Enhanced Inflammation without Impairment of Insulin Signaling in the Visceral Adipose Tissue of 5α-Dihydrotestosterone-Induced Animal Model of Polycystic Ovary Syndrome

Danijela Vojnović Milutinović; Marina Nikolić; Nataša Veličković; Ana Djordjevic; Biljana Bursać; Jelena Nestorov; Ana Teofilović; Ivana Božić Antić; Jelica Bjekić Macut; Abdulbaset Shirif Zidane; Gordana Matić; Djuro Macut

doi:10.1055/s-0043-104531

Experimental and Clinical Endocrinology & Diabetes, Table of Contents

Exp Clin Endocrinol Diabetes 2017; 125(08): 522-529
DOI: 10.1055/s-0043-104531

Article

Enhanced Inflammation without Impairment of Insulin Signaling in the Visceral Adipose Tissue of 5α-Dihydrotestosterone-Induced Animal Model of Polycystic Ovary Syndrome

Danijela Vojnović Milutinović

¹Department of Biochemistry, Institute for Biological Research “Siniša Stanković”, University of Belgrade, Belgrade, Serbia

,

Marina Nikolić

¹Department of Biochemistry, Institute for Biological Research “Siniša Stanković”, University of Belgrade, Belgrade, Serbia

,

Nataša Veličković

¹Department of Biochemistry, Institute for Biological Research “Siniša Stanković”, University of Belgrade, Belgrade, Serbia

,

Ana Djordjevic

¹Department of Biochemistry, Institute for Biological Research “Siniša Stanković”, University of Belgrade, Belgrade, Serbia

,

Biljana Bursać

¹Department of Biochemistry, Institute for Biological Research “Siniša Stanković”, University of Belgrade, Belgrade, Serbia

,

Jelena Nestorov

¹Department of Biochemistry, Institute for Biological Research “Siniša Stanković”, University of Belgrade, Belgrade, Serbia

,

Ana Teofilović

¹Department of Biochemistry, Institute for Biological Research “Siniša Stanković”, University of Belgrade, Belgrade, Serbia

,

Ivana Božić Antić

²Clinic of Endocrinology, Diabetes and Metabolic Diseases, Clinical Center of Serbia and Faculty of Medicine, University of Belgrade, Belgrade, Serbia

,

Jelica Bjekić Macut

³UMC Bežanijska kosa, Bežanijska kosa bb, Faculty of Medicine, University of Belgrade, Belgrade, Serbia

,

Abdulbaset Shirif Zidane

¹Department of Biochemistry, Institute for Biological Research “Siniša Stanković”, University of Belgrade, Belgrade, Serbia

,

Gordana Matić

¹Department of Biochemistry, Institute for Biological Research “Siniša Stanković”, University of Belgrade, Belgrade, Serbia

,

Djuro Macut

²Clinic of Endocrinology, Diabetes and Metabolic Diseases, Clinical Center of Serbia and Faculty of Medicine, University of Belgrade, Belgrade, Serbia

› Author Affiliations

Abstract

Polycystic ovary syndrome is a heterogeneous endocrine and metabolic disorder associated with abdominal obesity, dyslipidemia and insulin resistance. Since abdominal obesity is characterized by low-grade inflammation, the aim of the study was to investigate whether visceral adipose tissue inflammation linked to abdominal obesity and dyslipidemia could lead to impaired insulin sensitivity in the animal model of polycystic ovary syndrome.

Female Wistar rats were treated with nonaromatizable 5α-dihydrotestosterone pellets in order to induce reproductive and metabolic characteristics of polycystic ovary syndrome. Glucose, triglycerides, non-esterified fatty acids and insulin were determined in blood plasma. Visceral adipose tissue inflammation was evaluated by the nuclear factor kappa B intracellular distribution, macrophage migration inhibitory factor protein level, as well as TNFα, IL6 and IL1β mRNA levels. Insulin sensitivity was assessed by intraperitoneal glucose tolerance test and homeostasis model assessment index, and through analysis of insulin signaling pathway in the visceral adipose tissue.

Dihydrotestosterone treatment led to increased body weight, abdominal obesity and elevated triglycerides and non-esterified fatty acids, which were accompanied by the activation of nuclear factor kappa B and increase in macrophage migration inhibitory factor, IL6 and IL1β levels in the visceral adipose tissue. In parallel, insulin sensitivity was affected in 5α-dihydrotestosterone-treated animals only at the systemic and not at the level of visceral adipose tissue.

The results showed that abdominal obesity and dyslipidemia in the animal model of polycystic ovary syndrome were accompanied with low-grade inflammation in the visceral adipose tissue. However, these metabolic disturbances did not result in decreased tissue insulin sensitivity.

Key words

PCOS - 5α-dihydrotestosterone - visceral adipose tissue - inflammation - insulin resistance

Full Text

References

References
1 Legro RS, Arslanian SA, Ehrmann DA. et al. Diagnosis and treatment of polycystic ovary syndrome: An Endocrine Society clinical practice guideline. J Clin Endocrinol Metab 2013; 98: 4565-4592
2 Conway G, Dewailly D, Diamanti-Kandarakis E. et al. European survey of diagnosis and management of the polycystic ovary syndrome: Results of the ESE PCOS special interest group’s questionnaire. Eur J Endocrinol 2014; 171: 489-498
3 Diamanti-Kandarakis E, Dunaif A. Insulin resistance and the polycystic ovary syndrome revisited: An update on mechanisms and implications. Endocr Rev 2012; 33: 981-1030
4 Svendsen PF, Nilas L, Norgaard K. et al. Obesity, body composition and metabolic disturbances in polycystic ovary syndrome. Hum Reprod 2008; 23: 2113-2121
5 Jung UJ, Choi MS. Obesity and its metabolic complications: The role of adipokines and the relationship between obesity, inflammation, insulin resistance, dyslipidemia and nonalcoholic fatty liver disease. Int J Mol Sci 2014; 15: 6184-6223
6 Weisberg SP, McCann D, Desai M. et al. Obesity is associated with macrophage accumulation in adipose tissue. J Clin Invest 2003; 112: 1796-1808
7 Piya MK, McTernan PG, Kumar S. Adipokine inflammation and insulin resistance: the role of glucose, lipids and endotoxin. J Endocrinol 2013; 216: T1-T15
8 Xu A, Chan KW, Hoo RL. et al. Testosterone selectively reduces the high molecular weight form of adiponectin by inhibiting its secretion from adipocytes. J Biol Chem 2005; 280: 18073-18080
9 Gonzalez F. Inflammation in Polycystic Ovary Syndrome: Underpinning of insulin resistance and ovarian dysfunction. Steroids 2012; 77: 300-305
10 Baker RG, Hayden MS, Ghosh S. NF-kappaB, inflammation, and metabolic disease. Cell Metab 2011; 13: 11-22
11 Sanchez-Zamora YI, Rodriguez-Sosa M. The role of MIF in type 1 and type 2 diabetes mellitus. J Diabetes Res 2014; 2014: 804519
12 Lue H, Kleemann R, Calandra T. et al. Macrophage migration inhibitory factor (MIF): mechanisms of action and role in disease. Microbes Infect 2002; 4: 449-460
13 Azziz R, Carmina E, Dewailly D. et al. The Androgen Excess and PCOS Society criteria for the polycystic ovary syndrome: The complete task force report. Fertil Steril 2009; 91: 456-488
14 Ovalle F, Azziz R. Insulin resistance, polycystic ovary syndrome, and type 2 diabetes mellitus. Fertil Steril 2002; 77: 1095-1105
15 Macut D, Simic T, Lissounov A. et al. Insulin resistance in non-obese women with polycystic ovary syndrome: Relation to byproducts of oxidative stress. Exp Clin Endocrinol Diabetes 2011; 119: 451-455
16 Panidis D, Tziomalos K, Misichronis G. et al. Insulin resistance and endocrine characteristics of the different phenotypes of polycystic ovary syndrome: a prospective study. Hum Reprod 2012; 27: 541-549
17 Pirola L, Johnston AM, Van Obberghen E. Modulation of insulin action. Diabetologia 2004; 47: 170-184
18 Fassnacht M, Schlenz N, Schneider SB. et al. Beyond adrenal and ovarian androgen generation: Increased peripheral 5 alpha-reductase activity in women with polycystic ovary syndrome. J Clin Endocrinol Metab 2003; 88: 2760-2766
19 Silfen ME, Denburg MR, Manibo AM. et al. Early endocrine, metabolic, and sonographic characteristics of polycystic ovary syndrome (PCOS): Comparison between nonobese and obese adolescents. J Clin Endocrinol Metab 2003; 88: 4682-4688
20 Manneras L, Cajander S, Holmang A. et al. A new rat model exhibiting both ovarian and metabolic characteristics of polycystic ovary syndrome. Endocrinology 2007; 148: 3781-3791
21 Yanes LL, Romero DG, Moulana M. et al. Cardiovascular-renal and metabolic characterization of a rat model of polycystic ovary syndrome. Gend Med 2011; 8: 103-115
22 Sara L, Antal P, Masszi G. et al. Arteriolar insulin resistance in a rat model of polycystic ovary syndrome. Fertil Steril 2012; 97: 462-468
23 Nikolić M, Macut D, Djordjevic A. et al. Possible involvement of glucocorticoids in 5alpha-dihydrotestosterone-induced PCOS-like metabolic disturbances in the rat visceral adipose tissue. Mol Cell Endocrinol 2015; 399: 22-31
24 Duncombe WG. The Colorimetric Micro-Determination of Non-Esterified Fatty Acids in Plasma. Clin Chim Acta 1964; 9: 122-125
25 Spector T. Refinement of the coomassie blue method of protein quantitation. A simple and linear spectrophotometric assay for less than or equal to 0.5 to 50 microgram of protein. Anal Biochem 1978; 86: 142-146
26 Veličković N, Djordjevic A, Vasiljević A. et al. Tissue-specific regulation of inflammation by macrophage migration inhibitory factor and glucocorticoids in fructose-fed Wistar rats. Br J Nutr 2013; 110: 456-465
27 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25: 402-408
28 Macut D, Panidis D, Glisic B. et al. Lipid and lipoprotein profile in women with polycystic ovary syndrome. Can J Physiol Pharmacol 2008; 86: 199-204
29 Kelly CC, Lyall H, Petrie JR. et al. Low grade chronic inflammation in women with polycystic ovarian syndrome. J Clin Endocrinol Metab 2001; 86: 2453-2455
30 Escobar-Morreale HF, Villuendas G, Botella-Carretero JI. et al. Obesity, and not insulin resistance, is the major determinant of serum inflammatory cardiovascular risk markers in pre-menopausal women. Diabetologia 2003; 46: 625-633
31 Knebel B, Janssen OE, Hahn S. et al. Increased low grade inflammatory serum markers in patients with Polycystic ovary syndrome (PCOS) and their relationship to PPARgamma gene variants. Exp Clin Endocrinol Diabetes 2008; 116: 481-486
32 Mohlig M, Spranger J, Osterhoff M. et al. The polycystic ovary syndrome per se is not associated with increased chronic inflammation. Eur J Endocrinol 2004; 150: 525-532
33 Auron PE. The interleukin 1 receptor: Ligand interactions and signal transduction. Cytokine Growth Factor Rev 1998; 9: 221-237
34 O’Neill LA, Greene C. Signal transduction pathways activated by the IL-1 receptor family: Ancient signaling machinery in mammals, insects, and plants. J Leukoc Biol 1998; 63: 650-657
35 Yan S, Xu Z, Lou F. et al. NF-kappaB-induced microRNA-31 promotes epidermal hyperplasia by repressing protein phosphatase 6 in psoriasis. Nat Commun 2015; 6: 7652
36 Dali-Youcef N, Mecili M, Ricci R. et al. Metabolic inflammation: Connecting obesity and insulin resistance. Ann Med 2013; 45: 242-253
37 Suganami T, Tanimoto-Koyama K, Nishida J. et al. Role of the Toll-like receptor 4/NF-kappaB pathway in saturated fatty acid-induced inflammatory changes in the interaction between adipocytes and macrophages. Arterioscler Thromb Vasc Biol 2007; 27: 84-91
38 Finucane OM, Reynolds CM, McGillicuddy FC. et al. Insights into the role of macrophage migration inhibitory factor in obesity and insulin resistance. Proc Nutr Soc 2012; 71: 622-633
39 Verschuren L, Kooistra T, Bernhagen J. et al. MIF deficiency reduces chronic inflammation in white adipose tissue and impairs the development of insulin resistance, glucose intolerance, and associated atherosclerotic disease. Circ Res 2009; 105: 99-107
40 Luque-Ramirez M, Escobar-Morreale HF. Polycystic ovary syndrome as a paradigm for prehypertension, prediabetes, and preobesity. Curr Hypertens Rep 2014; 16: 500
41 Randeva HS, Tan BK, Weickert MO. et al. Cardiometabolic aspects of the polycystic ovary syndrome. Endocr Rev 2012; 33: 812-841
42 Shoelson SE, Lee J, Goldfine AB. Inflammation and insulin resistance. J Clin Invest 2006; 116: 1793-1801
43 Jager J, Gremeaux T, Cormont M. et al. Interleukin-1beta-induced insulin resistance in adipocytes through down-regulation of insulin receptor substrate-1 expression. Endocrinology 2007; 148: 241-251
44 Vozarova B, Weyer C, Hanson K. et al. Circulating interleukin-6 in relation to adiposity, insulin action, and insulin secretion. Obes Res 2001; 9: 414-417
45 Tepavčević S, Vojnović Milutinović D, Macut D. et al. Dihydrotestosterone deteriorates cardiac insulin signaling and glucose transport in the rat model of polycystic ovary syndrome. J Steroid Biochem Mol Biol 2014; 141: 71-76
46 Khodabandehloo H, Gorgani-Firuzjaee S, Panahi G. et al. Molecular and cellular mechanisms linking inflammation to insulin resistance and beta-cell dysfunction. Transl Res 2016; 167: 228-256