、Sirt2、Keap1 和 HO-1 的蛋白表达,降低了体外的氧化应激和 ROS 水平。Sirt2 激动剂或 Nrf2 激动剂抑制了 miR-140-5p 对体外氧化应激的作用。综上所述,这些结果表明,miR-140-5p 通过靶向 Nrf2 和 Sirt2 加重了动脉粥样硬化小鼠的高血压和氧化应激。