Effect of TNF-α on Molecules Related to the Insulin Action in Endometrial Cells Exposed to Hyperandrogenic and Hyperinsulinic Conditions Characteristics of Polycystic Ovary Syndrome

Abstract

Polycystic ovary syndrome (PCOS) affects not only ovarian functions but is also able to affect endometrium metabolism. Around 80% of women with PCOS are obese. High tumor necrosis factor (TNF)-α production and low adiponectin levels are characteristics of obesity. Interestingly, endometrium from obese women with PCOS presents an insulin-resistance condition, high TNF-α levels, and low adiponectin levels. However, TNF-α effect on molecules associated with insulin action in endometrial cells remains unclear. Therefore, the objective of this work was to evaluate TNF-α effect on expression of molecules associated with adiponectin (insulin sensitizing) and TNF-α signaling pathways and on Glucose Transporter type 4 (GLUT-4) levels in human endometrial cells under the characteristic conditions of hyperandrogenic/hyperinsulinic (HA/HI) PCOS. Two human endometrial stromal cell lines (T-HESC/St-T1b) under HA/HI conditions were used to assay the effect of high TNF-α concentration (100 ng/mL) on adiponectin, AdipoR1-AdipoR2 receptors, Adaptor protein phosphotyrosine interacting with PH domain and leucine zipper 1 (APPL1), Phospho-AMP-activated protein kinase T172 (p-AMPKT172), GLUT-4, Tumor necrosis factor receptor 1 (TNFR1)-Tumor necrosis factor receptor 2 (TNFR2) receptors protein levels, and nuclear factor κB (NFκB) nuclear content, by Western blot or immunocytochemistry. The NFκB participation in TNF-α effect on adiponectin expression was assayed using an NFκB inhibitor (pyrrolidine dithiocarbamate). The TNF-α increases the expression of molecules associated with its own signaling pathway (P < .05) and decreases the protein levels of adiponectin and its associated molecules (P < .05). Moreover, TNF-α increases NFκB nuclear content (P < .001), whereas with NFκB inhibition the decrease in adiponectin content induced by TNF-α was not observed. GLUT-4 levels were lower with TNF-α treatment (P < .01). Thus, in human endometrial stromal cells, high TNF-α levels negatively affect the insulin action through decreased adiponectin signaling and GLUT-4 protein. This could explain the failures observed in endometrial function of obese women with PCOS.

Keywords

endometrial cells TNF-α adiponectin insulin GLUT-4

Get full access to this article

View all access options for this article.

References

Azziz

Carmina

Dewailly

. Positions statement: criteria for defining polycystic ovary syndrome as a predominantly hyperandrogenic syndrome: an Androgen Excess Society guideline. J Clin Endocrinol Metab. 2006;91(11):4237–4245.

Azziz

Carmina

Chen

. Polycystic ovary syndrome. Nat Rev Dis Primers. 2016;2:16057.

Goodarzi

Dumesic

Chazenbalk

Azziz

. Polycystic ovary syndrome: etiology, pathogenesis and diagnosis. Nat Rev Endocrinol. 2011;7(4):219–231.

Orostica

Rosas

Plaza-Parrochia

. Altered steroid metabolism and insulin signaling in PCOS endometria: impact in tissue function. Curr Pharm Des. 2016;22(36):5614–5624.

Rosas

Gabler

Vantman

Romero

Vega

. Levels of rabs and WAVE family proteins associated with translocation of GLUT4 to the cell surface in endometria from hyperinsulinemic PCOS women. Hum Reprod. 2010;25(11):2870–2877.

Fornes

Ormazabal

Rosas

. Changes in the expression of insulin signaling pathway molecules in endometria from polycystic ovary syndrome women with or without hyperinsulinemia. Mol Med. 2010;16(3-4):129–136.

Carvajal

Rosas

Kohan

. Metformin augments the levels of molecules that regulate the expression of the insulin-dependent glucose transporter GLUT4 in the endometria of hyperinsulinemic PCOS patients. Hum Reprod. 2013;28(8):2235–2244.

Barber

McCarthy

Wass

Franks

. Obesity and polycystic ovary syndrome. Clin Endocrinol (Oxf). 2006;65(2):137–145.

Barber

Dimitriadis

Andreou

Franks

. Polycystic ovary syndrome: insight into pathogenesis and a common association with insulin resistance. Clin Med (Lond). 2016;16(3):262–266.

10.

Duleba

Dokras

. Is PCOS an inflammatory process? Fertil Steril. 2012;97(1):7–12.

11.

Garcia

Orostica

Poblete

Rosas

Astorga

Romero

. Endometria from obese PCOS women with hyperinsulinemia exhibit altered adiponectin signaling. Hor Metab Res. 2015;47(12):901–909.

12.

Oróstica

Astorga

Plaza-Parrochia

. Proinflammatory environment and role of TNF-alpha in endometrial function of obese women having polycystic ovarian syndrome. Int J Obes (Lond). 2016;40(11):1715–1722.

13.

Vilcek

Lee

. Tumor necrosis factor. New insights into the molecular mechanisms of its multiple actions. J Biol Chem. 1991;266(12):7313–7316.

14.

Hotamisligil

Shargill

Spiegelman

. Adipose expression of tumor necrosis factor-alpha: direct role in obesity-linked insulin resistance. Science. 1993;259(5091):87–91.

15.

Hotamisligil

Murray

Choy

Spiegelman

. Tumor necrosis factor alpha inhibits signaling from the insulin receptor. Proc Natl Acad Sci U S A. 1994;91(11):4854–4858.

16.

Hauner

Petruschke

Russ

Rohrig

Eckel

. Effects of tumour necrosis factor alpha (TNF alpha) on glucose transport and lipid metabolism of newly-differentiated human fat cells in cell culture. Diabetologia. 1995;38(7):764–771.

17.

Zhang

Gao

Yin

Quon

. S6K directly phosphorylates IRS-1 on Ser-270 to promote insulin resistance in response to TNF-(alpha) signaling through IKK2. J Biol Chem. 2008;283(51):35375–35382.

18.

Guerre-Millo

. Adiponectin: an update. Diabetes Metab. 2008;34(1):12–18.

19.

Wang

Jenkins

Trayhurn

. Expression and secretion of inflammation-related adipokines by human adipocytes differentiated in culture: integrated response to TNF-alpha. Am J Physiol. Endocrinol Metab. 2005;288(4):E731–E740.

20.

Wang

Trayhurn

. Acute and prolonged effects of TNF-alpha on the expression and secretion of inflammation-related adipokines by human adipocytes differentiated in culture. Pflugers Arch. 2006;452(4):418–427.

21.

Mirza

Shafique

Shaikh

Khan

Anwar Qureshi

. Association between circulating adiponectin levels and polycystic ovarian syndrome. J Ovarian Res. 2014;7:18.

22.

Krikun

Mor

Alvero

. A novel immortalized human endometrial stromal cell line with normal progestational response. Endocrinology. 2004;145(5):2291–2296.

23.

Zhang

. Pyrrolidine dithiocarbamate inhibits nuclear factor-kappaB pathway activation, and regulates adhesion, migration, invasion and apoptosis of endometriotic stromal cells. Mol Hum Reprod. 2011;17(3):175–181.

24.

Giudice

. Endometrium in PCOS: Implantation and predisposition to endocrine CA. Best Pract Res Clin Endocrinol Metab. 2006;20(2):235–244.

25.

Samalecos

Reimann

Wittmann

. Characterization of a novel telomerase-immortalized human endometrial stromal cell line, St-T1b. Reprod Biol Endocrinol. 2009;7:76.

26.

Ormazabal

Romero

Quest

Vega

. Testosterone modulates the expression of molecules linked to insulin action and glucose uptake in endometrial cells. Horm Metab Res. 2013;45(9):640–645.

27.

Quezada

Avellaira

Johnson

Gabler

Fuentes

Vega

. Evaluation of steroid receptors, coregulators, and molecules associated with uterine receptivity in secretory endometria from untreated women with polycystic ovary syndrome. Fertil Steril. 2006;85(4):1017–1026.

28.

Wajant

Pfizenmaier

Scheurich

. Tumor necrosis factor signaling. Cell Death Differ. 2003;10(1):45–65.

29.

Villavicencio

Goyeneche

Telleria

. Involvement of Akt, Ras and cell cycle regulators in the potential development of endometrial hyperplasia in women with polycystic ovarian syndrome. Gynecol Oncol. 2009;115(1):102–107.

30.

Plaza-Parrochia

Orostica

Garcia

. Molecular mechanisms of androstenediol in the regulation of the proliferative process of human endometrial cells. Reprod Sci. 2017;24(7):1079–1087.

31.

Bouwmeester

Bauch

Ruffner

. A physical and functional map of the human TNF-alpha/NF-kappa B signal transduction pathway. Nat Cell Biol. 2004;6(2):97–105.

32.

Fulghesu

Sanna

Uda

Magnini

Portoghese

Batetta

. 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.

33.

Vgontzas

Trakada

Bixler

. Plasma interleukin 6 levels are elevated in polycystic ovary syndrome independently of obesity or sleep apnea. Metabolism. 2006;55(8):1076–1082.

34.

Liu

. Transcriptional and post-translational regulation of adiponectin. Biochem J. 2009;425(1):41–52.

35.

Fasshauer

Kralisch

Klier

. Adiponectin gene expression and secretion is inhibited by interleukin-6 in 3T3-L1 adipocytes. Biochem Biophys Res Commun. 2003;301(4):1045–1050.

36.

Deepa

Dong

. APPL1: role in adiponectin signaling and beyond. Am J Physiol Endocrinol Metab. 2009;296(1):E22–E36.

37.

Mao

Kikani

Riojas

. APPL1 binds to adiponectin receptors and mediates adiponectin signalling and function. Nat Cell Biol. 2006;8(5):516–523.

38.

Wang

Xin

Xiang

. Yin-Yang regulation of adiponectin signaling by APPL isoforms in muscle cells. J Biol Chem. 2009;284(46):31608–31615.

39.

Cheng

Zhu

Chen

. The adaptor protein APPL2 inhibits insulin-stimulated glucose uptake by interacting with TBC1D1 in skeletal muscle. Diabetes. 2014;63(11):3748–3758.

40.

Patti

Kahn

. Nutrient sensor links obesity with diabetes risk. Nat Med. 2004;10(10):1049–1050.

41.

Rutledge

Adeli

. Fructose and the metabolic syndrome: pathophysiology and molecular mechanisms. Nutr Rev. 2007;65(6 pt 2):S13–S23.

42.

Kohan

Carvajal

Gabler

Vantman

Romero

Vega

. Role of the transcriptional factors FOXO1 and PPARG on gene expression of SLC2A4 in endometrial tissue from women with polycystic ovary syndrome. Reproduction. 2010;140(1):123–131.