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Effects of Boron on Cytotoxicity, Apoptosis, and Cell Cycle of Cultured Rat Sertoli Cells In vitro

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Abstract

The present study aimed to investigate the effects of the administration of boron on viability, apoptosis, and cell cycle of primary rat Sertoli cells (SCs) in vitro. SCs were aseptically isolated from 18–22-day-old male Sprague-Dawley (SD) rats. SCs were identified with immunofluorescence using anti-vimentin antibody. Further, to investigate the effects of boron on Sertoli cells, SCs of the boron treatment group were exposed to different concentrations (0.25, 0.5, 1, 5, 10, 40, and 80 mmol/L) of boric acid. Using MTT and Cell Counting Kit-8 assays, the impact of boron on SCs viability was analyzed. Cell apoptosis and cycle of SCs were analyzed using flow cytometry. A concentration of 0.5 mmol/L boric acid resulted in the highest viability and lowest necrosis and apoptosis. Above this concentration (even 1.0 mmol/L) showed lower viability and higher levels of necrosis and apoptosis. Administration of < 0.5 mmol/L boron significantly promoted the viability of Sertoli cells (P < 0.01); however, the exposure to high dose (> 10 mmol/L) of boron exhibited significant adverse effects on Sertoli cells (P < 0.01) and even toxic effects, inhibiting cell viability compared to the control group. Flow cytometry analysis showed that treatment with 0.5 mmol/L of boron significantly inhibited the apoptosis of Sertoli cells and the proportion of cells in S and G2/M phases was markedly increased; however, a higher concentration of 40 and 80 mmol/L of boron promoted Sertoli cell apoptosis and cells were completely arrested at G0/G1 phase. Boron at doses below 0.5 mmol/L could significantly improve the viable capacity of testicular Sertoli cells in vitro and inhibit their apoptosis. However, high dose of boron (at a concentration higher than 5.0 mmol/L) exhibited noticeable toxic effects, inhibiting cell viability, accelerating apoptosis of Sertoli cells, and arresting cell cycle at G0/G1 phase.

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Abbreviations

SCs:

Sertoli cells

SD:

Sprague-Dawley

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

Fetal bovine serum

PBS:

Phosphate-buffered saline

PFA:

Paraformaldehyde

DAB:

3,3′-Diaminobenzidine

PI:

Propidium iodide

References

  1. Oganov AR, Ma Y, Lyakhov AO, Valle M, Gatti C (2010) Evolutionary crystal structure prediction as a method for the discovery of minerals and materials. Rev Mineral Geochem 71:271–298. https://doi.org/10.2138/rmg.2010.71.13

    Article  CAS  Google Scholar 

  2. Samman S, Naghii MR, Lyons Wall PM, Verus AP (1998) The nutritional and metabolic effects of boron in humans and animals. Biol Trace Elem Res 66(1-3):227–235. https://doi.org/10.1007/BF02783140

    Article  CAS  PubMed  Google Scholar 

  3. Hu Q, Li S, Qiao E, Gu Y (2014) Effects of boron on structure and antioxidative activities of spleen in rats. Biol Trace Elem Res 158:73–80. https://doi.org/10.1007/s12011-014-9899-5

    Article  CAS  PubMed  Google Scholar 

  4. Jin E, Gu Y, Wang J, Jin G, Li S (2014) Effect of supplementation of drinking water with different levels of boron on performance and immune organ parameters of broilers. Ital J Anim Sci 13:215–214. https://doi.org/10.4081/ijas.2014.3152

    Article  Google Scholar 

  5. Jin E, Li S, Ren M, Hu Q, Gu Y, Li K (2017) Boron affects immune function through modulation of splenic T lymphocyte subsets, cytokine secretion, and lymphocyte proliferation and apoptosis in rats. Biol Trace Elem Res 17:8261–8275. https://doi.org/10.1007/s12011-017-0932-3

    Article  CAS  Google Scholar 

  6. Xiao K, Yang K, Wang J, Sun P, Huang H, Khaliq H, Peng K (2018) Transcriptional study revealed that boron supplementation may alter the immune-related genes through MAPK signaling in ostrich chick thymus. Biol Trace Elem Res 189(1):209–223. https://doi.org/10.1007/s12011-018-1441-8

    Article  CAS  PubMed  Google Scholar 

  7. Hunt C (2008) Dietary boron: possible roles in human and animal physiology. Biomed Res Trace Elem 19:243–253. https://doi.org/10.11299/brte.19.243

    Article  CAS  Google Scholar 

  8. Duydu Y, Başaran N, Üstündağ A, Aydın S, Ündeğer Ü, Ataman OY, Bolt HM (2011) Reproductive toxicity parameters and biological monitoring in occupationally and environmentally boron-exposed persons in Bandirma, Turkey. Arch Toxicol 85(6):589–600. https://doi.org/10.1007/s00204-011-0692-3

    Article  CAS  PubMed  Google Scholar 

  9. Robbins WA, Xun L, Jia J, Kennedy N, Elashoff DA, Ping L (2010) Chronic boron exposure and human semen parameters. Reprod Toxicol 29(2):184–190. https://doi.org/10.1016/j.reprotox.2009.11.003

    Article  CAS  PubMed  Google Scholar 

  10. Duydu Y, Ustundag A, Yalcin CO, Bolt HM (2017) Investigation of boron mediated reproductive and developmental effects in highly boron exposed population. Toxicol Lett 280:s94. https://doi.org/10.1016/j.toxlet.2017.07.259

    Article  Google Scholar 

  11. Meroni SB, Galardo MN, Rindone G, Gorga A, Riera MF, Cigorraga SB (2019) Molecular mechanisms and signaling pathways involved in sertoli cell proliferation frontiers in endocrinology, 10. https://doi.org/10.3389/fendo.2019.00224

  12. Johnson L, Thompson DL (2007) Role of Sertoli cell number and function on regulation of spermatogenesis. Anim Reprod Sci 2008(105):23–51. https://doi.org/10.1016/j.anireprosci.2007.11.029

    Article  CAS  Google Scholar 

  13. Krishnan BB, Selvaraju S, Gowda NKS, Subramanya KB, Pal D, Archana SS, Bhatta R (2019) Dietary boron supplementation enhances sperm quality and immunity through influencing the associated biochemical parameters and modulating the genes expression at testicular tissue. J Trace Elem Med Biol. https://doi.org/10.1016/j.jtemb.2019.05.004

  14. Ibrahim TB, Abdel-Wahab A, El-Anwar AH, Ibrahim SS (2019) Dietary boron supplementation and its impact on testicular function, thyroid activity and serum calcium in rams. Small Rumin Res. https://doi.org/10.1016/j.smallrumres.2019.03.008

  15. Jin E, Hu Q, Ren M, Chen X, Li S, Xu W (2016) Effects of boron supplementation on hepatic testis and epididymis in male obese rats. Acta Nutrimenta Sinica 38:466–472. https://doi.org/10.13325/j.cnki.acta.nutr.sin.2016.05.015

    Article  Google Scholar 

  16. Sriram A, Huynh HDN, Shadarevian J, Djaksigulova D, Vogl AW (2017) Primary Sertoli cell cultures from 30-day old rats form cords with a morphologically differentiated phenotype. FASEB J 31(1-supplement):740.4

    Google Scholar 

  17. Mai Q, Xie J, Huang R (2007) Research status and application prospect of testicular Sertoli cells. Chinese J Tissue Eng Res 11(43):8761–8764. https://doi.org/10.3321/j.issn:1673-8225.2007.43.017

  18. Zhang J (1996) Practical methods and techniques in cell biology [M]. Beijing: Joint press of Beijing medical university and China union medical college 1996:8–10

    Google Scholar 

  19. Dufour JM, Rajotte RV, Seeberger K, Kin T, Korbutt GS (2003) Long-term survival of neonatal porcine Sertoli cells in non-immunosuppressed rats. Xenotransplantation 10(6):577–586. https://doi.org/10.1034/j.1399-3089.2003.00059.x

    Article  PubMed  Google Scholar 

  20. Duan X, Song C, Xu P et al (2004) Separation and identification of rat FasL-expressing Sertoli cells. Basic Med Sci Clin 24:88–91

  21. Fort DJ (2002) Boron deficiency disables Xenopus laevis oocyte maturation events. Biol Trace Elem Res 85:157–169. https://doi.org/10.1385/BTER:85:2:157

    Article  CAS  PubMed  Google Scholar 

  22. Armstrong TA, Flowers WL, Spears JW (2002) Long-term effects of boron supplementation on reproductive characteristics and bone mechanical properties in gilts. J Anim Sci 80(1):154–161. https://doi.org/10.2527/2002.801154x

    Article  CAS  PubMed  Google Scholar 

  23. Li S (2008) Effect and mechanism of boron on nutrition and toxicity in rats [D]. Huazhong Agricultural University

  24. Bustos-Obregon E, Carvallo M, Hartley-Belmar R (2007) Histopathological and histometrical assessment of boron exposure effects on mouse spermatogenesis. Int J Morphol 25(4):919–925. https://doi.org/10.4067/s0717-95022007000400039

    Article  Google Scholar 

  25. Lucas TFG, Pimenta MT, Pisolato R, Lazari MFM, Porto CS (2011) 17β-estradiol signaling and regulation of Sertoli cell function. Spermatogenesis 1(4):318–324. https://doi.org/10.4161/spmg.1.4.18903

    Article  PubMed  PubMed Central  Google Scholar 

  26. Walker WH (2003) Molecular mechanisms controlling Sertoli cell proliferation and differentiation. Endocrinology 144(9):3719–3721. https://doi.org/10.1210/en.2003-0765

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the National Natural Science Foundation of China (31672502, 31502137, 31501968, and 31402154), the Major Science and Technology Projects in Anhui Province (17030701004), the Major Projects of the Education Department of Anhui Province (KJ2017ZD43), Anhui Provincial Natural Science Foundation (Grant No. 1608085QC72), the Foundation of Anhui Science and Technology University (ZRC2013354, ZRC2014453), the first-class cultivation subject of veterinary science in Anhui Science and Technology University, The Key Program in the Youth Elite Support Plan in Universities of Anhui Province (gxyqZD2019061).

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  1. Liangyue Lu and Qian Zhang contributed equally to this work.

Authors and Affiliations

  1. College of Animal Science, Anhui Science and Technology University, No.9 Donghua Road, Fengyang County, 233100, Anhui Province, People’s Republic of China

    Liangyue Lu, Qian Zhang, Man Ren, Erhui Jin, Qianqian Hu, Chunfang Zhao & Shenghe Li

  2. Key Laboratory of Quality & Safety Control for Pork, Ministry of Agriculture and Rural, Fengyang County, Anhui Province, People’s Republic of China

    Man Ren, Erhui Jin, Qianqian Hu, Chunfang Zhao & Shenghe Li

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  1. Liangyue Lu
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Lu, L., Zhang, Q., Ren, M. et al. Effects of Boron on Cytotoxicity, Apoptosis, and Cell Cycle of Cultured Rat Sertoli Cells In vitro. Biol Trace Elem Res 196, 223–230 (2020). https://doi.org/10.1007/s12011-019-01911-3

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