Du Qian, Qian Ming-Ming, Liu Pin-Li, Zhang Le, Wang Yan, Liu Dong-Hui
Department of Cardiology, the Affiliated Cardiovascular Hospital of Xiamen University, Medical College of Xiamen University, Xiamen, China.
Department of Endocrinology, the Second Hospital of Hebei Medical University, Shijiazhuang, China.
J Geriatr Cardiol. 2017 Jul;14(7):473-480. doi: 10.11909/j.issn.1671-5411.2017.07.003.
In type 2 diabetes mellitus (T2DM), high-density lipoprotein (HDL) impairs its anti-atherogenic properties and even develops to a pro-inflammatory and pro-atherogenic phenotype because of abnormal compositions and modifications. In this study, we examined the effects and the related mechanisms of glycation of HDL on the proliferation and migration of vascular smooth muscle cells (VSMCs).
METHODS & RESULTS: Glycated HDL (G-HDL) was modified with D-glucose (25 mmol/L) . Diabetic HDL (D-HDL) was isolated from T2DM patients. Rat VSMCs were isolated from the thoracic aortas. Human VSMCs were obtained from ScienCell Research Laboratories. Alpha-actin was detected through immunofluorescence. VSMC proliferation was assayed by Cell Count. VSMC migration was determined by transwell chamber and scratch-wound assay. Intracellular reactive oxygen species (ROS) was detected based on ROS-mediated 2',7'-dichlorofluorescein (DCFH-DA) fluorescence. Compared to native HDL (N-HDL), G-HDL remarkably promoted VSMC proliferation and migration in the dose and time-dependent manners. In addition, G-HDL enhanced ROS generation in VSMCs. However, the ROS scavenger, N-acetylcysteine, efficiently decreased ROS production and subsequently inhibited the proliferation of VSMCs induced by G-HDL. Similarly, D-HDL from T2DM patients also promoted ROS release and VSMC proliferation and migration.
HDL either glycated or isolated from T2DM patients triggered VSMC proliferation, migration, and oxidative stress. These results might partly interpret the higher morbidity of cardiovascular disease in T2DM patients.
在2型糖尿病（T2DM）中，高密度脂蛋白（HDL）因其组成和修饰异常而损害其抗动脉粥样硬化特性，甚至发展为促炎和促动脉粥样硬化表型。在本研究中，我们研究了HDL糖基化对血管平滑肌细胞（VSMC）增殖和迁移的影响及其相关机制。
用D-葡萄糖（25 mmol/L）修饰糖基化HDL（G-HDL）。从T2DM患者中分离出糖尿病HDL（D-HDL）。从胸主动脉分离大鼠VSMC。人VSMC购自ScienCell Research Laboratories。通过免疫荧光检测α-肌动蛋白。通过细胞计数测定VSMC增殖。通过Transwell小室和划痕试验测定VSMC迁移。基于活性氧（ROS）介导的2',7'-二氯荧光素（DCFH-DA）荧光检测细胞内ROS。与天然HDL（N-HDL）相比，G-HDL以剂量和时间依赖性方式显著促进VSMC增殖和迁移。此外，G-HDL增强了VSMC中的ROS生成。然而，ROS清除剂N-乙酰半胱氨酸有效地降低了ROS产生，并随后抑制了G-HDL诱导的VSMC增殖。同样，来自T2DM患者的D-HDL也促进了ROS释放以及VSMC增殖和迁移。
糖基化或从T2DM患者中分离出的HDL均可引发VSMC增殖、迁移和氧化应激。这些结果可能部分解释了T2DM患者心血管疾病发病率较高的原因。
