Skip to main content
Log in

Therapeutic prospects of hydroxytyrosol on experimentally induced diabetic testicular damage: potential interplay with AMPK expression

  • Regular Article
  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Abstract

Male reproductive dysfunction represents one of the overlooked consequences of diabetes that still deserve more scientific attention. We designed this study to explore the therapeutic potential of hydroxytyrosol (HT) on diabetic testicular damage and to investigate its relationship with adenosine monophosphate-activated protein kinase (AMPK) expression. In this context, 30 adult male Wistar rats were utilized and subdivided into control, diabetic and HT-treated diabetic groups. Testicular sections were prepared for histopathological examination and immunohistochemical detection of 8-hydroxy-2′-deoxyguanosine, Sertoli cell vimentin, myoid cell α-SMA, androgen receptors and caspase-3. We also assessed oxidative enzymatic and lipid peroxidation biochemical profiles, sperm count, morphology and motility. Real-time PCR of AMPK expression in tissue homogenate was performed. We observed that HT restored testicular histopathological structure and significantly reduced oxidative DNA damage and the apoptotic index. The HT-treated group also exhibited significantly higher Sertoli cell vimentin, myoid cell α-SMA and androgen receptor immune expression than the diabetic group. A rescue of the oxidative enzymatic activity, lipid peroxidation profiles, sperm count, morphology and motility to control levels was also evident in the HT-treated group. Significant upregulation of AMPK mRNA expression in the HT-treated group clarified the role of AMPK as an underlying molecular interface of the ameliorative effects of HT. We concluded that HT exhibited tangible antioxidant and antiapoptotic impacts on the testicular cytomorphological and immunohistochemical effects of experimentally induced diabetes. Furthermore, AMPK has an impactful role in the molecular machinery of these effects.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2.
Fig 3
Fig. 4
Fig. 5.
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11
Fig. 12
Fig. 13.

Similar content being viewed by others

Abbreviations

HT:

hydroxytyrosol

AMPK:

adenosine mono-phosphate-activated protein kinase

α-SMA:

α smooth muscle actin

References

  • Abdelrahman SA, Samak MA, Shalaby SM (2018) Fluoxetine pretreatment enhances neurogenic, angiogenic and immunomodulatory effects of MSCs on experimentally induced diabetic neuropathy. Cell Tissue Res 374:83–97

    Article  CAS  PubMed  Google Scholar 

  • Agbaje I, Rogers DA, McVicar CM, McClure N, Atkinson AB, Mallidis C, Lewis SEM (2007) Insulin dependant diabetes mellitus: implications for male reproductive function. Hum Reprod 22(7):1871–1877

    Article  CAS  PubMed  Google Scholar 

  • Aghanoori MR, Smith DR, Shariati-Ievari S, Ajisebutu A, Nguyen A, Desmond F, Fernyhough P (2019) Insulin-like growth factor-1 activates AMPK to augment mitochondrial function and correct neuronal metabolism in sensory neurons in type 1 diabetes. Mol Metab 20:149–165

    Article  CAS  PubMed  Google Scholar 

  • Alves MG, Martins AD, Cavaco JE, Socorro S, Oliveira PF (2013) Diabetes, insulin-mediated glucose metabolism and Sertoli/blood-testis barrier function. Tissue Barriers 1(2):e23992

    Article  PubMed  PubMed Central  Google Scholar 

  • Avola R, Graziano ACE, Pannuzzo G, Bonina F, Cardile V (2019) Hydroxytyrosol from olive fruits prevents blue-light-induced damage in human keratinocytes and fibroblasts. J Cell Physiol 234(6):9065–9076

    Article  CAS  PubMed  Google Scholar 

  • Bernardino RL, Alves MG., & Oliveira PF (2018) Evaluation of the purity of sertoli cell primary cultures. In: Sertoli Cells. Humana Press, New York, pp 9–15

    Chapter  Google Scholar 

  • Bertoldo M, Faure M, Dupont J, Froment P (2015) AMPK: a master energy regulator for gonadal function. Front Neurosci 9:235

    Article  PubMed  PubMed Central  Google Scholar 

  • Busetto GM, Agarwal A, Virmani A, Antonini G, Ragonesi G, Del Giudice F, De Berardinis E (2018) Effect of metabolic and antioxidant supplementation on sperm parameters in oligo-astheno-teratozoospermia, with and without varicocele: A double-blind placebo-controlled study. Andrologia 50(3):e12927

    Article  CAS  Google Scholar 

  • Calabriso, N., Gnoni, A., Stanca, E., Cavallo, A., Damiano, F., Siculella, L., & Carluccio, MA. (2018). Hydroxytyrosol ameliorates endothelial function under inflammatory conditions by preventing mitochondrial dysfunction. Oxid Med Cell Longev

  • Chen LY, Brown PR, Willis WB, Eddy EM (2014) Peritubularmyoid cells participate in male mouse spermatogonial stem cell maintenance. Endocrinology 155(12):4964–4974

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Colica, C., Di Renzo, L., Trombetta, D., Smeriglio, A., Bernardini, S., Cioccoloni, G., & De Lorenzo, A. (2017). Antioxidant effects of a hydroxytyrosolbased pharmaceutical formulation on body composition, metabolic state, and gene expression: a randomized double-blinded, placebo-controlled crossover trial. Oxid Med Cell Longev

  • Condorelli RA, La Vignera S, Mongioì LM, Alamo A, Calogero AE (2018) Diabetes mellitus and infertility: different pathophysiological effects in type 1 and type 2 on sperm function. Front Endocrinol 9:268

    Article  Google Scholar 

  • de las López Hazas MC, Godinho-Pereira J, Macià A, Almeida AF, Ventura MR, Motilva MJ, Santos CN (2018) Brain uptake of hydroxytyrosol and its main circulating metabolites: Protective potential in neuronal cells. J Funct Foods 46:110–117

    Article  CAS  Google Scholar 

  • Echeverría F, Ortiz M, Valenzuela R, Videla L (2017) Hydroxytyrosol and cytoprotection: a projection for clinical interventions. Int J Mol Sci 18(5):930

    Article  PubMed Central  CAS  Google Scholar 

  • El-azem, N., Pulido-Moran, M., Ramirez-Tortosa, CL., Quiles, JL., Cara, FE., Sanchez-Rovira, P., & Ramirez-Tortosa, M. (2019). Modulation by hydroxytyrosol of oxidative stress and antitumor activities of paclitaxel in breast cancer. Eur J Nutr, 58(3):1203–1211

  • Elewa, YH., Mohamed, AA. R., Galal, AA., El-naseery, NI., Ichii, O., & Kon, Y. (2019). Food Yellow4 reprotoxicity in relation to localization of DMC1 and apoptosis in rat testes: Roles of royal jelly and cod liver oil. Ecotoxicology and environmental safety, 169:696–706

  • Erkekoglu P, Zeybek ND, Giray B, Asan E, Hincal F (2012) The effects of di (2-ethylhexyl) phthalate exposure and selenium nutrition on sertoli cell vimentin structure and germ-cell apoptosis in rat testis. Arch Environ Contam Toxicol 62(3):539–547

    Article  CAS  PubMed  Google Scholar 

  • Fuccelli R, Fabiani R, Rosignoli P (2018) Hydroxytyrosol exerts anti-inflammatory and anti-oxidant activities in a mouse model of systemic inflammation. Molecules 23(12):3212

    Article  PubMed Central  CAS  Google Scholar 

  • Funakohi-Tago M, Sakata T, Fujiwara S, Sakakura A, Sugai T, Tago K, Tamura H (2018) Hydroxytyrosol butyrate inhibits 6-OHDA-induced apoptosis through activation of the Nrf2/HO-1 axis in SH-SY5Y cells. Eur J Pharmacol 834:246–256

    Article  CAS  PubMed  Google Scholar 

  • Griswold MD (2018) 50 years of spermatogenesis: Sertoli cells and their interactions with germ cells. Biol Reprod 99(1):87–100

    Article  PubMed  PubMed Central  Google Scholar 

  • Hawley SA, Pan DA, Mustard KJ, Ross L, Bain J, Edelman AM, Hardie DG (2005) Calmodulin-dependent protein kinase kinase-β is an alternative upstream kinase for AMP-activated protein kinase. Cell Metab 2(1):9–19

    Article  CAS  PubMed  Google Scholar 

  • Institutional Animal Care and Use Committee (IACUC), Office of Research Compliance (ORC) (2013) https://resea rch.iu.edu/doc/compliance/animal-care/bloomin ton/iub-biacuc-non-pharmaceutical-and-pharmaceutical-grade-compounds-in-researchanimals.pdf

  • Jiang X, Bai Y, Zhang Z, Xin Y, Cai L (2014) Protection by sulforaphane from type 1 diabetes-induced testicular apoptosis is associated with the up-regulation of Nrf2 expression and function. Toxicol Appl Pharmacol 279(2):198–210

    Article  CAS  PubMed  Google Scholar 

  • Jiao D, Zhang H, Jiang Z, Huang W, Liu Z, Wang Z, Wu H (2018) MicroRNA-34a targets sirtuin 1 and leads to diabetes-induced testicular apoptotic cell death. J Mol Med 96(9):939–949

    Article  CAS  PubMed  Google Scholar 

  • Johnston DS, Russell LD, Friel PJ, Griswold MD (2001) Murine germ cells do not require functional androgen receptors to complete spermatogenesis following spermatogonial stem cell transplantation. Endocrinology 142(6):2405–2408

    Article  CAS  PubMed  Google Scholar 

  • Joseph BK, Liu HY, Francisco J, Pandya D, Donigan M, Gallo-Ebert C, Nickels JT (2015) Inhibition of AMP kinase by the protein phosphatase 2A heterotrimer, PP2APpp2r2d. J Biol Chem 290(17):10588–10598

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Karakas M, Schäfer S, Appelbaum S, Ojeda F, Kuulasmaa K, Brückmann B, Palosaari T (2018) Testosterone levels and type 2 diabetes—no correlation with age, differential predictive value in men and women. Biomolecules 8(3):76

    Article  PubMed Central  CAS  Google Scholar 

  • Kaur G, Thompson LA, Dufour JM (2014) Sertoli cells–immunological sentinels of spermatogenesis. In: Seminars in Cell & Developmental Biology, vol 30. Academic Press, pp 36–44

  • Kedechi S, Zribi N, Louati N, Menif H, Sellami A, Lassoued S, Chakroun N (2017) Antioxidant effect of hydroxytyrosol on human sperm quality during in vitro incubation. Andrologia 49(1):e12595

    Article  CAS  Google Scholar 

  • Khaneshi F, Nasrolahi O, Azizi S, Nejati V (2013) Sesame effects on testicular damage in streptozotocin-induced diabetes rats. Avicenna J Phytomed 3(4):347

    PubMed  PubMed Central  Google Scholar 

  • Kilarkaje N, Al-Hussaini H, Al-Bader MM (2014) Diabetes-induced DNA damage and apoptosis are associated with poly (ADP ribose) polymerase 1 inhibition in the rat testis. Eur J Pharmacol 737:29–40

    Article  CAS  PubMed  Google Scholar 

  • Kushwaha S, Jena GB (2013) Telmisartan ameliorates germ cell toxicity in the STZ-induced diabetic rat: studies on possible molecular mechanisms. Mutat Res Genet Toxicol Environ Mutagen 755(1):11–23

    Article  CAS  Google Scholar 

  • Kwan HY, Chao X, Su T, Fu X, Tse AKW, Fong WF, Yu ZL (2017) The anticancer and antiobesity effects of Mediterranean diet. Crit Rev Food Sci Nutr 57(1):82–94

    Article  CAS  PubMed  Google Scholar 

  • La Vignera S, Calogero AE, Condorelli R, Lanzafame F, Giammusso B, Vicari E (2009) Andrological characterization of the patient with diabetes mellitus. Minerva Endocrinol 34(1):1–9

    PubMed  Google Scholar 

  • Lert Amornpat T, Maketon C, Fungfuang W (2017) Effect of Kaempferiaparviflora on sexual performance in streptozotocin-induced diabetic male rats. Andrologia 49(10):1–6

    Article  CAS  Google Scholar 

  • Miao J, Huang Z, Liu S, Li X, Jia P, Guo Y, Jia D (2019) Hydroxytyrosol protects against myocardial ischemia reperfusion injury by inhibiting mitochondrial permeability transition pore opening. Exp Ther Med 17(1):671–678

    CAS  PubMed  Google Scholar 

  • Pérez CV, Theas MS, Jacobo PV, Jarazo-Dietrich S, Guazzone VA, Lustig L (2013) Dual role of immune cells in the testis: protective or pathogenic for germ cells? Spermatogenesis 3(1):e23870

    Article  PubMed  PubMed Central  Google Scholar 

  • Prabhu A, Xu Q, Manigrasso MB, Biswas M, Flynn E, Iliescu R, Maric C (2010) Expression of aromatase, androgen and estrogen receptors in peripheral target tissues in diabetes. Steroids 75(11):779–787

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rato L, Alves MG, Duarte AI, Santos MS, Moreira PI, Cavaco JE, Oliveira PF (2015) Testosterone deficiency induced by progressive stages of diabetes mellitus impairs glucose metabolism and favors glycogenesis in mature rat Sertoli cells. Int J Biochem Cell Biol 66:1–10

    Article  CAS  PubMed  Google Scholar 

  • Rato L, Oliveira PF, Sousa M, Silva BM, Alves MG (2019) Role of reactive oxygen species in diabetes-induced male reproductive dysfunction. In: Oxidants, antioxidants and impact of the oxidative status in male reproduction. Academic Press, pp 135–147

  • Rebourcet D, O’Shaughnessy PJ, Monteiro A, Milne L, Cruickshanks L, Jeffrey N, Smith LB (2014) Sertoli cells maintain Leydig cell number and peritubularmyoid cell activity in the adult mouse testis. PloS One 9(8):e105687

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Ricci G, Catizone A, Esposito R, Pisanti FA, Vietri MT, Galdieri M (2009) Diabetic rat testes: morphological and functional alterations. Andrologia 41(6):361–368

    Article  CAS  PubMed  Google Scholar 

  • Rodríguez-Morató J, Boronat A, Kotronoulas A, Pujadas M, Pastor A, Olesti E, de la Torre R (2016) Metabolic disposition and biological significance of simple phenols of dietary origin: hydroxytyrosol and tyrosol. Drug Metab Rev 48(2):218–236

    Article  PubMed  CAS  Google Scholar 

  • Schoeller EL, Albanna G, Frolova AI, Moley KH (2012) Insulin rescues impaired spermatogenesis via the hypothalamic-pituitary-gonadal axis in Akita diabetic mice and restores male fertility. Diabetes 61(7):1869–1878

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Soliman GA, Saeedan AS, Abdel-Rahman RF, Ogaly HA, Abd-Elsalam RM, Abdel-Kader MS (2019) Olive leaves extract attenuates type II diabetes mellitus-induced testicular damage in rats: molecular and biochemical study. Saudi Pharm J 27(3):326–340

    Article  PubMed  Google Scholar 

  • Sönmez MF, Kılıç E, Karabulut D, Çilenk K, Deligönül E, Dündar M (2016) Nitric oxide synthase in diabetic rat testicular tissue and the effects of pentoxifylline therapy. Syst Biol Reprod Med 62(1):22–30

    Article  PubMed  CAS  Google Scholar 

  • Tartarin P, Guibert E, Touré A, Ouiste C, Leclerc J, Sanz N, McNeilly AS (2012) Inactivation of AMPKα1 induces asthenozoospermia and alters spermatozoa morphology. Endocrinology 153(7):3468–3481

    Article  CAS  PubMed  Google Scholar 

  • Toyokuni S, Tanaka T, Hattori Y, Nishiyama Y, Yoshida A (1997) Quantitative immunohistochemical determination of 8-hydroxy-2'-deoxyguanosine by a monoclonal antibody N45. 1: its application to ferric nitrilotriacetate-induced renal carcinogenesis model. Lab Invest 3:365374

    Google Scholar 

  • Tuck KL, Hayball PJ (2002) Major phenolic compounds in olive oil: metabolism and health effects. J Nutr Biochem 13(11):636–644

    Article  CAS  PubMed  Google Scholar 

  • Valenzuela R, Illesca P, Echeverría F, Espinosa A, Rincón-Cervera MÁ, Ortiz M, Videla LA (2017) Molecular adaptations underlying the beneficial effects of hydroxytyrosol in the pathogenic alterations induced by a high-fat diet in mouse liver: PPAR-α and Nrf2 activation, and NF-κB down-regulation. Food Funct 8(4):1526–1537

    Article  CAS  PubMed  Google Scholar 

  • Valls-Pedret C, Sala-Vila A, Serra-Mir M, Corella D, De la Torre R, Martínez-González MÁ, Estruch R (2015) Mediterranean diet and age-related cognitive decline: a randomized clinical trial. JAMA Intern Med 175(7):1094–1103

    Article  PubMed  Google Scholar 

  • Vorilhon S, Brugnon F, Kocer A, Dollet S, Bourgne C, Berger M, Gharagozloo P (2018) Accuracy of human sperm DNA oxidation quantification and threshold determination using an 8-OHdG immuno-detection assay. Hum Reprod 33(4):553–562

    Article  CAS  PubMed  Google Scholar 

  • Wang N, Zhang J, Qin M, Yi W, Yu S, Chen Y, Zhang R (2018) Amelioration of streptozotocin-induced pancreatic β cell damage by morin: Involvement of the AMPK-FOXO3-catalase signaling pathway. Int J Mol Med 41(3):1409–1418

    CAS  PubMed  Google Scholar 

  • Wild S, Roglic G, Green A, Sicree R, King H (2004) Global prevalence of diabetes: estimates for the year 2000 and projections for 2030. Diabetes Care 27(5):1047–1053

    Article  PubMed  Google Scholar 

  • Wu LX, Xu YY, Yang ZJ, Feng Q (2018a) Hydroxytyrosol and olive leaf extract exert cardioprotective effects by inhibiting GRP78 and CHOP expression. J Biomed Res 32(5):371

    PubMed  PubMed Central  Google Scholar 

  • Wu X, Luo P, Rao W, Dai S, Zhang L, Ma W, Fei Z (2018b) Homer1a attenuates hydrogen peroxide-induced oxidative damage in HT-22 cells through AMPK-dependent autophagy. Front Neurosci 12:51

    Article  PubMed  PubMed Central  Google Scholar 

  • Yamada M, Kurihara H, Kinoshita K, Sakai T (2005) Temporal expression of alpha–smooth muscle actin and drebrin in septal interstitial cells during alveolar maturation. J Histochem Cytochem 53(6):735–744

    Article  CAS  PubMed  Google Scholar 

  • Zawada I, Masternak MM, List EO, Stout MB, Berryman DE, Lewinski A, Gesing A (2015) Gene expression of key regulators of mitochondrial biogenesis is sex dependent in mice with growth hormone receptor deletion in liver. Aging (Albany NY) 7(3):195

    Article  CAS  Google Scholar 

  • Zheng A, Li H, Xu J, Cao K, Li H, Pu W, Feng Z (2015) Hydroxytyrosol improves mitochondrial function and reduces oxidative stress in the brain of db/db mice: role of AMP-activated protein kinase activation. Br J Nutr 113(11):1667–1676

    Article  CAS  PubMed  Google Scholar 

  • Zhong RZ, Zhou DW (2013) Oxidative stress and role of natural plant derived antioxidants in animal reproduction. J Integr Agric 12(10):1826–1838

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Amira E. Alsemeh.

Ethics declarations

Conflict of interest

The authors declared that they have no conflicts of interest.

Ethical approval

All procedures were performed in accordance to guidelines of IACUC (Institutional Animal Care and Use Committee), Zagazig University, Egypt.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Alsemeh, A.E., Samak, M.A. & El-Fatah, S.S.A. Therapeutic prospects of hydroxytyrosol on experimentally induced diabetic testicular damage: potential interplay with AMPK expression. Cell Tissue Res 380, 173–189 (2020). https://doi.org/10.1007/s00441-019-03143-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00441-019-03143-2

Keywords

Navigation