title: oxLDL induces injury and defenestration of human liver sinusoidal endothelial cells via LOX1.
abstract: 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  μg/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), NFκB, p65, (p65), endothelin 1 (ET1 (EDN1)), eNOS (NOS3) and caveolin 1 (CAV1) levels were measured. Cells were treated with 100  μg/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 NFκB activation, upregulated ET1 and caveolin 1 expression, downregulated 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/NFκB signalling pathway and by which LOX1 mediates oxLDL-induced endothelial injury and the defenestration of HLSECs.
authors: The First Clinical College of Lanzhou University730000 Lanzhou, Gansu, ChinaDepartment of EndocrinologyGansu Provincial Hospital, 204 West Donggang Road, 730000 Lanzhou, Gansu, China.
authors: Qi
authors: Q
authors: Zhang
authors: The First Clinical College of Lanzhou University730000 Lanzhou, Gansu, ChinaDepartment of EndocrinologyGansu Provincial Hospital, 204 West Donggang Road, 730000 Lanzhou, Gansu, China liujingwelcome@126.com.
authors: Jing
authors: J
authors: Liu
authors: The First Clinical College of Lanzhou University730000 Lanzhou, Gansu, ChinaDepartment of EndocrinologyGansu Provincial Hospital, 204 West Donggang Road, 730000 Lanzhou, Gansu, China.
authors: Jia
authors: J
authors: Liu
authors: The First Clinical College of Lanzhou University730000 Lanzhou, Gansu, ChinaDepartment of EndocrinologyGansu Provincial Hospital, 204 West Donggang Road, 730000 Lanzhou, Gansu, China.
authors: Wenhui
authors: W
authors: Huang
authors: The First Clinical College of Lanzhou University730000 Lanzhou, Gansu, ChinaDepartment of EndocrinologyGansu Provincial Hospital, 204 West Donggang Road, 730000 Lanzhou, Gansu, China.
authors: Limin
authors: L
authors: Tian
authors: The First Clinical College of Lanzhou University730000 Lanzhou, Gansu, ChinaDepartment of EndocrinologyGansu Provincial Hospital, 204 West Donggang Road, 730000 Lanzhou, Gansu, China.
authors: Jinxing
authors: J
authors: Quan
authors: The First Clinical College of Lanzhou University730000 Lanzhou, Gansu, ChinaDepartment of EndocrinologyGansu Provincial Hospital, 204 West Donggang Road, 730000 Lanzhou, Gansu, China.
authors: Yunfang
authors: Y
authors: Wang
authors: The First Clinical College of Lanzhou University730000 Lanzhou, Gansu, ChinaDepartment of EndocrinologyGansu Provincial Hospital, 204 West Donggang Road, 730000 Lanzhou, Gansu, China.
authors: Ruilan
authors: R
authors: Niu
mesh_terms: D051242:Caveolin 1 / Q000235:genetics / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D019332:Endothelin-1 / Q000235:genetics / Q000378:metabolism; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism* / Q000494:pharmacology / Q000633:toxicity; D008099:Liver / Q000378:metabolism*; D008875:Middle Aged; D016328:NF-kappa B / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D051127:Scavenger Receptors, Class E / Q000235:genetics / Q000378:metabolism*
chemical_list: D051242:Caveolin 1; D019332:Endothelin-1; D008077:Lipoproteins, LDL; D016328:NF-kappa B; C495137:OLR1 protein, human; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D051127:Scavenger Receptors, Class E; C099252:oxidized low density lipoprotein; D052250:Nitric Oxide Synthase Type III
keywords: defenestration; endothelial injury; human liver sinusoidal endothelial cells; lectin-like oxLDL receptor 1; oxidised LDL