Department of Cardiology, The Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, PR China; First clinical medicine college of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, PR China.
Department of Central Laboratory, The Affiliated Nanjing Hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, PR China; Department of Clinical Biobank, The Affiliated Nanjing Hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, PR China.
Atherosclerosis. 2019 May;284:110-120. doi: 10.1016/j.atherosclerosis.2019.02.010. Epub 2019 Feb 25.
Oxidative stress-induced endothelial dysfunction is considered to exert a vital role in the development of atherosclerotic coronary heart disease (CHD). NRF2 is a key transcriptional factor against oxidative stress through activation of multiple ARE-mediated genes. Z-Lig is derived from the Ligusticum species with antitumor, anti-inflammation and neuroprotection activities. However, the antioxidant potentials of Z-Lig on endothelial dysfunction and atherosclerosis have not been well elucidated. Therefore, in the present work, we appraise the cytoprotective property and anti-atherosclerosis effect of Z-Lig.
Potential NRF2 activators were screened and verified by luciferase reporter gene assay. The protein and mRNA levels of NRF2 and ARE-mediated genes, and GSH/GSSG level in EA.hy926 cells treated with Z-Lig were detected. The cytoprotective property of Z-Lig was assessed in the tert-butyl hydroperoxide (t-BHP)-evoked oxidative stress model. Cell viability and reactive oxygen species (ROS) levels in EA.hy926 cells were determined. An atherosclerosis model induced by HFD was used to determine the anti-atherosclerosis effect of Z-Lig in HFD-fed Ldlr-deficient mice.
In vitro, 100 μM Z-Lig upregulated expressions of NRF2 and ARE-driven genes, promoted accumulation of nuclear NRF2 and unbound NRF2- KEAP1 complex in EA.hy926 cells. Furthermore, Z-Lig alleviated oxidative stress and cell injury caused by t-BHP via stimulation of the NRF2/ARE pathway. In vivo, intervention with 20 mg/kg Z-Lig markedly restrained atherosclerosis progression, including attenuation of HFD-induced atherosclerotic plaque formation, alleviation of lipid peroxidation and increase in antioxidant enzyme activity in aortas of HFD-fed Ldlr mice. The chemopreventive effects of Z-Lig might be associated with the activation of NRF2 and ARE-driven genes.
The present study suggested that Z-Lig is an effective NRF2 activator, which can protect vascular endothelial cells from oxidative stress and rescue HFD-induced atherosclerosis.
氧化应激诱导的内皮功能障碍被认为在动脉粥样硬化性冠心病（CHD）的发展中发挥重要作用。NRF2 是一种通过激活多个 ARE 介导的基因来对抗氧化应激的关键转录因子。Z-Lig 源自独活属植物，具有抗肿瘤、抗炎和神经保护作用。然而，Z-Lig 对内皮功能障碍和动脉粥样硬化的抗氧化潜力尚未得到充分阐明。因此，在本工作中，我们评估了 Z-Lig 的细胞保护特性和抗动脉粥样硬化作用。
通过荧光素酶报告基因检测筛选和验证潜在的 NRF2 激活剂。检测 Z-Lig 处理的 EA.hy926 细胞中 NRF2 和 ARE 介导基因的蛋白和 mRNA 水平以及 GSH/GSSG 水平。在叔丁基过氧化氢（t-BHP）诱导的氧化应激模型中评估 Z-Lig 的细胞保护特性。测定 EA.hy926 细胞的细胞活力和活性氧（ROS）水平。使用 HFD 诱导的动脉粥样硬化模型确定 Z-Lig 在 HFD 喂养的 Ldlr 缺陷小鼠中的抗动脉粥样硬化作用。
在体外，100μM Z-Lig 上调了 NRF2 和 ARE 驱动基因的表达，促进了 EA.hy926 细胞中核 NRF2 和未结合的 NRF2-KEAP1 复合物的积累。此外，Z-Lig 通过刺激 NRF2/ARE 通路缓解了 t-BHP 引起的氧化应激和细胞损伤。在体内，20mg/kg Z-Lig 的干预显著抑制了动脉粥样硬化的进展，包括减轻 HFD 诱导的动脉粥样硬化斑块形成、减轻脂质过氧化和增加 HFD 喂养的 Ldlr 小鼠主动脉中的抗氧化酶活性。Z-Lig 的化学预防作用可能与 NRF2 和 ARE 驱动基因的激活有关。
本研究表明，Z-Lig 是一种有效的 NRF2 激活剂，可保护血管内皮细胞免受氧化应激，并挽救 HFD 诱导的动脉粥样硬化。
