Volume 41 Issue 5
Sep.  2020
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Rong-Nan Li, Zhen-Dong Zhu, Yi Zheng, Ying-Hua Lv, Xiu-E Tian, De Wu, Yong-Jun Wang, Wen-Xian Zeng. Metformin improves boar sperm quality via 5′-AMP-activated protein kinase-mediated energy metabolism in vitro. Zoological Research, 2020, 41(5): 527-538. doi: 10.24272/j.issn.2095-8137.2020.074
Citation: Rong-Nan Li, Zhen-Dong Zhu, Yi Zheng, Ying-Hua Lv, Xiu-E Tian, De Wu, Yong-Jun Wang, Wen-Xian Zeng. Metformin improves boar sperm quality via 5′-AMP-activated protein kinase-mediated energy metabolism in vitro. Zoological Research, 2020, 41(5): 527-538. doi: 10.24272/j.issn.2095-8137.2020.074

Metformin improves boar sperm quality via 5′-AMP-activated protein kinase-mediated energy metabolism in vitro

doi: 10.24272/j.issn.2095-8137.2020.074
Funds:  This study was supported in part by the National Key R&D Program of China (2018YFD0501000) to W.X.Z.
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  • Sperm are specialized cells that require adenosine triphosphate (ATP) to support their function. Maintaining sperm energy homeostasis in vitro is vitally important to improve the efficacy of boar sperm preservation. Metformin can activate 5′-AMP-activated protein kinase (AMPK) to improve metabolic flexibility and maintain energy homeostasis. Thus, the aim of the present study was to investigate whether metformin can improve boar sperm quality through AMPK mediation of energy metabolism. Sperm motility parameters, membrane integrity, acrosome integrity, mitochondrial membrane potential (ΔΨm), ATP content, glucose uptake, and lactate efflux were analyzed. Localization and expression levels of AMPK and phospho-Thr172-AMPK (p-AMPK) were also detected by immunofluorescence and western blotting. We found that metformin treatment significantly increased sperm motility parameters, ΔΨm, and ATP content during storage at 17 °C. Moreover, results showed that AMPK was localized at the acrosomal region, connecting piece, and midpiece of sperm and p-AMPK was distributed at the post-acrosomal region, connecting piece, and midpiece. When sperm were incubated with metformin for 4 h at 37 °C, sperm motility parameters, ΔΨm, ATP content, p-AMPK, glucose uptake, and lactate efflux all significantly increased, whereas the addition of Compound C treatment, an inhibitor of AMPK, counteracted these positive effects. Together, our results suggest that metformin promotes AMPK activation, which contributes to the maintenance of energy hemostasis and mitochondrial activity, thereby maintaining boar sperm functionality and improving the efficacy of semen preservation.
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  • [1]
    Berstein LM. 2012. Metformin in obesity, cancer and aging: addressing controversies. Aging (Albany NY), 4(5): 320−329.
    [2]
    Bertoldo MJ, Guibert E, Tartarin P, Guillory V, Froment P. 2014. Effect of metformin on the fertilizing ability of mouse spermatozoa. Cryobiology, 68(2): 262−268. doi: 10.1016/j.cryobiol.2014.02.006
    [3]
    Bielas W, Nizanski W, Partyka A, Rzasa A, Mordak R. 2017. Effect of long-term storage in Safe Cell+ extender on boar sperm DNA integrity and other key sperm parameters. Acta Veterinaria Scandinavica, 59(1): 58. doi: 10.1186/s13028-017-0325-9
    [4]
    Brand KA, Hermfisse U. 1997. Aerobic glycolysis by proliferating cells: a protective strategy against reactive oxygen species. FASEB Journal, 11(5): 388−395. doi: 10.1096/fasebj.11.5.9141507
    [5]
    Brunmair B, Staniek K, Gras F, Scharf N, Althaym A, Clara R, et al. 2004. Thiazolidinediones, like metformin, inhibit respiratory complex I: a common mechanism contributing to their antidiabetic actions?. Diabetes, 53(4): 1052−1059. doi: 10.2337/diabetes.53.4.1052
    [6]
    Calle-Guisado V, de Llera AH, Martin-Hidalgo D, Mijares J, Gil MC, Alvarez IS, et al. 2017. AMP-activated kinase in human spermatozoa: identification, intracellular localization, and key function in the regulation of sperm motility. Asian Journal of Andrology, 19(6): 707−714. doi: 10.4103/1008-682X.185848
    [7]
    Chandel NS, Avizonis D, Reczek CR, Weinberg SE, Menz S, Neuhaus R, et al. 2016. Are metformin doses used in murine cancer models clinically relevant?. Cell Metabolism, 23(4): 569−570. doi: 10.1016/j.cmet.2016.03.010
    [8]
    Detaille D, Guigas B, Chauvin C, Batandier C, Fontaine E, Wiernsperger N, et al. 2005. Metformin prevents high-glucose-induced endothelial cell death through a mitochondrial permeability transition-dependent process. Diabetes, 54(7): 2179−2187. doi: 10.2337/diabetes.54.7.2179
    [9]
    El-Mir MY, Nogueira V, Fontaine E, Avéret N, Rigoulet M, Leverve X. 2000. Dimethylbiguanide inhibits cell respiration via an indirect effect targeted on the respiratory chain complex I. Journal of Biological Chemistry, 275(1): 223−228. doi: 10.1074/jbc.275.1.223
    [10]
    Foretz M, Viollet B. 2011. Regulation of hepatic metabolism by AMPK. Journal of Hepatology, 54(4): 827−829. doi: 10.1016/j.jhep.2010.09.014
    [11]
    Fu J, Li Y, Wang L, Zhen L, Yang Q, Li P, et al. 2017. Bovine serum albumin and skim-milk improve boar sperm motility by enhancing energy metabolism and protein modifications during liquid storage at 17 °C. Theriogenology, 102: 87. doi: 10.1016/j.theriogenology.2017.07.020
    [12]
    Gravel SP, Hulea L, Toban N, Birman E, Blouin MJ, Zakikhani M, et al. 2014. Serine deprivation enhances antineoplastic activity of biguanides. Cancer Research, 74(24): 7521−7533. doi: 10.1158/0008-5472.CAN-14-2643-T
    [13]
    Han JJ, Zhang L, Guo H, Wysham WZ, Roque DR, Willson AK, et al. 2015. Glucose promotes cell proliferation, glucose uptake and invasion in endometrial cancer cells via AMPK/mTOR/S6 and MAPK signaling. Gynecologic Oncology, 138(3): 668−675. doi: 10.1016/j.ygyno.2015.06.036
    [14]
    Hardie DG. 2011. AMP-activated protein kinase-an energy sensor that regulates all aspects of cell function. Genes and Development, 25(18): 1895−1908. doi: 10.1101/gad.17420111
    [15]
    Hardie DG, Ross FA, Hawley SA. 2012. AMPK: a nutrient and energy sensor that maintains energy homeostasis. Nature Reviews Molecular Cell Biology, 13(4): 251−262. doi: 10.1038/nrm3311
    [16]
    Hirohashi N, Tamura-Nakano M, Nakaya F, Iida T, Iwata Y. 2016. Sneaker male squid produce long-lived spermatozoa by modulating their energy metabolism. Journal of Biological Chemistry, 291(37): 19324−19334. doi: 10.1074/jbc.M116.737494
    [17]
    Hurtado de Llera A, Martin-Hidalgo D, Garcia-Marin LJ, Bragado MJ. 2018. Metformin blocks mitochondrial membrane potential and inhibits sperm motility in fresh and refrigerated boar spermatozoa. Reproduction in Domestic Animals, 53(3): 733−741. doi: 10.1111/rda.13164
    [18]
    Hurtado de Llera A, Martin-Hidalgo D, Gil MC, Garcia-Marin LJ, Bragado MJ. 2012. AMP-activated kinase AMPK is expressed in boar spermatozoa and regulates motility. PloS One, 7(6): e38840. doi: 10.1371/journal.pone.0038840
    [19]
    Hurtado de Llera A, Martin-Hidalgo D, Rodriguez-Gil JE, Gil MC, Garcia-Marin LJ, Bragado MJ. 2013. AMP-activated kinase, AMPK, is involved in the maintenance of plasma membrane organization in boar spermatozoa. Biochimica et Biophysica Acta (BBA) - Biomembranes, 1828(9): 2143−2151. doi: 10.1016/j.bbamem.2013.05.026
    [20]
    Kondoh H, Lleonart ME, Gil J, Wang J, Degan P, Peters G, et al. 2005. Glycolytic enzymes can modulate cellular life span. Cancer Research, 65(1): 177−185.
    [21]
    Lin SC, Hardie DG. 2018. AMPK: Sensing glucose as well as cellular energy status. Cell Metabolism, 27(2): 299−313. doi: 10.1016/j.cmet.2017.10.009
    [22]
    Lv YH, Zhang PF, Guo JY, Zhu ZD, Li XL, Xu D, et al. 2018. Melatonin protects mouse spermatogonial stem cells against hexavalent chromium-induced apoptosis and epigenetic histone modification. Toxicology and Applied Pharmacology, 340: 30−38. doi: 10.1016/j.taap.2017.12.017
    [23]
    Martin-Hidalgo D, Hurtado de Llera A, Calle-Guisado V, Gonzalez-Fernandez L, Garcia-Marin L, Bragado MJ. 2018. AMPK function in mammalian spermatozoa. International Journal of Molecular Sciences, 19(11): 3293. doi: 10.3390/ijms19113293
    [24]
    Martin-Montalvo A, Mercken EM, Mitchell SJ, Palacios HH, Mote PL, Scheibye-Knudsen M, et al. 2013. Metformin improves healthspan and lifespan in mice. Nature Communications, 4(1): 2192. doi: 10.1038/ncomms3192
    [25]
    Nguyen TMD. 2017. Impact of 5′-AMP-activated protein kinase on male gonad and spermatozoa functions. Frontiers in Cell and Developmental Biology, 5: 25.
    [26]
    Nguyen TMD. 2019. Role of AMPK in mammals reproduction: Specific controls and whole-body energy sensing. Comptes Rendus Biologies, 342(1-2): 1−6. doi: 10.1016/j.crvi.2018.10.003
    [27]
    Nguyen TMD, Alves S, Grasseau I, Metayer-Coustard S, Praud C, Froment P, et al. 2014. Central role of 5′-AMP-activated protein kinase in chicken sperm functions. Biology of Reproduction, 91(5): 121.
    [28]
    Nguyen TMD, Seigneurin F, Froment P, Combarnous Y, Blesbois E. 2015. The 5′-AMP-activated protein kinase (AMPK) is involved in the augmentation of antioxidant defenses in cryopreserved chicken sperm. PloS One, 10(7): e0134420. doi: 10.1371/journal.pone.0134420
    [29]
    Polianskyte-Prause Z, Tolvanen TA, Lindfors S, Dumont V, Van M, Wang H, et al. 2019. Metformin increases glucose uptake and acts renoprotectively by reducing SHIP2 activity. FASEB Journal, 33(2): 2858−2869. doi: 10.1096/fj.201800529RR
    [30]
    Rodríguez-Gil JE. 2006. Mammalian sperm energy resources management and survival during conservation in refrigeration. Reproduction in Domestic Animals, 41(S2): 11−20. doi: 10.1111/j.1439-0531.2006.00765.x
    [31]
    Rodríguez-Gil JE, Bonet S. 2016. Current knowledge on boar sperm metabolism: Comparison with other mammalian species. Theriogenology, 85(1): 4−11. doi: 10.1016/j.theriogenology.2015.05.005
    [32]
    Shabani Nashtaei M, Amidi F, Sedighi Gilani MA, Aleyasin A, Bakhshalizadeh S, Naji M, et al. 2017. Protective features of resveratrol on human spermatozoa cryopreservation may be mediated through 5' AMP-activated protein kinase activation. Andrology, 5(2): 313−326. doi: 10.1111/andr.12306
    [33]
    Swegen A, Lambourne SR, Aitken RJ, Gibb Z. 2016. Rosiglitazone improves stallion sperm motility, ATP content, and mitochondrial function. Biology of Reproduction, 95(5): 107. doi: 10.1095/biolreprod.116.142687
    [34]
    Tartarin P, Guibert E, Touré A, Ouiste C, Leclerc J, Sanz N, et al. 2012. Inactivation of AMPKα1 induces asthenozoospermia and alters spermatozoa morphology. Endocrinology, 153(7): 3468−3481. doi: 10.1210/en.2011-1911
    [35]
    Tsuda Y, Iwasawa K, Yokoyama M, Yamaguchi M. 2017. Trypsin-treated β-lactoglobulin improves glucose tolerance in C57BL/6 Mice by enhancing AMPK activation and glucose uptake in hepatocytes. Biological and Pharmaceutical Bulletin, 40(11): 1917−1922. doi: 10.1248/bpb.b17-00437
    [36]
    Ushiyama A, Priyadarshana C, Setiawan R, Miyazaki H, Ishikawa N, Tajima A, et al. 2019. Membrane raft-mediated regulation of glucose signaling pathway leading to acrosome reaction in chicken sperm. Biology of Reproduction, 100(6): 1482−1491. doi: 10.1093/biolre/ioz015
    [37]
    Vazirian M, Nabavi SM, Jafari S, Manayi A. 2018. Natural activators of adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) and their pharmacological activities. Food and Chemical Toxicology, 122: 69−79. doi: 10.1016/j.fct.2018.09.079
    [38]
    Vial G, Detaille D, Guigas B. 2019. Role of mitochondria in the mechanism(s) of action of metformin. Frontiers in Endocrinology, 10: 294. doi: 10.3389/fendo.2019.00294
    [39]
    Vlavcheski F, Naimi M, Murphy B, Hudlicky T, Tsiani E. 2017. Rosmarinic acid, a rosemary extract polyphenol, increases skeletal muscle cell glucose uptake and activates AMPK. Molecules, 22(10): 1669. doi: 10.3390/molecules22101669
    [40]
    Waberski D, Riesenbeck A, Schulze M, Weitze KF, Johnson L. 2019. Application of preserved boar semen for artificial insemination: Past, present and future challenges. Theriogenology, 137: 2−7. doi: 10.1016/j.theriogenology.2019.05.030
    [41]
    Xu L, Kong L, Wang J, Ash JD. 2018. Stimulation of AMPK prevents degeneration of photoreceptors and the retinal pigment epithelium. Proceedings of the National Academy of Sciences of the United States of America, 115(41): 10475−10480. doi: 10.1073/pnas.1802724115
    [42]
    Zhu ZD, Kawai T, Umehara T, Hoque SAM, Zeng WX, Shimada M. 2019a. Negative effects of ROS generated during linear sperm motility on gene expression and ATP generation in boar sperm mitochondria. Free Radical Biology and Medicine, 141: 159−171. doi: 10.1016/j.freeradbiomed.2019.06.018
    [43]
    Zhu ZD, Li RN, Fan XT, Lv YH, Zheng Y, Hoque SAM, et al. 2019b. Resveratrol improves boar sperm quality via 5'AMP-activated protein kinase activation during cryopreservation. Oxidative Medicine and Cellular Longevity, 2019: 5921503.
    [44]
    Zhu ZD, Li RN, Ma GZ, Bai WJ, Fan XT, Lv YH, et al. 2018. 5′-AMP-activated protein kinase regulates goat sperm functions via energy metabolism in vitro. Cellular Physiology and Biochemistry, 47(6): 2420−2431. doi: 10.1159/000491616
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