Evaluation of Tnf-α and Il-6 mRNAs expressions in visceral and subcutaneous adipose tissues of polycystic ovarian rats and effects of resveratrol

Document Type : Original Article

Authors

1 Department of Physiology, Faculty of Biological Sciences and Technology, Shahid Beheshti University, Tehran, Iran

2 Department of Biology, Faculty of Sciences, Razi University, Kermanshah, Iran

Abstract

Objective(s): Some studies suggest that chronic low-grade inflammation is involved in insulin resistance in polycystic ovary syndrome (PCOS). This study assessed possible involvement of alteration in expression of two pro-inflammatory factors, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in adipose tissues of PCOS rats in the impairment of insulin actions. Also, effects of resveratrol as an anti-inflammatory agent were investigated.
Materials and Methods: Fifteen female Wistar rats (21 days old) were divided into three groups (n=5): Ι) Control, П) PCO-model-saline: served as PCOS rats and to induce PCOS, received subcutaneously testosterone enanthate 1 mg/100 g body weight subcutaneously for 35 days, Ш) PCO-model-resveratrol, after receiving testosterone, received resveratrol 10 mg/kg intraperitoneally for 28 days. The expression of Tnf-α and Il-6 mRNAs in adipose tissues was determined by the qRT-PCR method.
Results: The Il-6 mRNA expression in the visceral adipose tissue of PCOS rats was increased in comparison to controls (P<0.05). Tnf-α and Il-6 mRNA expression in visceral and subcutaneous adipose tissues of polycystic ovarian rats was similar to controls. The expression of Tnf-α mRNA in subcutaneous adipose tissue and Tnf-α and Il-6 mRNAs in the visceral adipose tissue of the PCO-model-resveratrol group were lower than PCOS rats (P<0.05).
Conclusion: Increased expression of Il-6 mRNA in the visceral adipose tissue of polycystic ovarian rats may be one cause of insulin resistance observed in them and resveratrol as an anti-inflammatory and anti-hyperglycemic agent may decrease the risk of diabetes by reduction of expression of pro-inflammatory cytokines TNF-α and IL-6 in PCOS patients.

Keywords

Main Subjects

References

1.Johansson J, Stener-Victorin E. Polycystic ovary syndrome: effect and mechanisms of acupuncture for ovulation induction. Evid Based Complement Alternat Med 2013; 2013:762615.
2. Sathyapalan T, Atkin SL. Mediators of inflammation in polycystic ovary syndrome in relation to adiposity. Mediat Inflamm 2010; 2010:758654.
3. Dunaif A. Insulin resistance and the polycystic ovary syndrome: mechanism and implications for pathogenesis. Endocr Rev 1997;18:774-800.
4.Desai V, Prasad NR, Manohar SM, Sachan A, Narasimha SRPVL, Bitla ARR. Oxidative stress in non-obese women with polycystic ovarian syndrome. J Clin Diagn Res 2014;8:CC01-CC03.
5.Escobar-Morreale HF, Luque-Ramírez M, González F. Circulating inflammatory markers in polycystic ovary syndrome: a systematic review and metaanalysis. Fertil Steril 2011;95:1048-1058. e1-2.
6.Rojas J, Chávez M, Olivar L, Rojas M, Morillo J, Mejías J, et al. Polycystic ovary syndrome, insulin resistance, and obesity: navigating the pathophysiologic labyrinth. Int J Reprod Med 2014; 2014:719050.
7.Xiong Y-l, Liang X-y, Yang X, Li Y, Wei L-n. Low-grade chronic inflammation in the peripheral blood and ovaries of women with polycystic ovarian syndrome. Euro J Ob Gyn Repro Biol 2011;159:148-150.
8.González F. Inflammation in polycystic ovary syndrome: underpinning of insulin resistance and ovarian dysfunction. Steroids 2012;77:300-305.
9.Zahorska-Markiewicz B, Janowska J, Olszanecka-Glinianowicz M, Zurakowski A. Serum concentrations of TNF-[alpha] and soluble TNF-[alpha] receptors in obesity. Int J Obes Relat Metab Disord 2000;24:1392-1395.
10.Hofmann C, Lorenz K, Braithwaite S, Colca J, Palazuk B, Hotamisligil G, et al. Altered gene expression for tumor necrosis factor-alpha and its receptors during drug and dietary modulation of insulin resistance. Endocrinology 1994;134:264-270.
11.Kern PA, Ranganathan S, Li C, Wood L, Ranganathan G. Adipose tissue tumor necrosis factor and interleukin-6 expression in human obesity and insulin resistance. Am J Physiol Endocrinol Metab 2001;280:E745-E751.
12.Mohlig M, Spranger J, Osterhoff M, Ristow M, Pfeiffer A, Schill T, et al. The polycystic ovary syndrome per se is not associated with increased chronic inflammation. Eur J Endocrinol 2004;150:525-532.
13.Lin YS, Tsai SJ, Lin MW, Yang CT, Huang MF, Wu MH. Interleukin‐6 as an early chronic inflammatory marker in polycystic ovary Syndrome with insulin receptor substrate‐2 polymorphism. Am J Reprod Immunol 2011;66:527-533.
14.Escobar‐Morreale HF, Calvo RM, Villuendas G, Sancho J, Millán JL. Association of polymorphisms in the interleukin 6 receptor complex with obesity and hyperandrogenism. Obes Res 2003;11:987-996.
15.González F, Nair KS, Daniels JK, Basal E, Schimke JM, Blair HE. Hyperandrogenism sensitizes leukocytes to hyperglycemia to promote oxidative stress in lean reproductive-age women. J Clin Endocrinol Metab 2012;97:2836-2843.
16.Franckhauser S, Elias I, Sopasakis VR, Ferre T, Nagaev I, Andersson CX, et al. Overexpression of Il6 leads to hyperinsulinaemia, liver inflammation and reduced body weight in mice. Diabetologia 2008;51:1306-1316.
17.Rui L, Aguirre V, Kim JK, Shulman GI, Lee A, Corbould A, et al. Insulin/IGF-1 and TNF-α stimulate phosphorylation of IRS-1 at inhibitory Ser 307 via distinct pathways. J Clin Invest 2001;107:181-189.
18.Holmes AG, Mesa JL, Neill BA, Chung J, Carey AL, Steinberg GR, et al. Prolonged interleukin-6 administration enhances glucose tolerance and increases skeletal muscle PPARα and UCP2 expression in rats. J Endocrinol 2008;198:367-374.
19.Ciaraldi TP, el-Roeiy A, Madar Z, Reichart D, Olefsky JM, Yen S. Cellular mechanisms of insulin resistance in polycystic ovarian syndrome. J Clin Endocrinol Metab 1992; 75: 577-583.
20.Dunaif A, Wu X, Lee A, Diamanti-Kandarakis E. Defects in insulin receptor signaling in vivo in the polycystic ovary syndrome (PCOS). Am J Physiol Endocrinol Metab 2001;281:E392-E399.
21.Griffin ME, Marcucci MJ, Cline GW, Bell K, Barucci N, Lee D, et al. Free fatty acid-induced insulin resistance is associated with activation of protein kinase C theta and alterations in the insulin signaling cascade. Diabetes 1999;48:1270-1274.
22.Yu C, Chen Y, Cline GW, Zhang D, Zong H, Wang Y, et al. Mechanism by which fatty acids inhibit insulin activation of insulin receptor substrate-1 (IRS-1)-associated phosphatidylinositol 3-kinase activity in muscle. J Biol Chem 2002;277:50230-50236.
23.Tanti J-F, Gremeaux T, Van Obberghen E, Le Marchand-Brustel Y. Serine/threonine phosphorylation of insulin receptor substrate 1 modulates insulin receptor signaling. J Biol Chem 1994; 269: 6051-6057.
24.Brasnyó P, Molnár GA, Mohás M, Markó L, Laczy B, Cseh J, et al. Resveratrol improves insulin sensitivity, reduces oxidative stress and activates the Akt pathway in type 2 diabetic patients. Br J Nutr 2011;106:383-389.
25.Gonzalez F, Thusu K, Abdel-Rahman E, Prabhala A, Tomani M, Dandona P. Elevated serum levels of tumor necrosis factor alpha in normal-weight women with polycystic ovary syndrome. Metabolism 1999;48:437-441.
26.Escobar-Morreale H, Villuendas G, Botella-Carretero J, Sancho J, San Millan J. Obesity, and not insulin resistance, is the major determinant of serum inflammatory cardiovascular risk markers in pre-menopausal women. Diabetologia 2003;46:625-633.
27.Beloosesky R, Gold R, Almog B, Sasson R, Dantes A, Land-Bracha A, et al. Induction of polycystic ovary by testosterone in immature female rats: modulation of apoptosis and attenuation of glucose/insulin ratio. Int J Mol Med 2004;14:207-216.
28.Ergenoglu M, Yildirim N, Yildirim AGS, Yeniel O, Erbas O, Yavasoglu A, et al. Effects of resveratrol on ovarian morphology, plasma anti-mullerian hormone, IGF-1 levels, and oxidative stress parameters in a rat model of polycystic ovary syndrome. Reprod Sci 2015;22:942-947.
29.Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative CT method. Nat Protoc 2008;3:1101-1108.
30.Aref A-BM, Ahmed OM, Ali LA, Semmler M. Maternal rat diabetes mellitus deleteriously affects insulin sensitivity and Beta-cell function in the offspring. J Diabetes Res 2013;2013.
31.González F, Rote NS, Minium J, Kirwan JP. Reactive oxygen species-induced oxidative stress in the development of insulin resistance and hyperandrogenism in polycystic ovary syndrome. Clin Endocrinol Metab 2006;91:336-340.
32.O'Driscoll J, Mamtora H, Higginson J, Pollock A, Kane J, Anderson D. A prospective study of the prevalence of clear‐cut endocrine disorders and polycystic ovaries in 350 patients presenting with hirsutism or androgenic alopecia.. Clin Endocrinol (Oxf) 1994;41:231-236.
33.Horejsi R, Möller R, Rackl S, Giuliani A, Freytag U, Crailsheim K, et al. Android subcutaneous adipose tissue topography in lean and obese women suffering from PCOS: comparison with type 2 diabetic women. Am J Phys Anthropol 2004;124:275-281.
34.Goodarzi MO, Korenman SG. The importance of insulin resistance in polycystic ovary syndrome. Fertil Steril 2003;80:255-258.
35.ESHRE TR, Group A-SPCW. Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome. . Fertil Steril 2004;81:19-25.
36.Hotamisligil GS, Shargill NS, Spiegelman BM. Adipose Expression of Tumor Necrosis Factor- : Direct role in obesity-linked insulin resistance. Science 1993;259:87-91.
37.Gwechenberger M, Mendoza LH, Youker KA, Frangogiannis NG, Smith CW, Michael LH, et al. Cardiac myocytes produce interleukin-6 in culture and in viable border zone of reperfused infarctions. Circulation 1999;99:546-551.
38.Orio Jr F, Palomba S, Cascella T, Milan G, Mioni R, Pagano C, et al. Adiponectin levels in women with polycystic ovary syndrome. J Clin Endocrinol Metab 2003;88:2619-2623.
39.Fulghesu AM, Sanna F, Uda S, Magnini R, Portoghese E, Batetta B. IL-6 serum levels and production is related to an altered immune response in polycystic ovary syndrome girls with insulin resistance. Mediators Inflamm 2011;2011:389317.
40.Heutling D, Schulz H, Nickel I, Kleinstein J, Kaltwasser P, Krzyzanowska K, et al. Endothelial, inflammatory and endocrine markers in women with PCOS before and after metformin treatment. Exp Clin Endocrinol Diabetes 2006;114: 15_195.
41.Raison CL, Capuron L, Miller AH. Cytokines sing the blues: inflammation and the pathogenesis of depression. Trends Immunol 2006;27:24-31.
42.Stephens J, Pekala P. Transcriptional repression of the C/EBP-alpha and GLUT4 genes in 3T3-L1 adipocytes by tumor necrosis factor-alpha. Regulations is coordinate and independent of protein synthesis. J Biol Chem 1992;267:13580-13584.
43.Boden G. Role of fatty acids in the pathogenesis of insulin resistance and NIDDM. Diabetes 1997;46:3-10.
44.Teruel T, Hernandez R, Lorenzo M. Ceramide mediates insulin resistance by tumor necrosis factor-α in brown adipocytes by maintaining Akt in an inactive dephosphorylated state. Diabetes 2001;50:2563-2571.
45.Shimabukuro M, Zhou Y-T, Levi M, Unger RH. Fatty acid-induced β cell apoptosis: a link between obesity and diabetes. Proc Natl Acad Sci U S A 1998;95:2498-2502.
46.Barnes PJ, Karin M. Nuclear factor-κB—a pivotal transcription factor in chronic inflammatory diseases. N Engl J Med 1997;336:1066-1071.
47.Côté CD, Rasmussen BA, Duca FA, Zadeh-Tahmasebi M, Baur JA, Daljeet M, et al. Resveratrol activates duodenal Sirt1 to reverse insulin resistance in rats through a neuronal network. Nat Med 2015;21:498-505.
48.Baur JA, Sinclair DA. Therapeutic potential of resveratrol: the in vivo evidence. Nat Rev Drug Discov 2006;5:493-506.
49.Zheng X, Zhu S, Chang S, Cao Y, Dong J, Li J, et al. Protective effects of chronic resveratrol treatment on vascular inflammatory injury in streptozotocin-induced type 2 diabetic rats: Role of NF-kappa B signaling. Eur J Pharmacol 2013;720:147-157.
50.Su H-C, Hung L-M, Chen J-K. Resveratrol, a red wine antioxidant, possesses an insulin-like effect in streptozotocin-induced diabetic rats. Am J Physiol Endocrinol Metab 2006;290:E1339-E1346.
51.Soufi FG, Mohammad-Nejad D, Ahmadieh H. Resveratrol improves diabetic retinopathy possibly through oxidative stress–nuclear factor κB–apoptosis pathway. Pharmacol Rep 2012;64:1505-1514.
52.Chanock SJ, El Benna J, Smith RM, Babior BM. The respiratory burst oxidase. J Biol Chem 1994;270:24519-2422.
53.Sharma S, Anjaneyulu M, Kulkarni S, Chopra K. Resveratrol, a polyphenolic phytoalexin, attenuates diabetic nephropathy in rats. Pharmacology 2006;76:69-75.
54. Kumar A, Kaundal RK, Iyer S, Sharma SS. Effects of resveratrol on nerve functions, oxidative stress and DNA fragmentation in experimental diabetic neuropathy. Life Sci 2007;8013:1236-1244.
Iranian Journal of Basic Medical Sciences
Volume 21, Issue 2
February 2018
Pages 165-174

Files

Share

How to cite

Statistics