Evaluation of Toll-Like Receptor 2 and 4 RNA Expression and the Cytokine Profile in Postmenopausal Women with Metabolic Syndrome

Objective To evaluate the gene expression of Toll-Like (TLR-2 and TLR-4) receptors and cytokine profile in postmenopausal women with or without metabolic syndrome (MetS). Methods In this cross-sectional study, 311 Brazilian women (age≥45 years and amenorrhea≥12 months) were included. Women showing three or more of the following diagnostic criteria were diagnosed as positive for MetS: waist circumference>88 cm, triglycerides≥150 mg/dL, HDL cholesterol<50 mg/dL, blood pressure≥130/85 mmHg, and fasting glucose≥100 mg/dL. The expression of TLR-2 and TLR-4 in peripheral blood was evaluated by RNA extraction and subsequent real time PCR analysis. The cytokine profile, tumor necrosis factor alpha (TNF-α) and interleukins 1β, 6, and 10, were measured by ELISA. Results The expression of TLR-2 RNA was demonstrated in 32.5% and TLR-4 in 20.6% of the subjects. There was no association between the expression of TLR-2 and TLR-4 and the presence or absence of MetS (P>0.05). A greater production of IL-6 was associated with TLR-2 and TLR-4 expressions and greater production of TNF-α was associated only with TLR-2 expression (P>0.05). Only the lower quartile of IL-10 was associated with the presence of the MetS (P>0.05). Conclusions TLR-2 and TLR-4 expressions were associated with increased pro-inflammatory cytokines, IL-6 and TNF-α, with no association with biomarkers of MetS. The low concentrations of IL-10 may suggest an anti-inflammatory modulation in postmenopausal women with MetS.

[1]  J. Mckenney,et al.  Executive Summary of The Third Report of The National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, And Treatment of High Blood Cholesterol In Adults (Adult Treatment Panel III). , 2001, JAMA.

[2]  C. Monaco,et al.  The Expression and Functions of Toll-Like Receptors in Atherosclerosis , 2010, Mediators of inflammation.

[3]  S. Yusuf,et al.  Effect of potentially modifiable risk factors associated with myocardial infarction in 52 countries (the INTERHEART study): case-control study , 2004, The Lancet.

[4]  H. Uemura,et al.  Changes in serum cytokine concentrations during the menopausal transition. , 2007, Maturitas.

[5]  L. Joosten,et al.  Toll-like receptor 2 controls expansion and function of regulatory T cells. , 2006, The Journal of clinical investigation.

[6]  François Mach,et al.  Inflammation and Atherosclerosis , 2004, Herz.

[7]  N. Unwin,et al.  Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III) Detection, Evaluation, and Treatment of High Blood Cholesterol Education Program (NCEP) Expert Panel on Executive Summary of the Third Report of the National , 2009 .

[8]  A. Aljada,et al.  Contemporary Reviews in Cardiovascular Medicine Metabolic Syndrome A Comprehensive Perspective Based on Interactions Between Obesity, Diabetes, and Inflammation , 2022 .

[9]  M. Netea,et al.  Toll‐like receptor‐4 Asp299Gly polymorphism does not influence progression of atherosclerosis in patients with familial hypercholesterolaemia , 2004, European journal of clinical investigation.

[10]  I. Kubota,et al.  Central role of endogenous Toll-like receptor-2 activation in regulating inflammation, reactive oxygen species production, and subsequent neointimal formation after vascular injury. , 2006, Biochemical and biophysical research communications.

[11]  C. Ferrario,et al.  Role of the renin-angiotensin-aldosterone system and proinflammatory mediators in cardiovascular disease. , 2006, The American journal of cardiology.

[12]  G. Krikun,et al.  Lipopolysaccharide Appears to Activate Human Endometrial Endothelial Cells Through TLR‐4‐Dependent and TLR‐4‐Independent Mechanisms , 2012, American journal of reproductive immunology.

[13]  C. Franceschi,et al.  Genetic polymorphisms of inflammatory cytokines and myocardial infarction in the elderly , 2006, Mechanisms of Ageing and Development.

[14]  B. Franklin,et al.  American College of Sports Medicine position stand. Quantity and quality of exercise for developing and maintaining cardiorespiratory, musculoskeletal, and neuromotor fitness in apparently healthy adults: guidance for prescribing exercise. , 2011, Medicine and science in sports and exercise.

[15]  J. Gussekloo,et al.  The Leiden 85-plus Study , 2000 .

[16]  Claude Lenfant,et al.  Definition of Metabolic Syndrome: Report of the National Heart, Lung, and Blood Institute/American Heart Association Conference on Scientific Issues Related to Definition , 2004, Arteriosclerosis, thrombosis, and vascular biology.

[17]  K. Reynolds,et al.  The obese without cardiometabolic risk factor clustering and the normal weight with cardiometabolic risk factor clustering prevalence and correlates of 2 phenotypes among the US population (NHANES 1999-2004) , 2008 .

[18]  K. Reynolds,et al.  The obese without cardiometabolic risk factor clustering and the normal weight with cardiometabolic risk factor clustering: prevalence and correlates of 2 phenotypes among the US population (NHANES 1999-2004). , 2008, Archives of internal medicine.

[19]  K. Esposito,et al.  Cytokine pattern in postmenopause. , 2002, Maturitas.

[20]  G. Hansson,et al.  Expression of Toll-Like Receptors in Human Atherosclerotic Lesions: A Possible Pathway for Plaque Activation , 2002, Circulation.

[21]  S. Anker,et al.  A frequent toll-like receptor (TLR)-2 polymorphism is a risk factor for coronary restenosis , 2005, Journal of Molecular Medicine.

[22]  S Akira,et al.  Toll receptor families: structure and function. , 2004, Seminars in immunology.

[23]  M. Netea,et al.  Toll-like receptors on regulatory T cells: expanding immune regulation. , 2006, Trends in immunology.

[24]  J. Carvalheira,et al.  Inhibition of toll-like receptor 2 expression improves insulin sensitivity and signaling in muscle and white adipose tissue of mice fed a high-fat diet. , 2008, The Journal of endocrinology.

[25]  S. Akira,et al.  Toll-like receptors and innate immunity , 2006, Journal of Molecular Medicine.

[26]  B. Geloneze,et al.  The threshold value for insulin resistance (HOMA-IR) in an admixtured population IR in the Brazilian Metabolic Syndrome Study. , 2006, Diabetes research and clinical practice.

[27]  A. Pfeufer,et al.  Toll-like receptor 4 gene polymorphisms and myocardial infarction: no association in a Caucasian population. , 2006, European heart journal.

[28]  P. Tobias,et al.  TLR2 in murine atherosclerosis , 2008, Seminars in Immunopathology.

[29]  R. Marfella,et al.  Association of low interleukin-10 levels with the metabolic syndrome in obese women. , 2003, The Journal of clinical endocrinology and metabolism.

[30]  Yup Kang,et al.  IL-6 induction of TLR-4 gene expression via STAT3 has an effect on insulin resistance in human skeletal muscle , 2013, Acta Diabetologica.

[31]  A. D. de Craen,et al.  Low production capacity of interleukin-10 associates with the metabolic syndrome and type 2 diabetes : the Leiden 85-Plus Study. , 2002, Diabetes.

[32]  Chiang-Ching Huang,et al.  Activated TLR Signaling in Atherosclerosis among Women with Lower Framingham Risk Score: The Multi-Ethnic Study of Atherosclerosis , 2011, PloS one.

[33]  P. Verpillat,et al.  Association of the Toll-Like Receptor 4 Gene Asp299Gly Polymorphism With Acute Coronary Events , 2003, Arteriosclerosis, thrombosis, and vascular biology.

[34]  F. Ohsuzu,et al.  Characterization of the expression of TLR2 (toll-like receptor 2) and TLR4 on circulating monocytes in coronary artery disease. , 2005, Journal of atherosclerosis and thrombosis.

[35]  S. Ye,et al.  TLR4 Asp299Gly polymorphism is not associated with coronary artery stenosis. , 2003, Atherosclerosis.

[36]  Martin Bendszus,et al.  Inflammation and Atherosclerosis: Novel Insights Into Plaque Formation and Destabilization , 2006, Stroke.

[37]  Shizuo Akira,et al.  TLR signaling pathways. , 2004, Seminars in immunology.

[38]  L. Kuller,et al.  Epidemiology of cytokines: the Women On the Move through Activity and Nutrition (WOMAN) Study. , 2008, American journal of epidemiology.

[39]  R. Dias,et al.  Metabolic syndrome and its associated risk factors in Brazilian postmenopausal women , 2009, Climacteric : the journal of the International Menopause Society.

[40]  Frank B. Hu,et al.  Abdominal Obesity and the Risk of All-Cause, Cardiovascular, and Cancer Mortality: Sixteen Years of Follow-Up in US Women , 2008, Circulation.

[41]  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.

[42]  A. Boyle,et al.  The novel role of C-reactive protein in cardiovascular disease: risk marker or pathogen. , 2006, International journal of cardiology.

[43]  J. Kaski,et al.  Serum Levels of the Antiinflammatory Cytokine Interleukin-10 Are Decreased in Patients With Unstable Angina , 2001, Circulation.

[44]  P. Chedraui,et al.  Pro-inflammatory cytokine levels in postmenopausal women with the metabolic syndrome , 2011, Gynecological endocrinology : the official journal of the International Society of Gynecological Endocrinology.

[45]  J. C. Kaski,et al.  Interleucina-10 y enfermedad coronaria , 2002 .

[46]  R. Kimmig,et al.  TLR3 and TLR4 expression in healthy and diseased human endometrium , 2008, Reproductive biology and endocrinology : RB&E.

[47]  S. Crawford,et al.  Menopause and the metabolic syndrome: the Study of Women's Health Across the Nation. , 2008, Archives of internal medicine.

[48]  H. Schild,et al.  Heat shock proteins as ligands of toll-like receptors. , 2002, Current topics in microbiology and immunology.

[49]  G. Rosano,et al.  The Metabolic Syndrome in Women , 2006, Women's health.

[50]  S. Akira,et al.  Toll-like receptors: critical proteins linking innate and acquired immunity , 2001, Nature Immunology.

[51]  J. Carvalheira,et al.  Loss-of-Function Mutation in Toll-Like Receptor 4 Prevents Diet-Induced Obesity and Insulin Resistance , 2007, Diabetes.

[52]  J. Kaski,et al.  [Interleukin-10 and coronary disease]. , 2002, Revista espanola de cardiologia.

[53]  P. Libby.  Inflammation in atherosclerosis , 2002, Nature.

[54]  S. Devaraj,et al.  Increased Toll-Like Receptor Activity in Patients With Metabolic Syndrome , 2012, Diabetes Care.

[55]  A. Aljada,et al.  Circulating Mononuclear Cells in the Obese Are in a Proinflammatory State , 2004, Circulation.

[56]  G. Ertl,et al.  Mechanisms of Disease: Toll-like receptors in cardiovascular disease , 2007, Nature Clinical Practice Cardiovascular Medicine.

[57]  J. Manson,et al.  Circulating inflammatory and endothelial markers and risk of hypertension in white and black postmenopausal women. , 2011, Clinical chemistry.

[58]  P. Froguel,et al.  Inflammatory Role of Toll-Like Receptors in Human and Murine Adipose Tissue , 2010, Mediators of inflammation.

[59]  A. Parker,et al.  Modulation of Toll-Like Receptor Signalling as a New Therapeutic Principle , 2011, Mediators of inflammation.

[60]  F. Benencia,et al.  Toll-Like Receptors as Novel Therapeutic Targets for Ovarian Cancer , 2012, ISRN oncology.

[61]  J. Suttles,et al.  The Role of JAK-3 in Regulating TLR-Mediated Inflammatory Cytokine Production in Innate Immune Cells , 2013, The Journal of Immunology.

[62]  H. Methe,et al.  Expansion of Circulating Toll-Like Receptor 4–Positive Monocytes in Patients With Acute Coronary Syndrome , 2005, Circulation.

[63]  S. Shaw,et al.  The Emerging Role of TLR and Innate Immunity in Cardiovascular Disease , 2012, Cardiology research and practice.

[64]  J. Eckel,et al.  Adipose tissue inflammation: novel insight into the role of macrophages and lymphocytes , 2010, Current opinion in clinical nutrition and metabolic care.

[65]  P. Ridker,et al.  C-reactive protein, inflammation, and coronary risk. , 2000, The Medical clinics of North America.

[66]  A. Tedgui,et al.  Anti-Inflammatory Mechanisms in the Vascular Wall , 2001, Circulation research.