Guo Yansong, Li Wei, Qian Mingming, Jiang Ting, Guo Ping, Du Qian, Lin Na, Xie Xianwei, Wu Zhiyong, Lin Donghai, Liu Donghui
Department of Cardiology, Fujian Provincial Hospital, Fujian Provincial Key Laboratory of Cardiovascular Disease, Fujian Cardiovascular Institute, Fujian Provincial Center for Geriatrics, Provincial Clinical Medicine College of Fujian Medical University, Fuzhou, China.
Department of Cardiology, the Affiliated Xiamen Cardiovascular Hospital of Xiamen University, Medical College of Xiamen University, Xiamen, China.
Front Pharmacol. 2021 Feb 15;11:556074. doi: 10.3389/fphar.2020.556074. eCollection 2020.
Endothelial dysfunction is involved in the pathophysiological processes of contrast media (CM)-induced acute kidney injury (CI-AKI) after vascular angiography or intervention. Previous study found that apolipoprotein A-I (apoA-I) mimetic peptide, D-4F, alleviates endothelial impairments via upregulating heme oxygenase-1 (HO-1) expression and scavenging excessively generated reactive oxygen species (ROS). However, whether D-4F could ameliorate oxidative injuries in endothelial cells through suppressing ROS production remains unclear. In this study, a representative nonionic iodinated CM, iodixanol, was chosen for the and studies. Endothelial cell viability was assayed using micrographs, lactate dehydrogenase (LDH) activity, and cell counting kit-8 (CCK-8). Apoptosis was detected using flow cytometry analysis and caspase-3 activation. Endothelial inflammation was tested using monocyte adhesion assay and adhesion molecule expression. ROS production was detected by measuring the formation of lipid peroxidation malondialdehyde (MDA) through the thiobarbituric acid reactive substance (TBARS) assay. Peroxynitrite (ONOO⁻) formation was tested using the 3-nitrotyrosine ELISA kit. Iodixanol impaired cell viability, promoted vascular cell adhesion molecule-1 (VCAM-1) and intercellular cell adhesion molecule-1 (ICAM-1) expression, and induced cell apoptosis in human umbilical vein endothelial cells (HUVECs). However, D-4F mitigated these injuries. Furthermore, iodixanol induced the phosphorylation of protein kinase C (PKC) beta II, p47, Rac1, and endothelial nitric oxide synthase (eNOS) at Thr495, which elicited ROS release and ONOO⁻ generation. D-4F inhibited NADPH oxidase (NOX) activation, ROS production, and ONOO⁻ formation via the AMP-activated protein kinase (AMPK)/PKC pathway. Additionally, after an intravascular injection of iodixanol in Sprague Dawley rats, iodixanol induced a remarkable inflammatory response in arterial endothelial cells, although significant apoptosis and morphological changes were not observed. D-4F alleviated the vessel inflammation resulting from iodixanol . Collectively, besides scavenging ROS, D-4F could also suppress ROS production and ONOO⁻ formation through the AMPK/PKC pathway, which ameliorated oxidative injuries in endothelial cells. Hence, D-4F might serve as a potential agent in preventing CI-AKI.
内皮功能障碍参与了血管造影或介入术后造影剂（CM）诱导的急性肾损伤（CI-AKI）的病理生理过程。先前的研究发现，载脂蛋白A-I（apoA-I）模拟肽D-4F通过上调血红素加氧酶-1（HO-1）表达和清除过量产生的活性氧（ROS）来减轻内皮损伤。然而，D-4F是否能通过抑制ROS产生来改善内皮细胞的氧化损伤仍不清楚。在本研究中，选用了一种代表性的非离子型碘化CM碘克沙醇进行体内和体外研究。使用显微镜图像、乳酸脱氢酶（LDH）活性和细胞计数试剂盒-8（CCK-8）检测内皮细胞活力。使用流式细胞术分析和半胱天冬酶-3激活检测细胞凋亡。使用单核细胞黏附试验和黏附分子表达检测内皮炎症。通过硫代巴比妥酸反应物质（TBARS）试验测量脂质过氧化丙二醛（MDA）的形成来检测ROS产生。使用3-硝基酪氨酸ELISA试剂盒检测过氧亚硝酸盐（ONOO⁻）的形成。碘克沙醇损害细胞活力，促进血管细胞黏附分子-1（VCAM-1）和细胞间黏附分子-1（ICAM-1）表达，并诱导人脐静脉内皮细胞（HUVECs）凋亡。然而，D-4F减轻了这些损伤。此外，碘克沙醇诱导蛋白激酶C（PKC）βII、p47、Rac1和内皮型一氧化氮合酶（eNOS）在Thr495位点的磷酸化，从而引发ROS释放和ONOO⁻生成。D-4F通过AMP激活的蛋白激酶（AMPK）/PKC途径抑制NADPH氧化酶（NOX）激活、ROS产生和ONOO⁻形成。此外，在对Sprague Dawley大鼠进行血管内注射碘克沙醇后，碘克沙醇在动脉内皮细胞中诱导了显著的炎症反应，尽管未观察到明显的细胞凋亡和形态学变化。D-4F减轻了碘克沙醇引起的血管炎症。总体而言，除了清除ROS外，D-4F还可通过AMPK/PKC途径抑制ROS产生和ONOO⁻形成，从而改善内皮细胞的氧化损伤。因此，D-4F可能是预防CI-AKI的潜在药物。
