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Metformin improved oxidized low-density lipoprotein-impaired mitochondrial function and increased glucose uptake involving Akt-AS160 pathway in raw264.7 macrophages

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Author:: No author available

Journal Title:: Chinese Medical Journal

Issue:: 14

DOI:: 10.1097/CM9.0000000000000333

Key Word:: Atherosclerosis;Macrophages;Oxidized low-density lipoprotein;Mitochondria;Metabolism

Abstract： Background:Macrophage accumulation in the vascular wall is a hallmark of atherosclerosis.Studies showed that shifting of oxidized lipids-induced inflammatory macrophages towards an anti-inflammatory phenotype by promoting oxidative metabolism attenuated atherosclerosis progression.Therefore,this study aimed to investigate whether metformin,which has ameliorated atherosclerosis in animal models and clinical trials,modulated oxidized low-density lipoprotein (Ox-LDL) induced inflammatory status in macrophages by regulating cellular oxidative metabolism.Methods:Murine raw264.7 macrophages were incubated with Ox-LDL (50 μg/mL) in the presence or absence of metformin (15 μmol/L) for 24 h.Real-time polymerase chain reaction was used to quantify the transcription of classically activated (M1) proinflammatory and alternatively activated (M2) anti-inflammatory markers and mitochondrial DNA copy numbers.Cellular reactive oxygen species (ROS) production and mitochondrial membrane potential were detected by immunofluorescence.Cellular adenosine triphosphate (ATP) synthesis,glucose uptake,and lactic acid production were measured by commercial kit and normalized to cellular lysates.Western blotting analysis was performed to detect the expression of mitochondrial fusion/fission related proteins,enzymes mediating lipid metabolism and signaling pathway of glucose transport.Differences between groups were analyzed using one-way analysis of variance.Results:Metformin improved Ox-LDL-impaired anti-inflammatory phenotype in raw264.7 macrophages as shown by up-regulated transcription of anti-inflammatory markers including interleukin 10 (0.76 ± 0.04 vs.0.94 ± 0.01,P =0.003) and Resistin-like molecule alpha (0.67 ± 0.08 vs.1.78 ± 0.34,P =0.030).Conversely,Ox-LDL-diminished phosphorylation of Akt was up regulated by metformin treatment (0.47 ± 0.05 vs.1.02 ± 0.08,P =0.040),associated with an improvement of mitochondrial function,characterized by decreased ROS generation (2.50 ± 0.07 vs.2.15 ± 0.04,P =0.040),increased lipid oxidation,and elevated cellular ATP production (0.026 ± 0.001 vs.0.035 ± 0.003,P =0.020).Moreover,metformin-mediated Akt activation increased Akt substrate of 160 kDa (AS160) phosphorylation (0.51 ± 0.04 vs.1.03 ± 0.03,P =0.0041),promoted membrane translocation of glucose transporter 1,and increased glucose influx into the cells (4.78 ± 0.04 vs.5.47 ± 0.01,P ＜ 0.001).Contusion:This study suggested that targeting macrophage metabolism with new or existing drugs had therapeutic potential for the prevention and treatment of diabetes-accelerated atherosclerosis.

Reference

[1]Ye, Jinhui,Shi, Fuli,Lin, Hui,etc.Metformin Inhibits Chemokine Expression Through the AMPK/NF-kappa B Signaling Pathway[J].2018,38(9).
[2]Arai M,Uchiba M,Komura H,etc.Metformin, an antidiabetic agent, suppresses the production of tumor necrosis factor and tissue factor by inhibiting early growth response factor-1 expression in human monocytes in vitro.[J].2010,334(1).
[3]Griffin, Simon J.,Leaver, James K.,Irving, Greg J..Impact of metformin on cardiovascular disease: a meta-analysis of randomised trials among people with type 2 diabetes[J].2017,60(9).
[4]Biswas,S.K.,Chittezhath,M.,Shalova,I.N.,etc.Macrophage polarization and plasticity in health and disease[J].2012,53(1/3).
[5]M R, Munday.Regulation of mammalian acetyl-CoA carboxylase.[J].2002,30.
[6]Ismail-Beigi, Faramarz,Moghissi, Etie,Kosiborod, Mikhail,etc.Shifting Paradigms in the Medical Management of Type 2 Diabetes: Reflections on Recent Cardiovascular Outcome Trials[J].2017,32(9).
[7]Park JL,Zhn Q,Buller CL,etc.A GSK-3/TSC2/mTOR pathway regulates glucose uptake and GLUT1 glucose transporter expression[J].2008,295(3 Pt.1).
[8]Rena, Graham,Hardie, D. Grahame,Pearson, Ewan R..The mechanisms of action of metformin[J].2017,60(9).
[9]Zhou, Joseph,Massey, Scott,Story, Darren,etc.Metformin: An Old Drug with New Applications.[J].2018,19(10).
[10]Jha, Abhishek K.,Huang, Stanley Ching-Cheng,Sergushichev, Alexey,etc.Network Integration of Parallel Metabolic and Transcriptional Data Reveals Metabolic Modules that Regulate Macrophage Polarization[J].2015,42(3).
[11]Gerasimos,Siasos,Vasiliki,Tsigkou,Marinos,Kosmopoulos,etc.Mitochondria and cardiovascular diseases-from pathophysiology to treatment.[J].2018,6(12).
[12]Divya, Vats,Lata, Mukundan,Justin I, Odegaard,etc.Oxidative metabolism and PGC-1beta attenuate macrophage-mediated inflammation.[J].2006,4.
[13]Turrens JF.Mitochondrial formation of reactive oxygen species.[J].2003,552(Pt.2).
[14]Hansson GK,Nicoletti A,Caligiuri G,etc.Interleukin-10 deficiency increases atherosclerosis, thrombosis, and low-density lipoproteins in apolipoprotein E knockout mice.[J].2003,9(1).
[15]Joel N. Meyer,Tess C. Leuthner,Anthony L. Luz.Mitochondrial fusion, fission, and mitochondrial toxicity[J].2017,391.
[16]Wang, Ying,Tall, Alan R.,Wang, Nan,etc.Mitochondrial Oxidative Stress Promotes Atherosclerosis and Neutrophil Extracellular Traps in Aged Mice[J].2017,37(8).
[17]Virmani R,Burke AP,Kolodgie F.Morphological characteristics of coronary atherosclerosis in diabetes mellitus.[J].2006,22(Suppl.B).
[18]Bashir, Samina,Sharma, Yadhu,Elahi, Asif,etc.Macrophage polarization: the link between inflammation and related diseases[J].2016,65(1).
[19]Hayward, Rodney A.,Reaven, Peter D.,Wiitala, Wyndy L.,etc.Follow-up of Glycemic Control and Cardiovascular Outcomes in Type 2 Diabetes[J].2015,372(23).
[20]Yisang Yoon,Eugene W. Krueger,Barbara J. Oswald,etc.The Mitochondrial Protein hFis1 Regulates Mitochondrial Fission in Mammalian Cells through an Interaction with the Dynamin-Like Protein DLP1[J].2003,23(15).
[21]Staels B,Bories G,Zawadzki C,etc.Human Atherosclerotic Plaque Alternative Macrophages Display Low Cholesterol Handling but High Phagocytosis Because of Distinct Activities of the PPAR{gamma} and LXR{alpha} Pathways.[J].2011,108(8).
[22]K, Gottlob,N, Majewski,S, Kennedy,etc.Inhibition of early apoptotic events by Akt/PKB is dependent on the first committed step of glycolysis and mitochondrial hexokinase.[J].2001,15.
[23]Cipolat S,Martins de Brito O,Dal Zilio B,etc.OPA1 requires mitofusin 1 to promote mitochondrial fusion.[J].2004,101(45).
[24]Chiara, Buono,Carolyn E, Come,George, Stavrakis,etc.Influence of interferon-gamma on the extent and phenotype of diet-induced atherosclerosis in the LDLR-deficient mouse.[J].2003,23.
[25]Oh,J.,Riek,A.E.,Weng,S.,etc.Endoplasmic reticulum stress controls M2 macrophage differentiation and foam cell formation[J].2012,287(15).
[26]Anusha,Angajala,Sangbin,Lim,Joshua B,Phillips,etc.Diverse Roles of Mitochondria in Immune Responses: Novel Insights Into Immuno-Metabolism.[J].2018,9.
[27]Rémy, Boussageon,Theodora, Bejan-Angoulvant,Mitra, Saadatian-Elahi,etc.Effect of intensive glucose lowering treatment on all cause mortality, cardiovascular death, and microvascular events in type 2 diabetes: meta-analysis of randomised controlled trials.[J].2011,343.
[28]Han X,Kitamoto S,Wang H,etc.Interleukin-10 overexpression in macrophages suppresses atherosclerosis in hyperlipidemic mice.[J].2010,24(8).
[29]Battsetseg Batchuluun,Toyoshi Inoguchi,Noriyuki Sonoda,etc.Metformin and liraglutide ameliorate high glucose-induced oxidative stress via inhibition of PKC-NAD(P)H oxidase pathway in human aortic endothelial cells[J].2014,232(1).
[30]Yu, Emma P. K.,Reinhold, Johannes,Yu, Haixiang,etc.Mitochondrial Respiration Is Reduced in Atherosclerosis, Promoting Necrotic Core Formation and Reducing Relative Fibrous Cap Thickness[J].2017,37(12).
[31]Sarwar N,Gao P,Seshasai SR,etc.Diabetes mellitus, fasting blood glucose concentration, and risk of vascular disease: a collaborative meta-analysis of 102 prospective studies.[J].2010,375(9733).
[32]Cartee, Gregory D..Roles of TBC1D1 and TBC1D4 in insulin- and exercise-stimulated glucose transport of skeletal muscle[J].2015,58(1).
[33].Distribution of macrophage polarization markers in human atherosclerosis[J].2012,225(2).
[34]Freemerman,A.J.,Johnson,A.R.,Sacks,G.N.,etc.Metabolic reprogramming of macrophages: Glucose transporter 1 (GLUT1)-mediated glucose metabolism drives a proinflammatory phenotype[J].2014,289(11).
[35]Jordan P,Moniz S,Mendes AI,etc.Protein kinase WNK1 promotes cell surface expression of glucose transporter GLUT1 by regulating a Tre-2/USP6-BUB2-Cdc16 domain family member 4 (TBC1D4)-Rab8A complex.[J].2010,285(50).
[36]Feig, J.E.,Rong, J.X.,Shamir, R.,etc.HDL promotes rapid atherosclerosis regression in mice and alters inflammatory properties of plaque monocyte-derived cells[J].2011,108(17).
[37]Wang, Qilong,Zhang, Miao,Torres, Gloria,etc.Metformin Suppresses Diabetes-Accelerated Atherosclerosis via the Inhibition of Drp1-Mediated Mitochondrial Fission[J].2017,66(1).
[38]Tabas I,Bornfeldt KE.Insulin resistance, hyperglycemia, and atherosclerosis.[J].2011,14(5).
[39]Parathath S,Fisher EA,Feig JE,etc.Reversal of hyperlipidemia with a genetic switch favorably affects the content and inflammatory state of macrophages in atherosclerotic plaques.[J].2011,123(9).
[40]S C, Whitman,P, Ravisankar,H, Elam,etc.Exogenous interferon-gamma enhances atherosclerosis in apolipoprotein E-/- mice.[J].2000,157.