Rezabakhsh Aysa, Montazersaheb Soheila, Nabat Elahe, Hassanpour Mehdi, Montaseri Azadeh, Malekinejad Hassan, Movassaghpour Ali Akbar, Rahbarghazi Reza, Garjani Alireza
Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran.
Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Bioimpacts. 2017;7(4):219-226. doi: 10.15171/bi.2017.26. Epub 2017 Aug 23.
Under the diabetic condition, sustained production of oxidative/nitrosative stress results in irreversible vascular injuries. A great number of diabetic pathologies, such as inefficient or aberrant neo-angiogenesis emerge following chronic hyperglycemic condition. Lack of enough data exists regarding hydroxychloroquine (HCQ) contribution on angiogenesis during diabetes mellitus. To better address whether HCQ could blunt or exacerbate oxidative status and angiogenesis under high glucose condition (HCG), human umbilical vein endothelial cells (HUVECs) were exposed to 30 µM HCQ in combination with 30 mM glucose over a course of 72 hours. Viability was measured was evaluated by MTT assay. We used Griess method and TBARS assay to monitor changes in the levels of NO and MDA followed by flow cytometric analysis of ROS using DCFDA. To show the impact of HCQ on cell motility and in vitro angiogenic properties, we exploited routine scratch test and in vitro tubulogenesis, respectively. Our data showed that HCQ diminished cell viability under 5 and 30 mM glucose contents. HCQ significantly decreased the total levels of nitric oxide (NO), malondialdehyde (MDA), and reactive oxygen species (ROS) in both sets of environments. Additionally, inhibitory effects were observed on cell migration after exposure to HCQ ( < 0.001). Anti-angiogenic activity of HCQ was confirmed by the reduction of tube areas under a normal or surplus amount of glucose ( < 0.001). In overall, results suggest that HCQ changes the oxidative/nitrosative status of HUVECs both in 5 and 30 mM conditions. HCQ is able to reduce migration and angiogenic activity of HUVECs irrespective of the glucose content.
在糖尿病状态下，氧化/亚硝化应激的持续产生会导致不可逆的血管损伤。在慢性高血糖状态后会出现许多糖尿病病理状况，如低效或异常的新生血管形成。关于羟氯喹（HCQ）在糖尿病期间对血管生成的作用，目前缺乏足够的数据。为了更好地探讨HCQ在高糖条件（HCG）下是否能减轻或加剧氧化状态和血管生成，将人脐静脉内皮细胞（HUVECs）在72小时内暴露于30 μM HCQ与30 mM葡萄糖的组合中。通过MTT法测定细胞活力。我们使用Griess法和硫代巴比妥酸反应物（TBARS）测定法监测一氧化氮（NO）和丙二醛（MDA）水平的变化，随后使用2',7'-二氯二氢荧光素二乙酸酯（DCFDA）通过流式细胞术分析活性氧（ROS）。为了显示HCQ对细胞迁移和体外血管生成特性的影响，我们分别采用了常规划痕试验和体外成管试验。我们的数据表明，在5 mM和30 mM葡萄糖含量下，HCQ降低了细胞活力。在两组环境中，HCQ均显著降低了一氧化氮（NO）、丙二醛（MDA）和活性氧（ROS）的总水平。此外，暴露于HCQ后观察到对细胞迁移有抑制作用（<0.001）。在正常或过量葡萄糖条件下，HCQ通过减少管腔面积证实了其抗血管生成活性（<0.001）。总体而言，结果表明，在5 mM和30 mM条件下，HCQ都会改变HUVECs的氧化/亚硝化状态。无论葡萄糖含量如何，HCQ都能够降低HUVECs的迁移和血管生成活性。
