Yao Zhu1, Yajie Zhang2, Xia Huang3, Yong Xie1, Yuan Qu3, Hongyan Long2, Ning Gu4, Weimin Jiang5. 1. 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. 2. 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. 3. Nanjing University of Chinese Medicine, Nanjing, Jiangsu, PR China; Department of Cardiology, The Affiliated Nanjing Hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, PR China. 4. Department of Cardiology, The Affiliated Nanjing Hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, PR China. Electronic address: jsguning@163.com. 5. 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. Electronic address: jwm0410@163.com.
Abstract
BACKGROUND AND AIMS: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.
BACKGROUND AND AIMS: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.