Kisspeptin-mediated regulation of testicular activity of rats under the effect of gold nanoparticles

  • V. Y. Kalynovskyi Taras Shevchenko National University of Kyiv
  • A. S. Pustovalov Taras Shevchenko National University of Kyiv
  • G. Y. Grodzyuk Nanomedtech-LLC
  • N. S. Andriushyna Nanomedtech-LLC
  • M. E. Dzerzhynsky Taras Shevchenko National University of Kyiv
Keywords: nanogold, Sertoli cells, Leydig cells, kisspeptin-10, peptide-234


There are a variety of biomedical applications of nanoparticles. They can be used as drug carriers, anti-tumor agents, biosensors and modulators of immune response. But full-scale real clinical application of nanomaterials requires a great deal of information on their safety and biotoxicity. Even traditionally harmless materials, like gold, can obtain toxic features when scaled to the nanosize. In vitro studies showed that nanoparticles can be geno- and cytotoxic, but their effects on the body as a whole remain largely a mystery. To shed some light on this, our study focused on the reproductive toxicity of nanomaterials. We synthesized 10–15 nm gold nanoparticles through the reduction of sodium tetrachloroaurate (III) in an alkaline medium with the addition of sodium polyphosphate as a stabilizing agent. Next, these particles were administered intraperitoneally to young and old rats for 10 days. To test functional capabilities of the testes, we injected kisspeptin-10 or its antagonist peptide-234 intracerebroventricularly. These substances are known to stimulate or inhibit the central component of the hypothalamic-pituitary-gonadal axis respectively. After the routine histological procedures, we measured the diameter of seminiferous tubules and the nuclear cross-sectional area of Sertoli cells as markers of testicular spermatogenic activity and a cross-sectional area of the Leydig cells’ nuclei as a marker of testicular steroidogenesis. We found that injections of nanogold caused no significant changes in the young animals. At the same time, morphometric parameters of adult animals were significantly lower compared to control, although we observed no pathological changes in the tissue. Combined administration of gold nanoparticles and kisspeptin showed that the stimulatory effect of the latter was not observed at all. This is a specific feature of toxicants called “endocrine disruptors”. Moreover, we found morphological signs of testicular degeneration, which are characteristic of the low-testosterone state. Simultaneous injections of gold and peptide-234 resulted in the highest degree of testicular functional downregulation, regardless of age. Taken as a whole, our data indicates that gold nanoparticles disrupt the regulatory network of the hypothalamic-pituitary-gonadal axis, possibly due to direct action on the interstitial cells and spermatogenic epithelium. 


Asare, N., Instanes, C., Sandberg, W., Refsnes, M., Schwarze, P., Kruszewski, M., Brunborg, G., 2012. Cytotoxic and genotoxic effects of silver nanoparticles in testicular cells. Toxicology 291, 65–72. >>
Balasubramanian, S., Jittiwat, J., Manikandan, J., Ong, C.-N., Yu, L., Ong, W.-Y., 2010. Biodistribution of gold nanoparticles and gene expression changes in the liver and spleen after intravenous administration in rats. Biomaterials 31(8), 2034–2042. >>
Bogdanoviс, G., Djordjeviс, A., 2016. Carbon nanomaterials: Biologically active fullerene derivatives. Srp. Arh. Celok. Lek. 144, 222–231. >>
Buerki-Thurnherr, T., Mandach, U., Wick, P., 2012. Knocking at the door of the unborn child: Engineered nanoparticles at the human placental barrier. Swiss Medical Weekly 142, w13559. >>
Cabuzu, D., Cirja, A., Puiu, R., Grumezescu, A., 2015. Biomedical applications of gold nanoparticles. Curr. Top. Med. Chem. 15(16), 1605–1613. >>
Creasy, D., Bube, A., Rijk, E., Kandori, H., Kuwahara, M., Masson, R., Nolte, T., Reams, R., Regan, K., Rehm, S., Rogerson, P., Whitney, K., 2012. Proliferative and nonproliferative lesions of the rat and mouse male reproductive system. Toxicol. Pathol. 40(6), 40S-121S. >>
Dreaden, E., Alkilany, A., Huang, X., Murphy, C., El-Sayed, M., 2011. The golden age: Gold nanoparticles for biomedicine. Chem. Soc. Rev. 41(7), 2740–2779. >>
Gao, G., Ze, Y., Zhao, X., Sang, X., Zheng, L., Ze, X., Gui, S., Sheng, L., Sun, Q., Hong, J., Yu, X., Wang, L., Hong, F., Zhang, X., 2013. Titanium dioxide nanoparticle-induced testicular damage, spermatogenesis suppression, and gene expression alterations in male mice. J. Hazard. Mater. 258–259, 133–143. >>
Han, J., Jeong, J.-K., Gurunathan, S., Choi, Y.-J., Das, J., Kwon, D.-N., Cho, S.-G., Park, C., Seo, H., Park, J., Kim, J., 2016. Male- and female-derived somatic and germ cellspecific toxicity of silver nanoparticles in mouse. Nanotoxicology 10(3), 361–373. >>
Helena, C., Toporikova, N., Kalil, B., Stathopoulos, A., Pogrebna, V., Carolino, R., Anselmo-Franci, J., Bertram, R., 2015. KNDy neurons modulate the magnitude of the steroid-induced luteinizing hormone surges in ovariectomized rats. Endocrinology 156(11), 4200–4213. >>
Kalynovskyi, V., Pustovalov, A., Grodzyuk, G., Andriushyna, N., Dzerzhynsky, M., 2016. Vplyv nanochastynok ta ioniv zolota na morfo-funkcionalnyi stan sim’janykiv statevonezrilyh shhuriv [Testicular morpho-functional state of immature rats under the effect of gold nanoparticles and ions]. Visn. Kyiv Nat. Univ. im. Tar. Shev. Ser. Biol. 71(1), 23–26 (in Ukrainian).
Kim, J.-E., Shin, J.-Y., Cho, M.-H., 2012. Magnetic nanoparticles: An update of application for drug delivery and possible toxic effects. Arch. Toxicol. 86(5), 685–700. >>
Li, W., Wang, F., Liu, Z., Wang, Y., Wang, J., Sun, F., 2013. Gold nanoparticles elevate plasma testosterone levels in male mice without affecting fertility. Small 9, 1708–1714. >>
Lu, X., Liu, Y., Kong, X., Lobie, P., Chen, C., Zhu, T., 2013. Nanotoxicity: A growing need for study in the endocrine system. Small 9, 1654–1671. >>
Lucas, B., Fields, C., Hofmann, M.-C., 2009. Signaling pathways in spermatogonial stem cells and their disruption by toxicants. Birth Defects Research. Part C, Embryo Today: Reviews 87(1), 35–42. >>
Matvienko, M.G., Pustovalov, A.S., Dzerzhinsky, M.E., 2013. Variety of functions and effects of kisspeptin. Biopolym. Cell 29(1), 11–20. >>
Melnik, E., Buzulukov, Y., Demin, V., Demin, V., Gmoshinski, I., Tyshko, N., Tutelyan, V., 2013. Transfer of silver nano-particles through the placenta and breast milk during in vivo experiments on rats. Acta Naturae 5(3), 107–115.
Paxinos, G., Watson, C., 2007. The rat brain in stereotaxic coordinates 6th edition. Academic Press, London.
Pineda, R., Garcia-Galiano, D., Roseweir, A., Romero, M., Sanchez-Garrido, M.A., Ruiz-Pino, F., Morgan, K., Pinilla, L., Millar, R., Tena-Sempere, M., 2010. Critical roles of kisspeptins in female puberty and preovulatory gonadotropin surges as revealed by a novel antagonist. Endocrinology 151(2), 722–730. >>
Pizent, A., Tariba, B., Živković, T., 2012. Reproductive toxicity of metals in men. Archives of Industrial Hygiene and Toxicology 63(Suppl. 1), 35–46. >>
Reyes-Esparza, J., Martínez-Mena, A., Gutiérrez-Sancha, I., Rodríguez-Fragoso, P., de la Cruz, G.G., Mondragón, R., Rodríguez-Fragoso, L., 2015. Synthesis, characterization and biocompatibility of cadmium sulfide nanoparticles capped with dextrin for in vivo and in vitro imaging application. J. Nanobiotechnology 13, 83. >>
Shao, J., Griffin, R., Galanzha, E., Kim, J.-W., Koonce, N., Webber, J., Mustafa, T., Birls, A., Nedosekin, D., Zharov, V., 2013. Photothermal nanodrugs: Potential of TNF-gold nano-spheres for cancer theranostics. Sci. Rep. 3, 1293. >>
Talebi, A., Khorsandi, L., Moridian, M., 2013. The effect of zinc oxide nanoparticles on mouse spermatogenesis. J. Assist. Reprod. Genet. 30(9), 1203–1209. >>
Thakur, M., Gupta, H., Singh, D., Mohanty, I., Maheswari, U., Vanage, G., Joshi, D., 2014. Histopathological and ultra structural effects of nanoparticles on rat testis following 90 days (chronic study) of repeated oral administration. J. Nano-biotechnology 12, 42. >>
Wang, L., Chen, C., 2016. Pathophysiologic mechanisms of bio-medical nanomaterials. Toxicol. Appl. Pharm. 299, 30–40. >>
Zakhidov, S., Pavliuchenkova, S., Marshak, T., Rudoi, V., Dement’eva, O., Zelenina, I., Skuridin, S., Makarov, A., Khokhlov, A., Evdokimov, I., 2012. Effect of gold nanoparticles on mouse spermatogenesis. Izv. Akad. Nauk Ser. Biol. 39(3), 279–287. >>
Zande, M., Vandebriel, R., Doren, E., Kramer, E., Rivera, Z., Serrano-Rojero, C., Gremmer, E., Mast, J., Peters, R., Hollman, P., Hendriksen, P., Marvin, H., Peijnenburg, A., Bouwmeester, H., 2012. Distribution, elimination, and toxicity of silver nanoparticles and silver ions in rats after 28-day oral exposure. ACS Nano 6(8), 7427–7442. >>