Apigenin Retards Atherogenesis by Promoting ABCA1-Mediated Cholesterol Efflux and Suppressing Inflammation

Background/Aims: The development of atherosclerosis is accompanied by escalating inflammation and lipid accumulation within blood vessel walls. ABCA1 plays a crucial role in mediating cholesterol efflux from macrophages, which protects against atherogenesis. This research was designed to explore the effects and underlying mechanisms of apigenin (4’, 5, 7-trihydroxyflavone) on ABCA1-mediated cellular cholesterol efflux and LPS-stimulated inflammation in RAW264.7 macrophages and apoE-/- mice. Methods: Expression of genes or proteins was examined by RT-PCR or western blot analysis. Liquid scintillation counting was used to detect percent cholesterol efflux. Cellular cholesterol content was measured using HPLC assay. The secretion levels of pro-inflammatory cytokines were quantified by ELISA assay. Atherosclerotic lesion sizes were determined with Oil Red O staining. The contents of macrophages and smooth muscle cells in atherosclerotic lesion were evaluated using immunohistochemistry. Plasma TC, TG, HDL-C and LDL-C levels in apoE-/- mice were evaluated using commercial test kits. Results: Apigenin potently increased ABCA1 expression through miR-33 repression in a dose- and time-dependent manner. Treatment with apigenin significantly increased ABCA1-mediated cholesterol efflux, and reduced TC, FC and CE levels in macrophage-derived foam cells. In LPS-treated macrophages, the expression levels of TLR-4, MyD88 and p-IκB-α as well as nuclear NF-κB p65 were decreased by the addition of apigenin. Moreover, apigenin markedly decreased secretion levels of several pro-inflammatory cytokines. Lastly, in LPS-challenged apoE-/- mice, apigenin administration augmented ABCA1 expression, decreased the contents of macrophages and smooth muscle cells in atherosclerotic lesion, reduced miR-33, TLR-4, and NF-κB p65 levels, improved plasma lipid profile and relieved inflammation, which results in less atherosclerotic lesion size. Conclusions: Taken together, these results suggest that apigenin may attenuate atherogenesis through up-regulating ABCA1-mediated cholesterol efflux and inhibiting inflammation.

[1]  A. Emmert,et al.  Apigenin Reduces NF-κB and Subsequent Cytokine Production as Protective Effect in a Rodent Animal Model of Lung Ischemia-Reperfusion Injury , 2018, Journal of investigative surgery : the official journal of the Academy of Surgical Research.

[2]  E. Georges,et al.  Apigenin-induced ABCC1-mediated efflux of glutathione from mature erythrocytes inhibits the proliferation of Plasmodium falciparum. , 2017, International journal of antimicrobial agents.

[3]  Meilin Xie,et al.  Apigenin protects against alcohol-induced liver injury in mice by regulating hepatic CYP2E1-mediated oxidative stress and PPARα-mediated lipogenic gene expression. , 2017, Chemico-biological interactions.

[4]  Peng Yang,et al.  Apigenin Restrains Colon Cancer Cell Proliferation via Targeted Blocking of Pyruvate Kinase M2-Dependent Glycolysis. , 2017, Journal of agricultural and food chemistry.

[5]  Min Zhang,et al.  Hsp27 promotes ABCA1 expression and cholesterol efflux through the PI3K/PKC&zgr;/Sp1 pathway in THP‐1 macrophages , 2017, European journal of pharmacology.

[6]  Yanwei Xing,et al.  Oxidative Stress-Mediated Atherosclerosis: Mechanisms and Therapies , 2017, Front. Physiol..

[7]  Yanan Zhu,et al.  Modafinil attenuates inflammation via inhibiting Akt/NF-κB pathway in apoE-deficient mouse model of atherosclerosis , 2017, Inflammopharmacology.

[8]  Fengxia Tu,et al.  Apigenin Ameliorates Post-Stroke Cognitive Deficits in Rats Through Histone Acetylation-Mediated Neurochemical Alterations , 2017, Medical science monitor : international medical journal of experimental and clinical research.

[9]  Hongxia Yang,et al.  Inhibition of TLR4 alleviates the inflammation and apoptosis of retinal ganglion cells in high glucose , 2017, Graefe's Archive for Clinical and Experimental Ophthalmology.

[10]  Y. Kawakami,et al.  Impact of TLR 2, TLR 4-activation on the Expression of ABCA1 and ABCG1 in Raw Cells. , 2017, Annals of clinical and laboratory science.

[11]  Fang Li,et al.  Apigenin Alleviates Endotoxin-Induced Myocardial Toxicity by Modulating Inflammation, Oxidative Stress, and Autophagy , 2017, Oxidative medicine and cellular longevity.

[12]  Pingping Shen,et al.  Apigenin, a modulator of PPAR&ggr;, attenuates HFD‐induced NAFLD by regulating hepatocyte lipid metabolism and oxidative stress via Nrf2 activation , 2017, Biochemical pharmacology.

[13]  Bin Huang,et al.  Tribbles homolog 1 enhances cholesterol efflux from oxidized low-density lipoprotein-loaded THP-1 macrophages. , 2017, Experimental and therapeutic medicine.

[14]  Weifeng Shen,et al.  DC-SIGN and Toll-like receptor 4 mediate oxidized low-density lipoprotein-induced inflammatory responses in macrophages , 2017, Scientific Reports.

[15]  Jiliang Wu,et al.  Apigenin Attenuates Adriamycin-Induced Cardiomyocyte Apoptosis via the PI3K/AKT/mTOR Pathway , 2017, Evidence-based complementary and alternative medicine : eCAM.

[16]  Jun Li,et al.  PTPRO Promotes Oxidized Low-Density Lipoprotein Induced Oxidative Stress and Cell Apoptosis through Toll-Like Receptor 4/Nuclear Factor κB Pathway , 2017, Cellular Physiology and Biochemistry.

[17]  T. Liu,et al.  Dihydromyricetin ameliorates atherosclerosis in LDL receptor deficient mice. , 2017, Atherosclerosis.

[18]  L. Joseph,et al.  Protective role of apigenin on rotenone induced rat model of Parkinson's disease: Suppression of neuroinflammation and oxidative stress mediated apoptosis. , 2017, Chemico-biological interactions.

[19]  Q. Pang,et al.  SIRT6 reduces macrophage foam cell formation by inducing autophagy and cholesterol efflux under ox‐LDL condition , 2017, The FEBS journal.

[20]  R. Benndorf,et al.  ABC Transport Proteins in Cardiovascular Disease—A Brief Summary , 2017, Molecules.

[21]  Yu-qi Fan,et al.  MicroRNA-9 Inhibits NLRP3 Inflammasome Activation in Human Atherosclerosis Inflammation Cell Models through the JAK1/STAT Signaling Pathway , 2017, Cellular Physiology and Biochemistry.

[22]  Dalin Li,et al.  Apigenin in the regulation of cholesterol metabolism and protection of blood vessels , 2017, Experimental and therapeutic medicine.

[23]  Yan Zhu,et al.  Hawthorn (Crataegus pinnatifida Bunge) leave flavonoids attenuate atherosclerosis development in apoE knock-out mice. , 2017, Journal of ethnopharmacology.

[24]  Xuelian Xu,et al.  Quercetin improves macrophage reverse cholesterol transport in apolipoprotein E-deficient mice fed a high-fat diet , 2017, Lipids in Health and Disease.

[25]  H. Chung,et al.  Role of Apigenin in Cancer Prevention via the Induction of Apoptosis and Autophagy , 2016, Journal of cancer prevention.

[26]  R. Liu,et al.  Pycnogenol Reduces Toll-Like Receptor 4 Signaling Pathway-Mediated Atherosclerosis Formation in Apolipoprotein E-Deficient Mice , 2016, Journal of cardiovascular pharmacology.

[27]  A. Näär,et al.  MicroRNA-33 Regulates the Innate Immune Response via ATP Binding Cassette Transporter-mediated Remodeling of Membrane Microdomains * , 2016, The Journal of Biological Chemistry.

[28]  Mukut Sharma,et al.  Cigarette Smoke Amplifies Inflammatory Response and Atherosclerosis Progression Through Activation of the H1R-TLR2/4-COX2 Axis , 2015, Front. Immunol..

[29]  C. Qiao,et al.  Pycnogenol attenuates atherosclerosis by regulating lipid metabolism through the TLR4–NF-κB pathway , 2015, Experimental & Molecular Medicine.

[30]  G. Liang,et al.  Apigenin protects blood-brain barrier and ameliorates early brain injury by inhibiting TLR4-mediated inflammatory pathway in subarachnoid hemorrhage rats. , 2015, International immunopharmacology.

[31]  K. Hirata,et al.  Anti-inflammatory and immune-modulatory therapies for preventing atherosclerotic cardiovascular disease. , 2015, Journal of cardiology.

[32]  Xiuqiong Fu,et al.  Inhibition of autophagy ameliorates atherogenic inflammation by augmenting apigenin-induced macrophage apoptosis. , 2015, International immunopharmacology.

[33]  B. Liu,et al.  Apigenin Attenuates Atherogenesis through Inducing Macrophage Apoptosis via Inhibition of AKT Ser473 Phosphorylation and Downregulation of Plasminogen Activator Inhibitor-2 , 2015, Oxidative medicine and cellular longevity.

[34]  Xi-Long Zheng,et al.  Chlamydia pneumoniae negatively regulates ABCA1 expression via TLR2-Nuclear factor-kappa B and miR-33 pathways in THP-1 macrophage-derived foam cells. , 2014, Atherosclerosis.

[35]  Hamid Nasri,et al.  Atherosclerosis: Process, Indicators, Risk Factors and New Hopes , 2014, International journal of preventive medicine.

[36]  T. Lehtimäki,et al.  MicroRNAs in the atherosclerotic plaque. , 2013, Clinical chemistry.

[37]  Min Zhang,et al.  Antagonism of Betulinic Acid on LPS-Mediated Inhibition of ABCA1 and Cholesterol Efflux through Inhibiting Nuclear Factor-kappaB Signaling Pathway and miR-33 Expression , 2013, PloS one.

[38]  Kai Yin,et al.  Foam cells in atherosclerosis. , 2013, Clinica chimica acta; international journal of clinical chemistry.

[39]  K. Moore,et al.  Macrophages in atherosclerosis: a dynamic balance , 2013, Nature Reviews Immunology.

[40]  C. Monaco,et al.  Toll-Like Receptors in Atherosclerosis , 2013, International journal of molecular sciences.

[41]  A. Papavassiliou,et al.  MicroRNAs: Novel diagnostic and prognostic biomarkers in atherosclerosis. , 2013, Current topics in medicinal chemistry.

[42]  A. Tall,et al.  Deficiency of ATP-Binding Cassette Transporters A1 and G1 in Macrophages Increases Inflammation and Accelerates Atherosclerosis in Mice , 2013, Circulation research.

[43]  Zhi-Sheng Jiang,et al.  Apolipoprotein A-I inhibits LPS-induced atherosclerosis in ApoE−/− mice possibly via activated STAT3-mediated upregulation of tristetraprolin , 2013, Acta Pharmacologica Sinica.

[44]  Chao-ke Tang,et al.  MicroRNA-467b targets LPL gene in RAW 264.7 macrophages and attenuates lipid accumulation and proinflammatory cytokine secretion. , 2012, Biochimie.

[45]  N. Mohan,et al.  Combination of N-(4-hydroxyphenyl) retinamide and apigenin suppressed starvation-induced autophagy and promoted apoptosis in malignant neuroblastoma cells , 2011, Neuroscience Letters.

[46]  Xiang-Ling Tan,et al.  Optimization of ultrasonic-assisted enzymatic hydrolysis for the extraction of luteolin and apigenin from celery. , 2011, Journal of food science.

[47]  X. Deng,et al.  Tristetraprolin-dependent Post-transcriptional Regulation of Inflammatory Cytokine mRNA Expression by Apolipoprotein A-I , 2011, The Journal of Biological Chemistry.

[48]  T. Nakayama,et al.  Inflammation, a Link between Obesity and Cardiovascular Disease , 2010, Mediators of inflammation.

[49]  K. Moore,et al.  MiR-33 Contributes to the Regulation of Cholesterol Homeostasis , 2010, Science.

[50]  D. Scott‐Algara,et al.  Antiinflammatory and antiatherogenic effects of the NF-kappaB inhibitor acetyl-11-keto-beta-boswellic acid in LPS-challenged ApoE-/- mice. , 2008, Arteriosclerosis, thrombosis, and vascular biology.

[51]  D. Scott‐Algara,et al.  Antiinflammatory and Antiatherogenic Effects of the NF-&kgr;B Inhibitor Acetyl-11-Keto-β-Boswellic Acid in LPS-Challenged ApoE−/− Mice , 2007 .

[52]  J. Kang,et al.  Attenuation of monocyte adhesion and oxidised LDL uptake in luteolin-treated human endothelial cells exposed to oxidised LDL , 2007, British Journal of Nutrition.

[53]  G. Stoll,et al.  Inflammation and Atherosclerosis: Novel Insights Into Plaque Formation and Destabilization , 2006, Stroke.

[54]  A. Remaley,et al.  Lipopolysaccharide Down Regulates Both Scavenger Receptor B1 and ATP Binding Cassette Transporter A1 in RAW Cells , 2002, Infection and Immunity.

[55]  M. Fishbein,et al.  Toll-Like Receptor-4 Is Expressed by Macrophages in Murine and Human Lipid-Rich Atherosclerotic Plaques and Upregulated by Oxidized LDL , 2001, Circulation.

[56]  G. Liang,et al.  MD2 mediates angiotensin II-induced cardiac inflammation and remodeling via directly binding to Ang II and activating TLR4/NF-κB signaling pathway , 2016, Basic Research in Cardiology.

[57]  Junfei Jin,et al.  TLR4 antagonist reduces early-stage atherosclerosis in diabetic apolipoprotein E-deficient mice. , 2013, The Journal of endocrinology.

[58]  Zhi-Sheng Jiang,et al.  Interleukin-18 and interleukin-12 together downregulate ATP-binding cassette transporter A1 expression through the interleukin-18R/nuclear factor-κB signaling pathway in THP-1 macrophage-derived foam cells. , 2012, Circulation journal : official journal of the Japanese Circulation Society.

[59]  Kwan-Kyu Park,et al.  Melittin inhibits atherosclerosis in LPS/high-fat treated mice through atheroprotective actions. , 2011, Journal of atherosclerosis and thrombosis.

[60]  K. Watanabe,et al.  Enhancement of phenylephrine-induced contraction in the isolated rat aorta with endothelium by H2O-extract from an Oriental medicinal plant Leonuri herba. , 2001, Japanese journal of pharmacology.

[61]  C. Shin,et al.  Annals of the New York Academy of Sciences Microrna–target Interactions: New Insights from Genome-wide Approaches , 2022 .