Expression of the receptor of advanced glycation end-products (RAGE) and membranal location in peripheral blood mononuclear cells (PBMC) in obesity and insulin resistance

Document Type: Original Article

Authors

1 Instituto de Investigación en Enfermedades Crónico-Degenerativas, Departamento de Biología Molecular y Genómica, C.U.C.S, Universidad de Guadalajara, Guadalajara, Jalisco, México

2 Programa de Doctorado en Ciencias en Biología Molecular en Medicina. Departamento de Biología Molecular y Genómica, C.U.C.S, Universidad de Guadalajara, Guadalajara, Jalisco, México

3 Laboratorio de Investigación en Microbiología, Departamento de Microbiología y Patología, C.U.C.S, Universidad de Guadalajara, Guadalajara, Jalisco, México

Abstract

Objective(s): The present study aimed to evaluate the receptor of advanced glycation end-products (RAGE), NF-kB, NRF2 gene expression, and RAGE cell distribution in peripheral blood mononuclear cells (PBMC) in subjects with obesity and IR compared with healthy subjects.
Materials and Methods: The mRNA expression levels of RAGE, NF-kB, NRF2, and GAPDH were determined in PBMC by qPCR in 20 obese (OB), 17 obese with insulin resistance (OB-IR) subjects, and 20 age and sex-matched healthy subjects (HS). RAGE protein expression and its localization were determined by Western Blot and immunocytochemistry (ICC) analysis, total soluble RAGE (sRAGE) and MCP-1 plasma levels by ELISA.
Results: RAGE, NF-kB, and NRF2 genes mRNA expression in PBMCs did not show variation between groups. RAGE protein was lower in OB and OB-IR groups; RAGE was located predominantly on the cell-surface in the OB-IR group compared to the HS group (22% vs 9.5%, P<0.001). OB-IR group showed lower sRAGE plasma levels, and correlated negatively with HOMA-IR, ALT parameters (r= -0.374, r= -0.429, respectively), and positively with NFE2L2 mRNA (r= 0.540) PConclusion: In this study, OB-IR subjects did not reflect significant differences in gene expression; however, correlations detected between sRAGE, biochemical parameters, and NRF2, besides the predominant RAGE distribution on the cell membrane in PBMC could be evidence of the early phase of the inflammatory cascade and the subsequent damage in specific tissues in subjects with OB-IR.

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Main Subjects


1. World Health Organization (WHO). Obesity: preventing and managing the global epidemic. Available online: http://www.who.int/topics/obesity/en/  (accessed on December 2018).

2.Hernández M, Rivera J, Shamah T, Cuevas L, Gómez L, Gaona E, García D, et al. Encuesta Nacional de Salud y Nutrición de Medio Camino (ENSANUT MC 2016). Available online: http://ensanut.insp.mx/ensanut2016/descarga_bases.php#.WoyFH-ejnIU (accessed  on December 2018). (In Spanish).

3.Zaki M, Kamal S, Kholousi S, El-Bassyouni HT, Yousef W, Reyad H, et al. Serum soluble receptor of advanced glycation end products and risk of metabolic syndrome in egyptian obese women. EXCLI J 2017; 16:973-980.

4.De Mello VDF, Kolehmainen M, Pulkkinen L, Schwab U, Mager U, Laaksonen DE, et al. Downregulation of genes involved in NFκB activation in peripheral blood mononuclear cells after weight loss is associated with the improvement of insulin sensitivity in individuals with the metabolic syndrome: The GENOBIN study. Diabetologia 2008; 51:2060-2067.

5.Schmidt AM. 2016 ATVB plenary lecture: receptor for advanced glycation endproducts and implications for the pathogenesis and treatment of cardiometabolic disorders: spotlight on the macrophage. Arterioscler Thromb Vasc Biol 2017; 37:613-621.

6.National Center for Biotechnology Information (NCBI). Available online: https://www.ncbi.nlm.nih.gov/nuccore/?LinkName=gene_nuccore_refseqrna&from_uid=177 (accessed on December 2018).

7.Drinda S, Franke T, Eidner C, Schmidt C, Rüster T, Bondeva G, et al. Decreased RAGE expression in peripheral blood mononuclear cells of patients with rheumatoid arthritis. Clin Exp Rheumatol 2009; 27:483-490.

8.Abdelsalam RM, Safar MM. Neuroprotective effects of vildagliptin in rat rotenone Parkinson’s disease model: role of RAGE-NFkB and Nrf2-antioxidant signaling pathways. J Neurochem 2015; 133:700-707.  

9.World Medical Association. World Medical Association Declaration of Helsinki: ethical principles for medical research involving human subjects. JAMA 2013; 310:2191–2194.

10.Matthews DR, Hosker JR, 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; 28:412-419.

11.Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 2001; 29:45e.

12.Su XD, Li SS, Tian YQ, Zhang ZY, Zhang GZ, Wang LX. Elevated serum levels of advanced glycation end products and their monocyte receptors in patients with type 2 diabetes. Arch Med Res 2011; 42:596-601.

13.Czec MP. Insulin action and resistance in obesity and type 2 diabetes. Nat Med 2017; 23:804-814.

14.Gentile AM, Lhamyani S, Coín-Aragüez L, Oliva-Olivera W, Zayed H, Vega-Rioja A, et al. RPL13A and EEF1A1 are suitable reference genes for qPCR during adipocyte differentiation of vascular stromal cells from patients with different BMI and HOMA-IR. PLoS One 2016; 11:e0157002.

15.Biswas SK, Mohtarin S, Mudi SR, Anwar T, Banu LA, Alam SM, et al. Relationship of soluble RAGE with insulin resistance and beta cell function during development of type 2 diabetes mellitus. J Diabetes Res 2015; 2015:150325.

16.Hudson BI, Dong C, Gardener H, Elkind M, Wright CB, Goldberg R, et al.  Serum levels of soluble receptor for advanced glycation end-products and metabolic syndrome: the Northern Manhattan Study. Metabolism 2014; 63: 1125-1130.

17.Miranda ER, Somal VS, Mey JT, Blackburn BK, Wang E, Farabi S, et al. Circulating soluble RAGE isoforms are attenuated in obese, impaired-glucose-tolerant individuals and are associated with the development of type 2 diabetes. Am J Physiol Endocrinol Metab 2017; 313:E631-E640.

18.Momma H, Niu K, Kobayashi Y, Huang C, Chujo M, Otomo A, et al. Higher serum soluble receptor for advanced glycation end product levels and lower prevalence of metabolic syndrome among Japanese adult men: a cross-sectional study. Diabetol Metab Syndr 2014; 6: 33.

19.Basta G, Sironi AM, Lazzerini G, Del Turco S, Buzzigoli E, Casolaro A, Natali A, Ferrannini E, Gastaldelli A. Circulating soluble receptor for advanced glycation end products is inversely associated with glycemic control and S100A12 protein. J Clin Endocrinol Metab 2006; 91:4628-4634.

20.Huang M, Que Y, Shen X. Correlation of the plasma levels of soluble RAGE and endogenous secretory RAGE with oxidative stress in pre-diabetic patients. J Diabetes Complications 2015; 29:422-426.

21.Lee AC, Lam JK, Shiu SW, Wong Y, Betteridge DJ, Tan KC. Serum level of soluble receptor for advanced glycation end products is associated with a disintegrin and metalloproteinase 10 in type 1 diabetes. PLoS One 2015; 10:e0137330.

22.Lam JK, Wang Y, Shiu SW, Wong Y, Betteridge DJ, Tan KC. Effect of insulin on the soluble receptor for advanced glycation end products (RAGE). Diabet Med 2013; 30:702-709.

23.Yilmaz Y, Ulukaya E, Gul OO, Arabul M, Gul CB, Atug O, et al. Decreased plasma levels of soluble receptor for advanced glycation endproducts (sRAGE) in patients with nonalcoholic fatty liver disease. Clin Biochem 2009; 42:802-807.

24.Sourris KC, Harcourt BE, Penfold SA, Yap FYT, Morley AL, Morgan PE, et al. Modulation of the cellular expression of circulating advanced glycation end-product receptors in type 2 diabetic nephropathy. Exp Diabetes Res 2010; 2010:974681.

25.Uribarri J, Cai W, Woodward M, Tripp E, Goldberg L, Pyzik R, et al. Elevated serum advanced glycation endproducts in obese indicate risk for the metabolic syndrome: a link between healthy and unhealthy obesity?. J Clin Endocrinol Metab 2015; 100:1957-1966.

26.Gano LB, Donato AJ, Pierce GL, Pasha HM, Magerko KA, Roeca C, et al. Increased proinflammatory and oxidant gene expression in circulating mononuclear cells in older adults: amelioration by habitual exercise. Physiol Genomics 2011; 43:895-902.

27.Mozzini C, Garbin U, Stranieri C, Pasini A, Solani E, Tinelli IA, et al. Endoplasmic reticulum stress and Nrf2 repression in circulating cells of type 2 diabetic patients without the recommended glycemic goals. Free Radic Res 2015; 49:244-252.

28.Jiménez-Osorio AS, Picazo A, González-Reyes S, Barrera-Oviedo D, Rodríguez-Arellano ME, Pedraza-Chaverri J. Nrf2 and redox status in prediabetic and diabetic patients. Int J Mol Sci 2014; 15:20290-20305.

29.Sampath C, Rashid MR, Sang S, Ahmedna M. Specific bioactive compounds in ginger and apple alleviate hyperglycemia in mice with high fat diet-induced obesity via Nrf2 mediated pathway. Food Chem 2017; 226:79-88.

30.Hudson BI, Carter AM, Harja E, Kalea AZ, Arriero M, Yang H, et al. Identification, classification, and expression of RAGE gene splice variants. FASEB J 2008; 22:1572-1580.

31.Ryder E, Diez-Ewald M, Mosquera J, Fernández E, Pedreañez A, Vargas R, et al. Association of obesity with leukocyte count in obese individuals without metabolic syndrome. Diabetes Metab Syndr 2014; 8:197-204.

32.Durning SP, Preston-Hurlburt P, Clark PR, Xu D, Herold KC, Type 1 Diabetes Trial Net Study Group. The receptor for aAdvanced glycation endproducts (RAGE) drives T cell survival and inflammation in type 1 diabetes mellitus. J Immunol 2016; 197:3076–3085.

33.Subramanian S, Pallati PK, Sharma P, Agrawal DK, Nandipati KC. Significant association of TREM-1 with HMGB1, TLRs and RAGE in the pathogenesis of insulin resistance in obese diabetic populations. Am J Transl Res 2017; 9: 3224-3244.

34.Popa I, Ganea E, Petrescu SM. Expression and subcellular localization of RAGE in melanoma cells. Biochem Cell Biol 2014; 92:127-136.

35.Hadding A, Kaltschmidt B, Kaltschmidt C. Overexpression of receptor of advanced glycation end products hypersensitizes cells for amyloid beta peptide-induced cell death. Biochim Biophys Acta 2004; 1691:67-72.

36.Akirav EM, Preston-Hurlburt P, Garyu J, Henegariu O, Clynes R, Schmidt AM, et al. RAGE expression in human T cells: A link between environmental factors and adaptive immune responses. PLoS One 2012; 7:e34698.

37.Zhang J, Zhang L, Zhang S, Yu Q, Xiong F, Huang K, et al. HMGB1, an innate alarmin, plays a critical role in chronic inflammation of adipose tissue in obesity. Mol Cell Endocrinol 2017; 454:103-111.

38.Ridley AJ. Rho GTPases and actin dynamics in membrane protrusions and vesicle trafficking. Trends Cell Biol 2006; 16:522-529.

39.Puls A, Eliopoulos AG., Nobes CD, Bridges T, Young LS, HaIl A. Activation of the small GTPase Cdc42 by the inflammatory cytokines TNF(alpha) and IL-1, and by the Epstein-Barr virus transforming protein LMP1. J Cell Sci 1999; 112:2983-2992.