Protective effects of physiological testosterone on advanced glycation end product‑induced injury in human endothelial cells

Xie,Yaping; Yu,Dan; Wu,Jiahua; Li,Lin

doi:10.3892/mmr.2017.6130

Protective effects of physiological testosterone on advanced glycation end product‑induced injury in human endothelial cells

Authors:
- Yaping Xie
- Dan Yu
- Jiahua Wu
- Lin Li
View Affiliations

Affiliations: Department of Hematology, Hangzhou No. 1 People's Hospital, Hangzhou, Zhejiang 310016, P.R. China, Department of Endocrinology, The Affiliated Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310016, P.R. China
Published online on: January 19, 2017 https://doi.org/10.3892/mmr.2017.6130
Pages: 1165-1171
Copyright: © Xie et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The effect of testosterone, a sex steroid, on endothelial cells is controversial as it is uncertain if it has a protective effect on them. Whether physiological testosterone can inhibit the deleterious effects of advanced glycation end products (AGEs) on endothelial cells remains to be elucidated. The present study focused on elucidating the effect of testosterone on the injury of endothelial cells induced by AGEs. Human umbilical vein endothelial cells (HUVECs) were cultured in vitro and treated with AGEs in the presence or absence of various concentrations of testosterone. The cell viability in each group was measured using an MTS assay. Early‑stage apoptosis was detected using flow cytometry with Annexin V‑fluorescein isothiocyanate/propidium iodide double staining, and the expression levels of apoptosis‑associated proteins, B cell lymphoma‑2 (Bcl‑2), Bcl‑2‑associated X protein (Bax) and caspase‑3, were determined using western blot analysis. Oxidative stress and pro‑inflammatory parameters in the medium were evaluated using an enzyme‑linked immunosorbent assay. The MTS results showed that AGEs significantly decreased the proliferation of HUVECs, whereas a physiological concentration of testosterone alleviated this damage. Physiological concentrations of testosterone protected the HUVECs from AGE‑induced apoptosis, mediated by caspase‑3 and Bax/Bcl‑2. In addition, treatment of the HUVECs with AGEs caused a significant decrease in anti‑oxidative parameters, but increased the concentrations of malondialdehyde and tumor necrosis factor‑α. The activation of Janus kinase 2 and signal transducer and activator of transcription 3 was significantly increased by incubation with AGEs. However, pre‑incubation with a physiological concentration of testosterone attenuated these changes. Therefore, the data obtained in the present study established the potential role of physiological testosterone in ameliorating AGE‑induced damage in HUVECs.

View Figures

View References

1	Rahman S, Rahman T, Ismail AA and Rashid AR: Diabetes-associated macrovasculopathy: Pathophysiology and pathogenesis. Diabetes Obes Metab. 9:767–780. 2007. View Article : Google Scholar : PubMed/NCBI
2	Cooper ME, Bonnet F, Oldfield M and Jandeleit-Dahm K: Mechanisms of diabetic vasculopathy: An overview. Am J Hypertens. 14:475–486. 2001. View Article : Google Scholar : PubMed/NCBI
3	Neves D: Advanced glycation end-products: A common pathway in diabetes and age-related erectile dysfunction. Free Radic Res 47 Suppl. 1:49–69. 2013. View Article : Google Scholar
4	Prasad A, Bekker P and Tsimikas S: Advanced glycation end products and diabetic cardiovascular disease. Cardiol Rev. 20:177–183. 2012. View Article : Google Scholar : PubMed/NCBI
5	Basta G, Schmidt AM and De Caterina R: Advanced glycation end products and vascular inflammation: Implications for accelerated atherosclerosis in diabetes. Cardiovasc Res. 63:582–592. 2004. View Article : Google Scholar : PubMed/NCBI
6	Goldin A, Beckman JA, Schmidt AM and Creager MA: Advanced glycation end products: Sparking the development of diabetic vascular injury. Circulation. 114:597–605. 2006. View Article : Google Scholar : PubMed/NCBI
7	Min C, Kang E, Yu SH, Shinn SH and Kim YS: Advanced glycation end products induce apoptosis and procoagulant activity in cultured human umbilical vein endothelial cells. Diabetes Res Clin Pract. 46:197–202. 1999. View Article : Google Scholar : PubMed/NCBI
8	Mendelsohn ME and Karas RH: The protective effects of estrogen on the cardiovascular system. N Engl J Med. 340:1801–1811. 1999. View Article : Google Scholar : PubMed/NCBI
9	Bagatell CJ and Bremner WJ: Androgens in men-uses and abuses. N Engl J Med. 334:707–714. 1996. View Article : Google Scholar : PubMed/NCBI
10	Sullivan ML, Martinez CM, Gennis P and Gallagher EJ: The cardiac toxicity of anabolic steroids. Prog Cardiovasc Dis. 41:1–15. 1998. View Article : Google Scholar : PubMed/NCBI
11	Jones RD, Nettleship JE, Kapoor D, Jones HT and Channer KS: Testosterone and atherosclerosis in aging men: Purported association and clinical implications. Am J Cardiovasc Drugs. 5:141–154. 2005. View Article : Google Scholar : PubMed/NCBI
12	Choi BG and McLaughlin MA: Why men's hearts break: Cardiovascular effects of sex steroids. Endocrinol Metab Clin North Am. 36:365–377. 2007. View Article : Google Scholar : PubMed/NCBI
13	Smith AM, English KM, Malkin CJ, Jones RD, Jones TH and Channer KS: Testosterone does not adversely affect fibrinogen or tissue plasminogen activator (tPA) and plasminogen activator inhibitor-1 (PAI-1) levels in 46 men with chronic stable angina. Eur J Endocrinol. 152:285–291. 2005. View Article : Google Scholar : PubMed/NCBI
14	Adams MR, Williams JK and Kaplan JR: Effects of androgens on coronary artery atherosclerosis and atherosclerosis-related impairment of vascular responsiveness. Arterioscler Thromb Vasc Biol. 15:562–570. 1995. View Article : Google Scholar : PubMed/NCBI
15	Nathan L, Shi W, Dinh H, Mukherjee TK, Wang X, Lusis AJ and Chaudhuri G: Testosterone inhibits early atherogenesis by conversion to estradiol: Critical role of aromatase. Proc Natl Acad Sci USA. 98:3589–3593. 2001. View Article : Google Scholar : PubMed/NCBI
16	English KM, Mandour O, Steeds RP, Diver MJ, Jones TH and Channer KS: Men with coronary artery disease have lower levels of androgens than men with normal coronary angiograms. Eur Heart J. 21:890–894. 2000. View Article : Google Scholar : PubMed/NCBI
17	Jin H, Lin J, Fu L, Mei YF, Peng G, Tan X, Wang DM, Wang W and Li YG: Physiological testosterone stimulates tissue plasminogen activator and tissue factor pathway inhibitor and inhibits plasminogen activator inhibitor type 1 release in endothelial cells. Biochem Cell Biol. 85:246–251. 2007. View Article : Google Scholar : PubMed/NCBI
18	Jin H, Wang DY, Mei YF, Qiu WB, Zhou Y, Wang DM, Tan XR and Li YG: Mitogen-activated protein kinases pathway is involved in physiological testosterone-induced tissue factor pathway inhibitor expression in endothelial cells. Blood Coagul Fibrinolysis. 21:420–424. 2010. View Article : Google Scholar : PubMed/NCBI
19	Cai J, Hong Y, Weng C, Tan C, Imperato-McGinley J and Zhu YS: Androgen stimulates endothelial cell proliferation via an androgen receptor/VEGF/cyclin A-mediated mechanism. Am J Physiol Heart Circ Physiol. 300:H1210–H1221. 2011. View Article : Google Scholar : PubMed/NCBI
20	Yu J, Akishita M, Eto M, Ogawa S, Son BK, Kato S, Ouchi Y and Okabe T: Androgen receptor-dependent activation of endothelial nitric oxide synthase in vascular endothelial cells: Role of phosphatidylinositol 3-kinase/akt pathway. Endocrinology. 151:1822–1828. 2010. View Article : Google Scholar : PubMed/NCBI
21	Xu Y, Feng L, Wang S, Zhu Q, Zheng Z, Xiang P, He B and Tang D: Calycosin protects HUVECs from advanced glycation end products-induced macrophage infiltration. J Ethnopharmacol. 137:359–370. 2011. View Article : Google Scholar : PubMed/NCBI
22	Zhuang X, Pang X, Zhang W, Wu W, Zhao J, Yang H and Qu W: Effects of zinc and manganese on advanced glycation end products (AGEs) formation and AGEs-mediated endothelial cell dysfunction. Life Sci. 90:131–139. 2012. View Article : Google Scholar : PubMed/NCBI
23	Rashid G, Benchetrit S, Fishman D and Bernheim J: Effect of advanced glycation end-products on gene expression and synthesis of TNF-alpha and endothelial nitric oxide synthase by endothelial cells. Kidney Int. 66:1099–1106. 2004. View Article : Google Scholar : PubMed/NCBI
24	Grimm S, Ott C, Horlacher M, Weber D, Hohn A and Grune T: Advanced-glycation-end-product-induced formation of immunoproteasomes: Involvement of RAGE and Jak2/STAT1. Biochem J. 448:127–139. 2012. View Article : Google Scholar : PubMed/NCBI
25	Nedić O, Rattan SI, Grune T and Trougakos IP: Molecular effects of advanced glycation end products on cell signalling pathways, ageing and pathophysiology. Free Radic Res. 47:(Suppl 1). S28–S38. 2013. View Article : Google Scholar
26	Rodiño-Janeiro BK, González-Peteiro M, Ucieda-Somoza R, González-Juanatey JR and Alvarez E: Glycated albumin, a precursor of advanced glycation end-products, up-regulates NADPH oxidase and enhances oxidative stress in human endothelial cells: Molecular correlate of diabetic vasculopathy. Diabetes Metab Res Rev. 26:550–558. 2010. View Article : Google Scholar : PubMed/NCBI
27	Orasanu G and Plutzky J: The pathologic continuum of diabetic vascular disease. J Am Coll Cardiol. 53:(Suppl 5). S35–S42. 2009. View Article : Google Scholar : PubMed/NCBI
28	Méndez JD, Xie J, Aguilar-Hernández M and Méndez-Valenzuela V: Trends in advanced glycation end products research in diabetes mellitus and its complications. Mol Cell Biochem. 341:33–41. 2010. View Article : Google Scholar : PubMed/NCBI
29	Sang HQ, Gu JF, Yuan JR, Zhang MH, Jia XB and Feng L: The protective effect of Smilax glabra extract on advanced glycation end products-induced endothelial dysfunction in HUVECs via RAGE-ERK1/2-NF-κB pathway. J Ethnopharmacol. 155:785–795. 2014. View Article : Google Scholar : PubMed/NCBI
30	Alexandersen P, Haarbo J, Byrjalsen I, Lawaetz H and Christiansen C: Natural androgens inhibit male atherosclerosis: A study in castrated, cholesterol-fed rabbits. Circ Res. 84:813–819. 1999. View Article : Google Scholar : PubMed/NCBI
31	Nettleship JE, Jones TH, Channer KS and Jones RD: Physiological testosterone replacement therapy attenuates fatty streak formation and improves high-density lipoprotein cholesterol in the Tfm mouse: An effect that is independent of the classic androgen receptor. Circulation. 116:2427–2434. 2007. View Article : Google Scholar : PubMed/NCBI
32	Hulsmans M and Holvoet P: The vicious circle between oxidative stress and inflammation in atherosclerosis. J Cell Mol Med. 14:70–78. 2010. View Article : Google Scholar : PubMed/NCBI
33	Ho E, Galougahi Karimi K, Liu CC, Bhindi R and Figtree GA: Biological markers of oxidative stress: Applications to cardiovascular research and practice. Redox Biol. 1:483–491. 2013. View Article : Google Scholar : PubMed/NCBI
34	Allen JD, Giordano T and Kevil CG: Nitrite and nitric oxide metabolism in peripheral artery disease. Nitric Oxide. 26:217–222. 2012. View Article : Google Scholar : PubMed/NCBI
35	Huang JS, Guh JY, Hung WC, Yang ML, Lai YH, Chen HC and Chuang LY: Role of the Janus kinase (JAK)/signal transducters and activators of transcription (STAT) cascade in advanced glycation end-product-induced cellular mitogenesis in NRK-49F cells. Biochem J. 342:231–238. 1999. View Article : Google Scholar : PubMed/NCBI