Testosterone enhances lipopolysaccharide-induced interleukin-6 and macrophage chemotactic protein-1 expression by activating the extracellular signal-regulated kinase 1/2/nuclear factor-κB signalling pathways in 3T3-L1 adipocytes

Su,Chunlin; Chen,Min; Huang,Haiyan; Lin,Jinfang

doi:10.3892/mmr.2015.3401

Testosterone enhances lipopolysaccharide-induced interleukin-6 and macrophage chemotactic protein-1 expression by activating the extracellular signal-regulated kinase 1/2/nuclear factor-κB signalling pathways in 3T3-L1 adipocytes

Authors:
- Chunlin Su
- Min Chen
- Haiyan Huang
- Jinfang Lin
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Affiliations: Department of Gynecology, Shanghai Obstetrics and Gynecology Hospital Affiliated to Fudan University, Shanghai 200011, P.R. China, Department of Reproductive Medicine Center, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, P.R. China, Key Laboratory of Molecular Medicine Ministry of Education, Shanghai Medical University Affliated to Fudan University, Shanghai 200011, P.R. China
Published online on: March 3, 2015 https://doi.org/10.3892/mmr.2015.3401
Pages: 696-704

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Abstract

Low‑grade chronic inflammation is commonly found in patients with polycystic ovary syndrome (PCOS) who exhibit hyperandrogenism or hyperandrogenemia. Clinical studies have shown that hyperandrogenemia is closely correlated with low‑grade chronic inflammation. However, the mechanism underlying this correlation remains unclear. Recent studies have suggested that adipocytes increase the production of proinflammatory mediators such as interleukin‑6 (IL‑6) and macrophage chemotactic protein‑1 (MCP‑1) when the inflammatory signal transduction cascade system is activated by external stimuli. The present study aimed to evaluate the effects of testosterone on the innate signalling and expression of proinflammatory mediators in 3T3‑L1 adipocytes, which were or were not induced by lipopolysaccharide (LPS). The effects of testosterone on the expression of proinflammatory mediators, nuclear factor‑κB (NF‑κB), and extracellular signal‑regulated kinase 1/2 (ERK1/2) signalling pathways were investigated using an enzyme‑linked immunosorbent assay, reverse transcriptase‑polymerase chain reaction, western blot analysis and an electrophoresis mobility shift assay. Testosterone induces IL‑6 and MCP‑1, and enhances LPS‑induction of IL‑6 and MCP‑1. However, the effects are not simply additive, testosterone significantly enhanced the effects of LPS‑induced inflammation factors. Testosterone induces the phosphorylation of ERK1/2 and NF‑κB. The effect of testosterone on the expression of IL‑6 and MCP‑1 is inhibited by PD98059 , an ERK1/2 inhibitor, and PDTC, an NF‑κB inhibitor. The results indicate that testosterone enhances LPS‑induced IL‑6 and MCP‑1 expression by activating the ERK1/2/NF‑κB signalling pathways in 3T3‑L1 adipocytes.

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1	Legro RS, Kunselman AR, Dodson WC and Dunaif A: Prevalence and predictors of risk for type 2 diabetes mellitus and impaired glucose tolerance in polycystic ovary syndrome: a prospective, controlled study in 254 affected women. J Clin Endocrinol Metab. 84:165–169. 1999.PubMed/NCBI
2	Hu G, Qiao Q, Tuomilehto J, Balkau B, Borch-Johnsen K and Pyorala K; DECODE Study Group: Prevelence of the metabolic syndrome and its relation to all-cause and cardiovascular mortality in nondiabetic European men and women. Arch Intern Med. 164:1066–1076. 2004. View Article : Google Scholar : PubMed/NCBI
3	Boura-Halfon S and Zick Y: Phosphorylation of IRS proteins, insulin action, and insulin resistance. Am J Physiol Endocrinol Metab. 296:E581–E591. 2009. View Article : Google Scholar
4	Coviello AD, Legro RS and Dunaif A: Adolescent girls with polycystic ovary syndrome have an increased risk of the metabolic syndrome associated with increasing androgen levels independent of obesity and insulin resistance. J Clin Endocrinol Metab. 91:492–497. 2006. View Article : Google Scholar
5	Banaszewska B, Pawelczyk L, Spaczynski RZ, Dziura J and Duleba AJ: Effects of simvastatin and oral contraceptive agent on polycystic ovary syndrome: prospective, randomized, crossover trial. J Clin Endocrinol Metab. 92:456–461. 2007. View Article : Google Scholar
6	Ibáñez L, Jaramillo AM, Ferrer A and de Zegher F: High neutrophil count in girls and women with hyperinsulinaemic hyperandrogenism: normalization with metformin and flutamide overcomes the aggravation by oral contraception. Hum Reprod. 20:2457–2462. 2005. View Article : Google Scholar : PubMed/NCBI
7	González F, Rote NS, Minium J and Kirwan JP: In vitro evidence that hyperglycemia stimulates tumor necrosis factor-alpha release in obese women with polycystic ovary syndrome. J Endocrinol. 188:521–529. 2006. View Article : Google Scholar : PubMed/NCBI
8	Tataranni PA and Ortega E: A burning question: does an adipokine-induced activation of the immune system mediate the effect of overnutrition on type 2 diabetes? Diabetes. 54:917–927. 2005. View Article : Google Scholar : PubMed/NCBI
9	Lumeng CN and Saltiel AR: Inflammatory links between obesity and metabolic disease. J Clin Invest. 121:2111–2117. 2011. View Article : Google Scholar : PubMed/NCBI
10	Nicklas BJ, Ambrosius W, Messier SP, et al: Diet-induced weight loss, exercise, and chronic inflammation in older, obese adults: a randomized controlled clinical trial. Am J Clin Nutr. 79:544–551. 2004.PubMed/NCBI
11	Mohamed-Ali V, Goodrick S, Rawesh A, et al: Subcutaneous adipose tissue releases interleukin-6, but not tumour necrosis factor-alpha, in vivo. J Clin Endocrinol Metab. 82:4196–4200. 1997.PubMed/NCBI
12	Lagathu C, Bastard JP, Auclair M, Maachi M, Capeau J and Caron M: Chronic interleukin (IL-6) treatment increased IL-6 secretion and induced insulin resistance in adipocyte: prevetion by rosigilitazone. Biochem Biophys Res Commun. 311:372–379. 2003. View Article : Google Scholar : PubMed/NCBI
13	Homaidan FR, Chakroun I and El-Sabban ME: Regulation of nuclear factor-kappaB in intestinal epithelial cells in a cell model inflammation. Mediators Inflamm. 12:277–283. 2003. View Article : Google Scholar
14	Shoelson SE, Lee J and Goldfine AB: Inflammation and insulin resistance. Clin Invest. 116:1793–1801. 2006. View Article : Google Scholar
15	Ognjanovic S, Jacobs DR, Steinberger J, Moran A and Sinako AR: Relation of chemokines to BMI and insulin resistance at ages 18–21. Int J Obes (Lond). 37:420–423. 2013. View Article : Google Scholar
16	Gerhardt CC, Romero IA, Cancello R, Camoin L and Strosberg AD: Chemokines control fat accumulation and leptin secretion by cultured human adipocytes. Mol Cell Endocrinol. 175:81–92. 2001. View Article : Google Scholar : PubMed/NCBI
17	Takahashi K, Mizuarai S, Araki H, et al: Adipocity elevates plasma MCP-1 levels leading to the increased CD11b-positive monocytes in mice. J Biol Chem. 278:46654–46660. 2003. View Article : Google Scholar : PubMed/NCBI
18	Kershaw EE and Flier JS: Adipose tissue as an endocrine organ. J Clin Endocrinol Metab. 89:2548–2556. 2004. View Article : Google Scholar : PubMed/NCBI
19	Hsiung SC, Tamir H, Franke TF and Liu KP: Roles of extracellular signal-regulated kinase and Akt signaling in coordinating nuclear transcription factor-kappaB-dependent cell survival after serotonin 1A receptor activation. J Neurochem. 95:1653–1666. 2005. View Article : Google Scholar : PubMed/NCBI
20	Kaidashev IP: Activation of NF-κB under metabolic syndrome. Internationl J Physiol Pathophysiol. 3:287–297. 2012. View Article : Google Scholar
21	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. 27:84–91. 2007. View Article : Google Scholar
22	Kovacheva EL, Hikim AP, Shen R, Sinha I and Sinha-Hikim I: Testosterone supplementation reverses sarcopenia in aging through regulation of myostatin, c-Jun NH2-terminal kinase, notch, and Akt signaling pathways. Endocrinology. 151:628–638. 2010. View Article : Google Scholar :
23	Sun HZ, Yang TW, Zang WJ and Wu SF: Dehydroepiandrosterone-induced proliferation of prostatic epithelial cell is mediated by NFKB via PI3K/AKT signaling pathway. J Endocrinol. 204:311–318. 2010. View Article : Google Scholar
24	Wu Y, Bauman WA, Blitzer RD and Cardozo C: Testosterone- induced hypertrophy of L6 myoblasts is dependent upon Erk and mTOR. Biochem Biophys Res Commun. 400:679–683. 2010. View Article : Google Scholar : PubMed/NCBI
25	Yamamoto Y, Yoshimasa Y, Koh M, et al: Constitutively active mitogen-activated protein kinase kinase increases GLUT1 expression and recruits both GLUT1 and GLUT4 at the cell surface in 3T3-L1 adipocytes. Diabetes. 49:332–339. 2000. View Article : Google Scholar : PubMed/NCBI
26	Yin J, Zuberi A, Gao Z, Liu D, Liu Z and Ye J: Shilianhua extract inhibits GSK-3beta and promotes glucose metabolism. Am J Physiol Endocrinol Metab. 296:E1275–E1280. 2009. View Article : Google Scholar : PubMed/NCBI
27	Benson S, Janssen OE, Hahn S, et al: Obesity, depression, and chronic low-grade inflammation in women with polycystic ovary syndrome. Brain Behav Immun. 22:177–184. 2008. View Article : Google Scholar
28	Marino JS, Iler J, Dowling AR, et al: Adipocyte dysfunction in a mouse model of polycystic ovary syndrome (PCOS): evidence of adipocyte hypertrophy and tissue-specific inflammation. PLoS One. 7:e486432012. View Article : Google Scholar : PubMed/NCBI
29	Greenberg AS and Obin MS: Obesity and the role of adipose tissue in inflammation and metabolism. Am J Clin Nutr. 83:461S–465S. 2006.PubMed/NCBI
30	Lei Y, Zhang Y, Cao Y, et al: Up-regulation of bradykinin receptors in rat bronchi via I kappa B kinase-mediated inflammatory signaling pathway. Eur J Pharmacol. 634:149–161. 2010. View Article : Google Scholar : PubMed/NCBI
31	Corbould A, Zhao H, Mirzoeva S, Aird F and Dunaif A: Enhanced mitogenic signaling in skeletal muscle of women with polycystic ovary syndrome. Diabetes. 55:751–759. 2006. View Article : Google Scholar : PubMed/NCBI
32	González F, Rote NS, Minium J and Kirwan JP: Increased activation of nuclear factor kappaB triggers inflammation and insulin resistance in polycystic ovary syndrome. J Clin Endocrinol Metab. 91:1508–1512. 2006. View Article : Google Scholar : PubMed/NCBI
33	Yeung YT, Bryce NS, Adams S, et al: p38 MAPK inhibitors attenuate pro-inflammatory cytokine production and the inva-siveness of human U251 glioblastoma cells. J Neurooncol. 109:35–44. 2012. View Article : Google Scholar : PubMed/NCBI
34	Tan BK, Adya R, Chen J, et al: Metformin decreases angiogenesis via NF-kappaB and Erk1/2/Erk5 pathways by increasing the antiangiogenic thrombospondin-1. Cardiovasc Res. 83:566–574. 2009. View Article : Google Scholar : PubMed/NCBI
35	González F, Nair KS, Daniels JK, Basal E and Schimke JM: Hyperandrogenism sensitizes mononuclear cells to promote glucose-induced inflammation in lean reproductive-age women. Am J Physiol Endocrinol Metab. 302:E297–E306. 2012. View Article : Google Scholar :