Advanced glycation end products inhibit testosterone secretion by rat Leydig cells by inducing oxidative stress and endoplasmic reticulum stress

Zhao,Yun-Tao; Qi,Ya-Wei; Hu,Chuan-Yin; Chen,Shao-Hong; Liu,You

doi:10.3892/ijmm.2016.2645

Advanced glycation end products inhibit testosterone secretion by rat Leydig cells by inducing oxidative stress and endoplasmic reticulum stress

Authors:
- Yun-Tao Zhao
- Ya-Wei Qi
- Chuan-Yin Hu
- Shao-Hong Chen
- You Liu
View Affiliations

Affiliations: Modern Biochemistry Center, Guangdong Ocean University, Zhanjiang, Guangdong 524088, P.R. China, Institute of Plastic Surgery, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong 524001, P.R. China, Department of Biology, Guangdong Medical College, Zhanjiang, Guangdong 524023, P.R. China
Published online on: June 16, 2016 https://doi.org/10.3892/ijmm.2016.2645
Pages: 659-665

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

Diabetes severely impairs male reproduction. The present study assessed the effects and mechanisms of action of advanced glycation end products (AGEs), which play an important role in the development of diabetes complications, on testosterone secretion by rat Leydig cells. Primary rat Leydig cells were cultured and treated with AGEs (25, 50, 100 and 200 µg/ml). Testosterone production induced by human chorionic gonadotropin (hCG) was determined by ELISA. The mRNA and protein expression levels of steroidogenic acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (P450scc) and 3β-hydroxysteroid dehydrogenase (3β-HSD), which are involved in testosterone biosynthesis, were measured by reverse transcription-quantitative PCR and western blot analyssi, respectively. Reactive oxygen species (ROS) production in Leydig cells was measured using the dichlorofluorescein diacetate (DCFH-DA) probe. The expression levels of endoplasmic reticulum stress-related proteins [C/EBP homologous protein (CHOP) and glucose-regulated protein 78 (GRP78)] in the Leydig cells were measured by western blot analysis. We found that the AGEs markedly suppressed testosterone production by rat Leydig cells which was induced by hCG in a concentration-dependent manner compared with the control (P<0.01). The mRNA and protein expression levels of StAR, 3β-HSD and P450scc were downregulated by the AGEs in a dose-dependent manner compared with the control (P<0.01). The antioxidant agent, N-acetyl‑L‑cysteine (NAC), and the endoplasmic reticulum stress inhibitor, tauroursodeoxycholic acid (TUDCA), reversed the inhibitory effects of AGEs. In addition, the content of ROS in Leydig cells treated with AGEs increased significantly. The expression levels of CHOP and GRP78 were markedly upregulated by the AGEs in the Leydig cells. From these findings, it can be concluded that AGEs inhibit testosterone production by rat Leydig cells by inducing oxidative stress and endoplasmic reticulum stress.

View Figures

View References

1	Guariguata L, Whiting DR, Hambleton I, Beagley J, Linnenkamp U and Shaw JE: Global estimates of diabetes prevalence for 2013 and projections for 2035. Diabetes Res Clin Pract. 103:137–149. 2014. View Article : Google Scholar : PubMed/NCBI
2	Pinhas-Hamiel O, Dolan LM, Daniels SR, Standiford D, Khoury PR and Zeitler P: Increased incidence of non-insulin-dependent diabetes mellitus among adolescents. J Pediatr. 128:608–615. 1996. View Article : Google Scholar : PubMed/NCBI
3	Pinhas-Hamiel O and Zeitler P: The global spread of type 2 diabetes mellitus in children and adolescents. J Pediatr. 146:693–700. 2005. View Article : Google Scholar : PubMed/NCBI
4	Nadeau K and Dabelea D: Epidemiology of type 2 diabetes in children and adolescents. Endocr Res. 33:35–58. 2008. View Article : Google Scholar
5	Urakami T, Kubota S, Nitadori Y, Harada K, Owada M and Kitagawa T: Annual incidence and clinical characteristics of type 2 diabetes in children as detected by urine glucose screening in the Tokyo metropolitan area. Diabetes Care. 28:1876–1881. 2005. View Article : Google Scholar : PubMed/NCBI
6	Jangir RN and Jain GC: Diabetes mellitus induced impairment of male reproductive functions: a review. Curr Diabetes Rev. 10:147–157. 2014. View Article : Google Scholar : PubMed/NCBI
7	Sexton WJ and Jarow JP: Effect of diabetes mellitus upon male reproductive function. Urology. 49:508–513. 1997. View Article : Google Scholar : PubMed/NCBI
8	Baccetti B, La Marca A, Piomboni P, Capitani S, Bruni E, Petraglia F and De Leo V: Insulin-dependent diabetes in men is associated with hypothalamo-pituitary derangement and with impairment in semen quality. Hum Reprod. 17:2673–2677. 2002. View Article : Google Scholar : PubMed/NCBI
9	Schoeller EL, Schon S and Moley KH: The effects of type 1 diabetes on the hypothalamic, pituitary and testes axis. Cell Tissue Res. 349:839–847. 2012. View Article : Google Scholar : PubMed/NCBI
10	Scarano WR, Messias AG, Oliva SU, Klinefelter GR and Kempinas WG: Sexual behaviour, sperm quantity and quality after short-term streptozotocin-induced hyperglycaemia in rats. Int J Androl. 29:482–488. 2006. View Article : Google Scholar : PubMed/NCBI
11	Agbaje IM, Rogers DA, McVicar CM, McClure N, Atkinson AB, Mallidis C and Lewis SE: Insulin dependant diabetes mellitus: implications for male reproductive function. Hum Reprod. 22:1871–1877. 2007. View Article : Google Scholar : PubMed/NCBI
12	Ricci G, Catizone A, Esposito R, Pisanti FA, Vietri MT and Galdieri M: Diabetic rat testes: morphological and functional alterations. Andrologia. 41:361–368. 2009. View Article : Google Scholar : PubMed/NCBI
13	Mallidis C, Agbaje I, McClure N and Kliesch S: The influence of diabetes mellitus on male reproductive function: a poorly investigated aspect of male infertility. Urologe A. 50:33–37. 2011.In German. View Article : Google Scholar : PubMed/NCBI
14	La Vignera S, Condorelli R, Vicari E, D'Agata R and Calogero AE: Diabetes mellitus and sperm parameters. J Androl. 33:145–153. 2012. View Article : Google Scholar
15	Alves MG, Martins AD, Cavaco JE, Socorro S and Oliveira PF: Diabetes, insulin-mediated glucose metabolism and Sertoli/blood-testis barrier function. Tissue Barriers. 1:e239922013. View Article : Google Scholar
16	Vlassara H and Uribarri J: Advanced glycation end products (AGE) and diabetes: cause, effect, or both? Curr Diab Rep. 14:4532014. View Article : Google Scholar :
17	Vlassara H and Striker GE: Advanced glycation endproducts in diabetes and diabetic complications. Endocrinol Metab Clin North Am. 42:697–719. 2013. View Article : Google Scholar : PubMed/NCBI
18	Vlassara H and Striker GE: AGE restriction in diabetes mellitus: a paradigm shift. Nat Rev Endocrinol. 7:526–539. 2011. View Article : Google Scholar : PubMed/NCBI
19	Mallidis C, Agbaje I, Rogers D, Glenn J, McCullough S, Atkinson AB, Steger K, Stitt A and McClure N: Distribution of the receptor for advanced glycation end products in the human male reproductive tract: prevalence in men with diabetes mellitus. Hum Reprod. 22:2169–2177. 2007. View Article : Google Scholar : PubMed/NCBI
20	Moschonas DP, Piperi C, Korkolopoulou P, Levidou G, Kavantzas N, Trigka EA, Vlachos I, Arapostathi C, Perrea D, Mitropoulos D, et al: Impact of diet-induced obesity in male mouse reproductive system: the role of advanced glycation end product-receptor for advanced glycation end product axis. Exp Biol Med (Maywood). 239:937–947. 2014. View Article : Google Scholar
21	Mallidis C, Agbaje IM, Rogers DA, Glenn JV, Pringle R, Atkinson AB, Steger K, Stitt AW and McClure N: Advanced glycation end products accumulate in the reproductive tract of men with diabetes. Int J Androl. 32:295–305. 2009. View Article : Google Scholar
22	O'Neill J, Czerwiec A, Agbaje I, Glenn J, Stitt A, McClure N and Mallidis C: Differences in mouse models of diabetes mellitus in studies of male reproduction. Int J Androl. 33:709–716. 2010. View Article : Google Scholar
23	Mallidis C, Czerwiec A, Filippi S, O'Neill J, Maggi M and McClure N: Spermatogenic and sperm quality differences in an experimental model of metabolic syndrome and hypogonadal hypogonadism. Reproduction. 142:63–71. 2011. View Article : Google Scholar : PubMed/NCBI
24	Karimi J, Goodarzi MT, Tavilani H, Khodadadi I and Amiri I: Relationship between advanced glycation end products and increased lipid peroxidation in semen of diabetic men. Diabetes Res Clin Pract. 91:61–66. 2011. View Article : Google Scholar
25	Karimi J, Goodarzi MT, Tavilani H, Khodadadi I and Amiri I: Increased receptor for advanced glycation end products in spermatozoa of diabetic men and its association with sperm nuclear DNA fragmentation. Andrologia. 44(Suppl 1): 280–286. 2012. View Article : Google Scholar
26	Cheung KK, Luk AO, So WY, Ma RC, Kong AP, Chow FC and Chan JC: Testosterone level in men with type 2 diabetes mellitus and related metabolic effects: a review of current evidence. J Diabetes Investig. 6:112–123. 2015. View Article : Google Scholar : PubMed/NCBI
27	El Baba K and Azar ST: Low testosterone and diabetes. Curr Diabetes Rev. 9:418–421. 2013. View Article : Google Scholar : PubMed/NCBI
28	Iwashima Y, Eto M, Horiuchi S and Sano H: Advanced glycation end product-induced peroxisome proliferator-activated receptor gamma gene expression in the cultured mesangial cells. Biochem Biophys Res Commun. 264:441–448. 1999. View Article : Google Scholar : PubMed/NCBI
29	Chemes H, Cigorraga S, Bergadá C, Schteingart H, Rey R and Pellizzari E: Isolation of human Leydig cell mesenchymal precursors from patients with the androgen insensitivity syndrome: testosterone production and response to human chorionic gonadotropin stimulation in culture. Biol Reprod. 46:793–801. 1992. View Article : Google Scholar : PubMed/NCBI
30	Klinefelter GR, Hall PF and Ewing LL: Effect of luteinizing hormone deprivation in situ on steroidogenesis of rat Leydig cells purified by a multistep procedure. Biol Reprod. 36:769–783. 1987. View Article : Google Scholar : PubMed/NCBI
31	Bose HS, Lingappa VR and Miller WL: The steroidogenic acute regulatory protein, StAR, works only at the outer mitochondrial membrane. Endocr Res. 28:295–308. 2002. View Article : Google Scholar
32	Payne AH and Hales DB: Overview of steroidogenic enzymes in the pathway from cholesterol to active steroid hormones. Endocr Rev. 25:947–970. 2004. View Article : Google Scholar : PubMed/NCBI
33	Ott C, Jacobs K, Haucke E, Navarrete Santos A, Grune T and Simm A: Role of advanced glycation end products in cellular signaling. Redox Biol. 2:411–429. 2014. View Article : Google Scholar : PubMed/NCBI
34	Jahan H and Choudhary MI: Glycation, carbonyl stress and AGEs inhibitors: a patent review. Expert Opin Ther Pat. 25:1267–1284. 2015.PubMed/NCBI
35	Jin X, Yao T, Zhou Z, Zhu J, Zhang S, Hu W and Shen C: Advanced glycation end products enhance macrophages polarization into M1 phenotype through activating RAGE/NF-κB pathway. BioMed Res Int. 2015:7324502015. View Article : Google Scholar
36	Amaral S, Oliveira PJ and Ramalho-Santos J: Diabetes and the impairment of reproductive function: possible role of mitochondria and reactive oxygen species. Curr Diabetes Rev. 4:46–54. 2008. View Article : Google Scholar : PubMed/NCBI
37	Wautier MP, Chappey O, Corda S, Stern DM, Schmidt AM and Wautier JL: Activation of NADPH oxidase by AGE links oxidant stress to altered gene expression via RAGE. Am J Physiol Endocrinol Metab. 280:E685–E694. 2001.PubMed/NCBI
38	Rong G, Tang X, Guo T, Duan N, Wang Y, Yang L, Zhang J and Liang X: Advanced oxidation protein products induce apoptosis in podocytes through induction of endoplasmic reticulum stress. J Physiol Biochem. 71:455–470. 2015. View Article : Google Scholar : PubMed/NCBI
39	Xu J, Xiong M, Huang B and Chen H: Advanced glycation end products upregulate the endoplasmic reticulum stress in human periodontal ligament cells. J Periodontol. 86:440–447. 2015. View Article : Google Scholar
40	Wu L, Wang D, Xiao Y, Zhou X, Wang L, Chen B, Li Q, Guo X and Huang Q: Endoplasmic reticulum stress plays a role in the advanced glycation end product-induced inflammatory response in endothelial cells. Life Sci. 110:44–51. 2014. View Article : Google Scholar : PubMed/NCBI
41	Liu J, Huang K, Cai GY, Chen XM, Yang JR, Lin LR, Yang J, Huo BG, Zhan J and He YN: Receptor for advanced glycation end-products promotes premature senescence of proximal tubular epithelial cells via activation of endoplasmic reticulum stress-dependent p21 signaling. Cell Signal. 26:110–121. 2014. View Article : Google Scholar
42	Piperi C, Adamopoulos C, Dalagiorgou G, Diamanti-Kandarakis E and Papavassiliou AG: Crosstalk between advanced glycation and endoplasmic reticulum stress: emerging therapeutic targeting for metabolic diseases. J Clin Endocrinol Metab. 97:2231–2242. 2012. View Article : Google Scholar : PubMed/NCBI