Glycated ferritin increases the in vitro expression of TLR2 and TLR4 in peripheral blood CD14+ cells obtained from patients with prediabetes
Keywords:serum ferritin, glycated ferritin, prediabetes, toll-like receptors
- Glycated ferritin increases the expression of Toll-like receptors (TLR2, TLR4) in CD14+ blood leucocytes derived from healthy subjects. It is not known if a similar response occurs in subjects with prediabetes, a low-grade chronic inflammatory condition.
- We compared the expression of both receptors in PBMC CD14+ cells from prediabetic and normoglycemic subjects.
- Glycated ferritin increased TLR2, TLR4, IL-6 and IL-8 expression in cells from prediabetic subjects more than that from normoglycemic subjects. An increase in basal TLR4 expression was only found in cells from prediabetic subjects. This could be part of low-grade inflammation, which could be increased by glycated ferritin.
Abstract: Serum ferritin is a widely-used marker of inflammation in prediabetes, diabetes and atherosclerosis. In these cases, progressive endothelial damage may involve the participation of toll-like receptors (TLR). The aim of this study was to determine the expression of TLR2 and TLR4 in peripheral blood mononuclear cell (PBMC)-derived CD14+ cells from subjects with prediabetes and with a high level of serum ferritin both at baseline and after in vitro cell stimulation with glycated ferritin. Blood samples were drawn from 22 subjects (13 with prediabetes and 9 with normoglycemia). Serum ferritin levels were measured by ELISA, while the expression of TLR2 and TLR4 in PBMC-derived CD14+ cells was determined by flow cytometry. IL-6 and IL-8 cytokines in PBMC-derived CD14+ supernatants were measured by ELISA. Subjects with prediabetes had a higher baseline expression of TLR4 in PBMC-derived CD14+ cells than was observed in cells from normoglycemic subjects (p<0.05). Glycated ferritin increased the expression of both TLR2 and TLR4 as well as IL-6 and IL-8 in PBMC-derived CD14+ cells from subjects with prediabetes when compared to normoglycemic subjects (p<0.05). We concluded that in prediabetes, the increased basal expression of TLR4 could be part of the low-grade inflammation, which could be increased by glycated ferritin.
Received: March 31, 2020; Revised: May 6, 2020; Accepted: May 23, 2020; Published online: May 28, 2020
How to cite this article: Galván-Moroyoqui JM, Martínez-Soto JM, López-Soto LF, Soto-Guzmán JA, Camacho-Villa AY, Alvarez-Hernandez G, Mata-Pineda AL, Candia-PlataMC. Glycated ferritin increases the in vitro expression of TLR2 and TLR4 in peripheral blood CD14+ cells obtained from patients with prediabetes. Arch Biol Sci. 2020:72(3):305-12.
Huang Z, Chen C, Li S, Chen C, Li S, Kong F, Shan P, Huang W. Serum markers of endothelial dysfunction and inflammation increase in hypertension with prediabetes mellitus. Genet Test Mol Biomarkers. 2016;20(6):322-7.
Meng G, Yang H, Bao X, Zhang Q, Liu L, Wu H, Du H, Xia Y, Shi H, Guo X, Liu X, Li C, Su Q, Gu Y, Fang L, Yu F, Sun S, Wang X, Zhou M, Jia Q, Guo Q, Song K, Huang G, Wang G, Wu Y, Niu K. Increased serum ferritin levels are independently related to incidence of prediabetes in adult populations. Diabetes Metab. 2017;43(2):146-53.
Kato K, Otsuka T, Saiki Y, Kobayashi N, Nakamura T, Kon Y, Kawada T. Association between elevated C-reactive protein levels and prediabetes in adults, particularly impaired glucose tolerance. Can J Diabetes. 2019;43(1):40-5.
Sharifi F, Nasab NM, Zadeh HJ. Elevated serum ferritin concentrations in prediabetic subjects. Diab Vasc Dis Res. 2008;5(1):15-8.
Aregbesola A, Virtanen JK, Voutilainen S, Mursu J, Lagundoye A, Kauhanen J, Toumainen TP. Serum ferritin and glucose homeostasis: change in the association by glycaemic state. Diabetes Metab Res Rev. 2015;31(5):507-14.
Zhou Y, Liu T, Tian C, Kang P, Jia C. Association of serum ferritin with coronary artery disease. Clin Biochem. 2012;45(16-17):1336-41.
Kell DB, Pretorius E. Serum ferritin is an important inflammatory disease marker, as it is mainly a leakage product from damaged cells. Metallomics. 2014;6:748-73.
Curtiss LK, Tobias PS. Emerging role of Toll-like receptors in atherosclerosis. J Lipid Res. 2009;50:S340-5.
Hodgkinson CP, Ye S. Toll-Like receptors, their ligands, and atherosclerosis. Sci World J. 2011;11:437-53.
Tapping RI, Tobias PS. Mycobacterial lipoarabinomannan mediates physical interactions between TLR1 and TLR2 to induce signaling. J Endotoxin Res. 2003;9(4):264-8.
Roeder A, Kirschning CJ, Rupec RA, Schaller M, Weindl G, Korting HC. Toll-like receptors as key mediators in innate antifungal immunity. Med Mycol. 2004;42(6):485-98.
Massari P, Visintin A, Gunawardana J, Halmen KA, King CA, Golenbock DT, Wetzler LM. Meningococcal porin PorB binds to TLR2 and requires TLR1 for signaling. J Immunol. 2006;176(4):2373-80.
Wong FS, Wen L. Toll-like receptors and diabetes. Ann N Y Acad Sci. 2008;1150:123-132.
Ionita MG, Arslan F, de Kleijn DP, Pasterkamp G. Endogenous inflammatory molecules engage Toll-like receptors in cardiovascular disease. J Innate Immun. 2010;2(4):307-15.
Katakami N. Mechanism of development of atherosclerosis and cardiovascular disease in diabetes mellitus. J Atheroscler Thromb. 2018;25(1):27‐39.
Dasu MR, Devaraj S, Zhao L, Devaraj S, Zhao L, Hwang DH, Jialal I. High glucose induces Toll-like receptor expression in human monocytes mechanism of activation. Diabetes. 2008;57:3090-8.
Jialal I, Kaur H. The role of Toll-Like receptors in diabetes-induced inflammation: Implications for vascular complications. Curr Diab Rep. 2012;12(2):172-9.
Cheng A, Dong YY, Zhu FX, Liu Y, Hou FF, Nie J. AGE-LDL activates Toll like receptor 4 pathway and promotes inflammatory cytokines production in renal tubular epithelial cells. Int J B Sci. 2013;9:94-107.
Galván-Moroyoqui JM, Candia-Plata MC, Martínez-Soto JM, Soto-Guzmán JA, Camacho-Villa AY, López-Soto LF. Glycated ferritin induces activation and expression of tlr2 and tlr4 in human peripheral blood macrophages. The Pharma Innovation Journal. 2015;3(12):44-8.
López-Soto LF G-MJ, Galvan-Moroyoqui JM, Martínez-Soto MJ, Almada BM, Rosales RA, Álvarez-Hernández G, Camacho-Villa A, Bolado-Martínez E, Soto-Guzmán JA, Candia-Plata MC. Glycoxidated ferritin induces the release of microparticles positive for toll-like receptors derived from peripheral blood CD14+ cells. Arch Biol Sci. 2017;69(3):383-90.
American Diabetes Association. Classification and diagnosis of diabetes: Standards of Medical Care in Diabetes 2018. Diabetes Care. 2018;41(Suppl 1):S13-27.
Sheu WH, Chen YT, Lee WJ, Wang CW, Lin LY. A relationship between serum ferritin and the insulin syndrome is present in non-diabetic women but not in non-diabetic men. Clin Endocrinol. 2003;58(3):380-5.
Kim CH, Kim HK, Bae SJ, Park JY, Lee KU. Association of elevated serum ferritin concentration with insulin resistance and impaired glucose metabolism in Korean men and women. Metabolism. 2011;60(3):414-20.
Dekker LH, Nicolaou M, van der A DL, Dekker LH, Nicolaou M, van der A DL, Busschers WB, Brewster LM, Snijder MB, Stronks K and van Valkengoed IG. Sex differences in the association between serum ferritin and fasting glucose in type 2 diabetes among South Asian Surinamese, African Surinamese, and ethnic Dutch: the population-based SUNSET study. Diabetes Care. 2013;36(4):965-71.
Batchuluun B, Matsumata T, Batchuluun B, Erdenebileg N, Tsagaantsooj G, Boldbaatar K, Khasag A. Serum ferritin level is higher in poorly controlled patients with type 2 diabetes and people without diabetes, aged over 55 years. Diabet Med. 2014;31(4):419-24.
Alam F, Memor AS, Fatima SS. Increased body mass index may lead to hyperferritinemia irrespective of body iron stores. Pak J Med Sci. 2015;31(6):1521-6.
Friedewald WT, Levy RI, Fredrickson DS. Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge. Clin Chem. 1972;18(6):499-502.
Matthews DR, Hosker JP, Rudenski AS, Naylor BA, Treacher DF, Turner RC. Homeostasis model assessment-insulin resistance and beta-cell function from fasting plasma-glucose and insulin concentrations in man. Diabetologia. 1985;28(7):412-9.
Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248-54.
Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227:680-5.
Lichtenauer M, Nickl S, Hoetzenecker K, Mangold A, Moser B, Zimmermann M, Hacker S, Niederpold T, Mitterbauer A, Ankersmit H. Phosphate buffered saline containing calcium and magnesium elicits increased secretion of Interleukin-1 Receptor Antagonist. Lab Medicine. 2009.40;5(5):290-3.
Kreuser J, Bach N, Forler D, Sieber S. Target discovery of acivicin in cancer cells elucidates its mechanism of growth inhibition. Electronic supplementary information (ESI) available: Synthesis, cloning, protein expression, purification and biochemical assays. Chem Sci. 2014;6(1):237-45.
Rose L, Kadayakkara D, Wang G, Bar-Shir A, Helfer B, O'Hanlon C, Kraitchman D, Rodriguez R, Bulte J. Fluorine-19 labeling of stromal vascular fraction cells for clinical imaging applications. Stem Cells Transl Med. 2015;4(12):1472-81.
Ikeda Y, Adachi Y, Ishibashi K, Miura N, Ohno N. Activation of Toll-Like receptor-mediated NF-κB by zymosan-derived water-soluble fraction: Possible contribution of endotoxin-like substances. Immunopharmacol Immunotoxicol. 2005;27(2):285-98.
Taghavi M, Mortaz E, Khosravi A, Vahedi G, Folkerts G, Varahram M, Kazempour-Dizaji M, Garssen J, Adcock IM. Zymosan attenuates melanoma growth progression, increases splenocyte proliferation and induces TLR-2/4 and TNF-α expression in mice. J Inflamm (Lond). 2018;15:5.
Wang W, Knovich MA, Coffman LG, Torti FM, Torti SV. Serum ferritin: Past, present and future. Biochim Biophys Acta. 2010;1800(8):760-9.
Hodgkinson CP, Laxton RC, Patel K,Ye S. Advanced glycation end-product of low density lipoprotein activates the Toll-like 4 receptor pathway implications for diabetic atherosclerosis. Arterioscler Thromb Vasc Biol. 2008;28:2275-2281
Choi KH, Park JW, Kim HY, Choi KH, Park JW, Kim HY, Kim YH, Kim SM, Son YH, Park YC, Eo SK, Kim K. Cellular factors involved in CXCL8 expression induced by glycated serum albumin in vascular smooth muscle cells. Atherosclerosis. 2010;209:58-65
Cragg SJ, Wagstaff M, Worwood M. Detection of a glycosylated subunit in human serum ferritin. Biochem J. 1981;199:565-71.
Kernan KF, Carcillo JA. Hyperferritinemia and inflammation. Int Immunol. 2017;29:401-9.
Dasu MR, Devaraj S, Park S, Devaraj S, Park S, Jialal I. Increased Toll-like receptor (TLR) activation and TLR ligands in recently diagnosed type 2 diabetic subjects. Diabetes Care. 2010;33:861-8.
Huang Y, Guo W, Zeng J, Chen G, Sun W, Zhang X, Tian W. Prediabetes enhances periodontal inflammation consistent with activation of Toll-like receptor-mediated nuclear factor-kappa B pathway in rats. J Periodontol. 2016;87:E64-74.
De Loera-Rodriguez CO, Delgado-Rizo V, Alvarado-Navarro A, De Loera-Rodriguez CO, Delgado-Rizo V, Alvarado-Navarro A, Agraz-Cibrian JM, Segura-Ortega JE, Fafutis-Morris M. Over-expression of TLR4-CD14, pro-inflammatory cytokines, metabolic markers and NEFAs in obese non-diabetic Mexicans. J Inflamm. 2014;11:39.
Kaur H, Chien A, Jialal I. Hyperglycemia induces Toll like receptor 4 expression and activity in mouse mesangial cells: relevance to diabetic nephropathy. A J Renal Physiol. 2012;303:F1145-50.
Cimini FA, Barchetta I, Porzia A, Mainiero F, Costantino C, Bertoccini L, Ceccarelli V, Morini S, Baroni MG, Lenzi A, Cavallo MG. Circulating IL-8 levels are increased in patients with type 2 diabetes and associated with worse inflammatory and cardiometabolic profile. Acta Dabetol. 2017;54(10):961-7.