title: Mitochondrial Reactive Oxygen Species Mediate Lysophosphatidylcholine-Induced Endothelial Cell Activation.
abstract: Hyperlipidemia-induced endothelial cell (EC) activation is considered as an initial event responsible for monocyte recruitment in atherogenesis. However, it remains poorly defined what is the mechanism underlying hyperlipidemia-induced EC activation. Here, we tested a novel hypothesis that mitochondrial reactive oxygen species (mtROS) serve as signaling mediators for EC activation in early atherosclerosis.
  
  
  
  Metabolomics and transcriptomics analyses revealed that several lysophosphatidylcholine (LPC) species, such as 16:0, 18:0, and 18:1, and their processing enzymes, including Pla2g7 and Pla2g4c, were significantly induced in the aortas of apolipoprotein E knockout mice during early atherosclerosis. Using electron spin resonance and flow cytometry, we found that LPC 16:0, 18:0, and 18:1 induced mtROS in primary human aortic ECs, independently of the activities of nicotinamide adenine dinucleotide phosphate oxidase. Mechanistically, using confocal microscopy and Seahorse XF mitochondrial analyzer, we showed that LPC induced mtROS via unique calcium entry-mediated increase of proton leak and mitochondrial O2 reduction. In addition, we found that mtROS contributed to LPC-induced EC activation by regulating nuclear binding of activator protein-1 and inducing intercellular adhesion molecule-1 gene expression in vitro. Furthermore, we showed that mtROS inhibitor MitoTEMPO suppressed EC activation and aortic monocyte recruitment in apolipoprotein E knockout mice using intravital microscopy and flow cytometry methods.
  
  
  
  ATP synthesis-uncoupled, but proton leak-coupled, mtROS increase mediates LPC-induced EC activation during early atherosclerosis. These results indicate that mitochondrial antioxidants are promising therapies for vascular inflammation and cardiovascular diseases.
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Xinyuan
authors.initials: X
authors.lastname: Li
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Pu
authors.initials: P
authors.lastname: Fang
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Yafeng
authors.initials: Y
authors.lastname: Li
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Yin-Ming
authors.initials: YM
authors.lastname: Kuo
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Andrews
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Gayani
authors.initials: G
authors.lastname: Nanayakkara
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Candice
authors.initials: C
authors.lastname: Johnson
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Hangfei
authors.initials: H
authors.lastname: Fu
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Huimin
authors.initials: H
authors.lastname: Shan
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Fuyong
authors.initials: F
authors.lastname: Du
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Nicholas E
authors.initials: NE
authors.lastname: Hoffman
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Daohai
authors.initials: D
authors.lastname: Yu
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Satoru
authors.initials: S
authors.lastname: Eguchi
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Muniswamy
authors.initials: M
authors.lastname: Madesh
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Walter J
authors.initials: WJ
authors.lastname: Koch
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Jianxin
authors.initials: J
authors.lastname: Sun
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Jiang
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.).
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.affiliation: From the Centers for Metabolic Disease Research, Cardiovascular Research, Thrombosis Research (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., S.E., X.J., H.W., X.Y.), Department of Pharmacology (X.L., P.F., Y.L., G.N., C.J., H.F., H.S., F.D., W.J.K., X.J., H.W., X.Y.), Department of Biochemistry (N.E.H., M.M.), Department of Physiology (S.E.), Center for Translational Medicine (N.E.H., M.M., W.J.K.), and Department of Clinical Sciences (D.Y.), Temple University School of Medicine, Philadelphia, PA; Department of Cancer Biology, Fox Chase Cancer Center, Temple University Health System, Philadelphia, PA (Y.-M.K., A.J.A.); and Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA (J.S.). xfyang@temple.edu.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Yang
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D001011:Aorta / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D001018:Aortic Diseases / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D001057:Apolipoproteins E / Q000172:deficiency / Q000235:genetics; D050197:Atherosclerosis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D020013:Calcium Signaling; D002478:Cells, Cultured; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation; D020022:Genetic Predisposition to Disease; D006801:Humans; D006949:Hyperlipidemias / Q000235:genetics / Q000378:metabolism; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D008244:Lysophosphatidylcholines / Q000378:metabolism* / Q000494:pharmacology; D053078:Membrane Potential, Mitochondrial; D055432:Metabolomics / Q000379:methods; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000187:drug effects; D010641:Phenotype; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013997:Time Factors; D018808:Transcription Factor AP-1 / Q000235:genetics / Q000378:metabolism
chemical_list: D000975:Antioxidants; D001057:Apolipoproteins E; C573420:ICAM1 protein, human; D008244:Lysophosphatidylcholines; D017382:Reactive Oxygen Species; D018808:Transcription Factor AP-1; D018799:Intercellular Adhesion Molecule-1
keywords: atherosclerosis; endothelial cell; hyperlipidemia; mitochondria; reactive oxygen species