oxLDL induces injury and defenestration of human liver sinusoidal endothelial cells via LOX1
oxLDL induces injury and defenestration of human liver sinusoidal endothelial cells via LOX1
复制标题
OxLDL通过LOX1诱导人肝窦内皮细胞损伤和脱窗
DOI：
10.1530/jme-14-0049
复制
发表时间：
2014-10-01
影响因子：
3.5
通讯作者：
Niu, Ruilan
中科院分区：
文献类型：
--
作者：
Zhang, Qi;Liu, Jing;Niu, Ruilan
非酒精性脂肪肝与2型糖尿病引起的肝脏微血管病变和肝脏炎症有关。氧化低密度脂蛋白（oxLDL）参与各种微循环系统中的促炎症和细胞毒性事件。凝集素样oxLDL受体1（LOX 1）在oxLDL诱导的病理转化中起着至关重要的作用。然而，氧化低密度脂蛋白对肝脏微循环障碍的影响的潜在机制仍不清楚。在这项研究中，我们研究了oxLDL对LOX 1（OLR 1）表达和功能以及体外人肝窦内皮细胞（HLSEC）开窗特征的影响。获得并培养原代HLSEC。用不同浓度的oxLDL（25、50、100和200 μ g/ml）处理细胞，并检查细胞毒性和LOX 1的表达。采用siRNA技术敲除LOX 1，检测细胞内活性氧（ROS）、NF κ B B、p65、内皮素1（ET 1（EDN 1））、eNOS 3（NOS 3）和窖蛋白1（CAV 1）水平的变化。用100 μ g/ml oxLDL处理细胞，并使用扫描电子显微镜观察窗孔形态。oxLDL以剂量和时间依赖性方式显著增加HLSEC中LOX 1在mRNA和蛋白水平的表达。oxLDL刺激增加ROS生成和NF κ B B活化，上调ET 1和caveolin 1表达，下调eNOS表达，降低窗孔直径和孔隙度。所有这些oxLDL介导的作用在LOX 1敲低后被抑制。这些结果揭示了oxLDL通过ROS/NF κ B信号通路刺激LOX 1产生的机制，以及LOX 1通过该机制介导oxLDL诱导的内皮损伤和HLSEC的出窗。
Non-alcoholic fatty liver disease is associated with hepatic microangiopathy and liver inflammation caused by type 2 diabetes mellitus. Oxidised LDL (oxLDL) is involved in proinflammatory and cytotoxic events in various microcirculatory systems. The lectin-like oxLDL receptor 1 (LOX1) plays a crucial role in oxLDL-induced pathological transformation. However, the underlying mechanism of oxLDL's effects on liver microcirculation disturbances remains unclear. In this study, we investigated the effects of oxLDL on LOX1 (OLR1) expression and function, as well as on the fenestration features of human liver sinusoidal endothelial cells (HLSECs) in vitro. Primary HLSECs were obtained and cultured. The cells were treated with various concentrations of oxLDL (25, 50, 100 and 200 mu g/ml), and the cytotoxicity and expression of LOX1 were examined. Furthermore, LOX1 knockdown was performed using siRNA technology, and the changes in intracellular reactive oxygen species (ROS), NF kappa B, p65, (p65), endothelin 1 (ET1 (EDN1)), eNOS (NOS3) and caveolin 1 (CAV1) levels were measured. Cells were treated with 100 mu g/ml oxLDL, and the fenestra morphology was visualised using scanning electron microscopy. oxLDL significantly increased LOX1 expression at both the mRNA and protein levels in HLSECs in a dose-and time-dependent manner. oxLDL stimulation increased ROS generation and NF kappa B activation, upregulated ET1 and caveolin 1 expression, down-regulated eNOS expression and reduced the fenestra diameter and porosity. All of these oxLDL-mediated effects were inhibited after LOX1 knockdown. These results reveal a mechanism by which oxLDL stimulates the production of LOX1 through the ROS/NF kappa B signalling pathway and by which LOX1 mediates oxLDL-induced endothelial injury and the defenestration of HLSECs.