Skip to main content

Advertisement

SpringerLink
Log in

Effects of HIF-1α on Spermatogenesis of Varicocele Rats by Regulating VEGF/PI3K/Akt Signaling Pathway

  • Male Reproduction: Original Article
  • Published:
Reproductive Sciences Aims and scope Submit manuscript

Abstract

Hypoxia-inducible factor-1α (HIF-1α) participates in the regulation of spermatogenic function in rats with varicocele (VC), and the PI3K/Akt pathway plays an important role in it. In the present research, we applied the CRISPR/Cas9 gene editing technique to silence the HIF-1α gene of VC rat testis, to explore the effect of HIF-1α on apoptosis of spermatogenic cells in VC rats through the PI3K/Akt pathway. Sprague Dawley rats were randomly assigned to four groups, including the normal rat group (group N), VC model group (group V), VC + HIF-1α-lentivirus group (group H), and VC + luciferase-lentivirus group (group L). Apoptosis of spermatogenic cells in rat testis was tested by TUNEL Kit. The morphologic changes of seminiferous tubules were viewed by a light microscope. Expressions of VEGF, Akt, p-Akt, p70S6K, and p-p70S6K were detected by means of Western blot, immunofluorescence, or immunohistochemistry methods. One-way ANOVA was applied to analyze the diverseness between groups. Compared with group N, the distribution of germ cells was disordered, apoptosis of spermatogenic cells increased significantly, and the expression of VEGF, p-Akt, and p-p70S6K was also increased in group V. Compared with group V, the damage of seminiferous epithelium in group H was improved, and the arrangement of the seminiferous epithelium was almost orderly. Apoptosis of spermatogenic cells decreased significantly, and the expression of VEGF, p-Akt, and p-p70S6K protein was decreased (P < 0.05). There was no significant difference between group N and group H (P > 0.05).

In conclusion, HIF-1α is regulated by hypoxia in rats with varicocele to regulate its downstream gene VEGF which regulates spermatogenesis, and the PI3K/Akt signaling pathway plays a regulatory role in this process.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

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

Similar content being viewed by others

Data Availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

References

  1. Pastuszak A, Wang R. Varicocele and testicular function. Asian J Androl. 2015;17(4):659. https://doi.org/10.4103/1008-682x.153539.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Guo WB, Yang C, Bian J, Xia H, Yang JK, Zhou QZ, et al. With a new clip technique surgically inducing varicocele in Sprague-Dawley rats. BMC Urol. 2018;18(1):58. https://doi.org/10.1186/s12894-018-0350-7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Kaminsky A, Sperling H. Variocele in adolescents. Urologe A. 2014;53(2):213–7. https://doi.org/10.1007/s00120-013-3384-1.

    Article  CAS  PubMed  Google Scholar 

  4. Kumanov P, Robeva RN, Tomova A. Adolescent varicocele: who is at risk? Pediatrics. 2008;121(1):e53–7. https://doi.org/10.1542/peds.2007-0340.

    Article  PubMed  Google Scholar 

  5. Agarwal A, Hamada A, Esteves SC. Insight into oxidative stress in varicocele-associated male infertility: part 1. Nat Rev Urol. 2012;9(12):678–90. https://doi.org/10.1038/nrurol.2012.197.

    Article  CAS  PubMed  Google Scholar 

  6. Sheehan MM, Ramasamy R, Lamb DJ. Molecular mechanisms involved in varicocele-associated infertility. J Assist Reprod Genet. 2014;31(5):521–6. https://doi.org/10.1007/s10815-014-0200-9.

    Article  PubMed  PubMed Central  Google Scholar 

  7. Shiraishi K, Matsuyama H, Takihara H. Pathophysiology of varicocele in male infertility in the era of assisted reproductive technology. Int J Urol. 2012;19(6):538–50. https://doi.org/10.1111/j.1442-2042.2012.02982.x.

    Article  CAS  PubMed  Google Scholar 

  8. Wang H, Sun Y, Wang L, Xu C, Yang Q, Liu B, et al. Hypoxia-induced apoptosis in the bilateral testes of rats with left-sided varicocele: a new way to think about the varicocele. J Androl. 2010;31(3):299–305. https://doi.org/10.2164/jandrol.108.007153.

    Article  PubMed  Google Scholar 

  9. Wang H, Lv Y, Hu K, Feng T, Jin Y, Wang Y, et al. Seminal plasma leptin and spermatozoon apoptosis in patients with varicocele and leucocytospermia. Andrologia. 2015;47(6):655–61. https://doi.org/10.1111/and.12313.

    Article  CAS  PubMed  Google Scholar 

  10. Ghandehari-Alavijeh R, Tavalaee M, Zohrabi D, Foroozan-Broojeni S, Abbasi H, Nasr-Esfahani MH. Hypoxia pathway has more impact than inflammation pathway on etiology of infertile men with varicocele. Andrologia. 2019;51(2):e13189. https://doi.org/10.1111/and.13189.

    Article  CAS  PubMed  Google Scholar 

  11. Gat Y, Gornish M, Chakraborty J, Perlow A, Levinger U, Pasqualotto F. Azoospermia and maturation arrest: malfunction of valves in erect poster of humans leads to hypoxia in sperm production site. Andrologia. 2010;42:389–94.

    Article  CAS  Google Scholar 

  12. Cai X, Huang Y, Zhang X, Wang S, Zou Z, Wang G, et al. Cloning, characterization, hypoxia and heat shock response of hypoxia inducible factor-1 (HIF-1) from the small abalone Haliotis diversicolor. Gene. 2014;534(2):256–64. https://doi.org/10.1016/j.gene.2013.10.048.

    Article  CAS  PubMed  Google Scholar 

  13. Semenza GL. Regulation of oxygen homeostasis by hypoxia-inducible factor 1. Physiology (Bethesda). 2009;24:97–106. https://doi.org/10.1152/physiol.00045.2008.

    Article  CAS  Google Scholar 

  14. Brahimi-Horn C, Pouysségur J. The role of the hypoxia-inducible factor in tumor metabolism growth and invasion. Bull Cancer. 2006;93(8):E73–80.

    PubMed  Google Scholar 

  15. Garcia-Maceira P, Mateo J. Silibinin inhibits hypoxia-inducible factor-1alpha and mTOR/p70S6K/4E-BP1 signalling pathway in human cervical and hepatoma cancer cells: implications for anticancer therapy. Oncogene. 2009;28(3):313–24. https://doi.org/10.1038/onc.2008.398.

    Article  CAS  PubMed  Google Scholar 

  16. Kim MH, Jeong YJ, Cho HJ, Hoe HS, Park KK, Park YY, et al. Delphinidin inhibits angiogenesis through the suppression of HIF-1alpha and VEGF expression in A549 lung cancer cells. Oncol Rep. 2017;37(2):777–84. https://doi.org/10.3892/or.2016.5296.

    Article  CAS  PubMed  Google Scholar 

  17. Zhang K, Wang Z, Wang H, Fu Q, Zhang H, Cao Q. Hypoxia-induced apoptosis and mechanism of epididymal dysfunction in rats with left-side varicocele. Andrologia. 2016;48(3):318–24. https://doi.org/10.1111/and.12449.

    Article  CAS  PubMed  Google Scholar 

  18. Goren MR, Kilinc F, Kayaselcuk F, Ozer C, Oguzulgen I, Hasirci E (2017) Effects of experimental left varicocele repair on hypoxia-inducible factor-1alpha and vascular endothelial growth factor expressions and angiogenesis in rat testis. Andrologia. 49(2):e12614. https://doi.org/10.1111/and.12614

  19. Korpelainen EI, Karkkainen MJ, Tenhunen A, Lakso M, Rauvala H, Vierula M, et al. Overexpression of VEGF in testis and epididymis causes infertility in transgenic mice: evidence for nonendothelial targets for VEGF. [Journal Article; Research Support, Non-U.S. Gov’t]. J Cell Biol. 1998;143(6):1705–12. https://doi.org/10.1083/jcb.143.6.1705.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Shiraishi K, Naito K. Involvement of vascular endothelial growth factor on spermatogenesis in testis with varicocele. Fertil Steril. 2008;90(4):1313–6. https://doi.org/10.1016/j.fertnstert.2007.08.030.

    Article  PubMed  Google Scholar 

  21. Xie Y, Shi X, Sheng K, Han G, Li W, Zhao Q, et al. PI3K/Akt signaling transduction pathway, erythropoiesis and glycolysis in hypoxia (review). Mol Med Rep. 2019;19(2):783–91. https://doi.org/10.3892/mmr.2018.9713.

    Article  CAS  PubMed  Google Scholar 

  22. Chambard JC, Lefloch R, Pouyssegur J, Lenormand P. ERK implication in cell cycle regulation. Biochim Biophys Acta. 2007;1773(8):1299–310. https://doi.org/10.1016/j.bbamcr.2006.11.010.

    Article  CAS  PubMed  Google Scholar 

  23. Dery MA, Michaud MD, Richard DE. Hypoxia-inducible factor 1: regulation by hypoxic and non-hypoxic activators. Int J Biochem Cell Biol. 2005;37(3):535–40. https://doi.org/10.1016/j.biocel.2004.08.012.

    Article  CAS  PubMed  Google Scholar 

  24. Marhold M, Tomasich E, El-Gazzar A, Heller G, Spittler A, Horvat R, et al. HIF1alpha regulates mTOR signaling and viability of prostate cancer stem cells. Mol Cancer Res. 2015;13(3):556–64. https://doi.org/10.1158/1541-7786.MCR-14-0153-T.

    Article  CAS  PubMed  Google Scholar 

  25. Park SJ, Ryu J, Kim IH, Choi YH, Nam TJ. Activation of the mTOR signaling pathway in breast cancer MCF7 cells by a peptide derived from Porphyra yezoensis. Oncol Rep. 2015;33(1):19–24. https://doi.org/10.3892/or.2014.3557.

    Article  CAS  PubMed  Google Scholar 

  26. Zhu Q, Wang H, Jiang B, Ni X, Jiang L, Li C, et al. Loss of ATF3 exacerbates liver damage through the activation of mTOR/p70S6K/ HIF-1alpha signaling pathway in liver inflammatory injury. Cell Death Dis. 2018;9(9):910. https://doi.org/10.1038/s41419-018-0894-1.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Zhu SM, Rao T, Yang X, Ning JZ, Yu WM, Ruan Y, et al. Autophagy may play an important role in varicocele. Mol Med Rep. 2017;16(4):5471–9. https://doi.org/10.3892/mmr.2017.7253.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Sengupta P. The laboratory rat: relating its age with human’s. Int J Prev Med. 2013;4(6):624–30.

    PubMed  PubMed Central  Google Scholar 

  29. Turner TT. The study of varicocele through the use of animal models. Hum Reprod Update. 2001;7(1):78–84.

    Article  CAS  Google Scholar 

  30. Cheng S, Zhang X, Feng Q, Chen J, Shen L, Yu P, et al. Astragaloside IV exerts angiogenesis and cardioprotection after myocardial infarction via regulating PTEN/PI3K/Akt signaling pathway. Life Sci. 2019;227:82–93. https://doi.org/10.1016/j.lfs.2019.04.040.

    Article  CAS  PubMed  Google Scholar 

  31. Jin G, Liu J, Qin Q, Gao S, Zhang F, Ma Y, et al. Increased level of c-kit in semen of infertile patients with varicocele. Urol J. 2017;14(2):3023–7.

    PubMed  Google Scholar 

  32. Chen MC, Hsu WL, Hwang PA, Chou TC. Low molecular weight fucoidan inhibits tumor angiogenesis through downregulation of HIF-1/VEGF signaling under hypoxia. Mar Drugs. 2015;13(7):4436–51. https://doi.org/10.3390/md13074436.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Zhu Z, Zhao X, Huang F, Wang F, Wang W. Morinda Officinalis polysaccharides attenuate varicocele-induced spermatogenic impairment through the modulation of angiogenesis and relative factors. Evid Based Complement Alternat Med. 2019;2019:8453635–13. https://doi.org/10.1155/2019/8453635.

    Article  PubMed  PubMed Central  Google Scholar 

  34. Wang H, Wang J, Zhang J, Jin S, Li H. Role of PI3K/AKT/mTOR signaling pathway in DBP-induced apoptosis of testicular sertoli cells in vitro. Environ Toxicol Pharmacol. 2017;53:145–50. https://doi.org/10.1016/j.etap.2017.05.013.

    Article  CAS  PubMed  Google Scholar 

  35. Yuan Y, Wang Y, Hu FF, Jiang CY, Zhang YJ, Yang JL, et al. Cadmium activates reactive oxygen species-dependent AKT/mTOR and mitochondrial apoptotic pathways in neuronal cells. Biomed Environ Sci. 2016;29(2):117–26. https://doi.org/10.3967/bes2016.013.

    Article  CAS  PubMed  Google Scholar 

  36. Wang XQ, Lo CM, Chen L, Ngan ES, Xu A, Poon RY. CDK1-PDK1-PI3K/Akt signaling pathway regulates embryonic and induced pluripotency. Cell Death Differ. 2017;24(1):38–48. https://doi.org/10.1038/cdd.2016.84.

    Article  CAS  PubMed  Google Scholar 

  37. Chen L, Xu B, Liu L, Luo Y, Yin J, Zhou H, et al. Hydrogen peroxide inhibits mTOR signaling by activation of AMPKalpha leading to apoptosis of neuronal cells. Lab Investig. 2010;90(5):762–73. https://doi.org/10.1038/labinvest.2010.36.

    Article  CAS  PubMed  Google Scholar 

  38. Lal BK, Varma S, Pappas PJ, Hobson RW 2nd, Duran WN. VEGF increases permeability of the endothelial cell monolayer by activation of PKB/akt, endothelial nitric-oxide synthase, and MAP kinase pathways. Microvasc Res. 2001;62(3):252–62. https://doi.org/10.1006/mvre.2001.2338.

    Article  CAS  PubMed  Google Scholar 

  39. Chen MC, Lee CF, Huang WH, Chou TC. Magnolol suppresses hypoxia-induced angiogenesis via inhibition of HIF-1alpha/VEGF signaling pathway in human bladder cancer cells. Biochem Pharmacol. 2013;85(9):1278–87. https://doi.org/10.1016/j.bcp.2013.02.009.

    Article  CAS  PubMed  Google Scholar 

  40. Korff T, Augustin HG. Tensional forces in fibrillar extracellular matrices control directional capillary sprouting. J Cell Sci. 1999;112(Pt 19):3249–58.

    CAS  PubMed  Google Scholar 

  41. Shiraishi K, Naito K. Nitric oxide produced in the testis is involved in dilatation of the spermatic vein that compromises spermatogenesis in infertile men with varicocele. BJU Int. 2007;99(5):1086–90. https://doi.org/10.1111/j.1464-410X.2007.06800.x.

    Article  CAS  PubMed  Google Scholar 

  42. Su Y, Quan C, Li X, Shi Y, Duan P, Yang K. Mutual promotion of apoptosis and autophagy in prepubertal rat testes induced by joint exposure of bisphenol A and nonylphenol. Environ Pollut. 2018;243(Pt A):693–702. https://doi.org/10.1016/j.envpol.2018.09.030.

    Article  CAS  PubMed  Google Scholar 

Download references

Funding

This study was supported by the National Natural Science Foundation of China (81401192 and 81973864) and Research Project Supported by Shanxi Scholarship Council of China (2017123).

Author information

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, School of Basic Medicine, Shanxi Medical University, 56 Xinjiannan Road, Taiyuan, 030001, Shanxi, China

    Danfeng Wang & Wei Zhao

  2. Department of Reproductive Medical, Shanxi Provincial People’s Hospital Affiliated to Shanxi Medical University, 29 Shuangtasi Street, Taiyuan, 030012, Shanxi, China

    Jianrong Liu, Caixia Yuan & Qin Qin

  3. Department of Central Laboratory, Shanxi Provincial People’s Hospital Affiliated to Shanxi Medical University, 29 Shuangtasi Street, Taiyuan, 030012, Shanxi, China

    Yimin Wang, Fang Zhang & Guorong Jin

Authors
  1. Danfeng Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wei Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jianrong Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yimin Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Caixia Yuan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Fang Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Guorong Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Qin Qin
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Q.Q.: concept and design, analysis, interpretation of data, revision of the article, and final approval for submission. D.F.W.: design and completion of the experiment, analysis of the results, drafting of the article, revision of the article, and final approval for submission. Y.M.W., F.Z., and J.R.L.: interpretation of data, revision of the article, and final approval for submission. W.Z., C.X.Y., and G.R.J.: discussion of results, drafting of the article, and final approval for submission. All authors have read and approved the final version and submission of this manuscript.

Corresponding author

Correspondence to Qin Qin.

Ethics declarations

Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical Statement

All authors declare that the study was conducted in strict accordance with the rules of good scientific practice and that they are responsible for its contents. All experiments were conducted in a way that can maximize the rigor and reproducibility, without prejudice.

Ethical Approval

All authors declare that this research was done by strictly adhering to the rules of good scientific practice and are responsible for its content. All experiments were performed in a manner that maximized rigor and reproducibility and without bias. All applicable international, national, and/or institutional animal care and use guidelines were followed. All animal experiments were in accordance with the Guide for Care and Use of Laboratory Animals approved by the ethics committee of Shanxi Provincial People’s Hospital (ethics approval number: (2020) provincial medical opinions no. 53).

Consent for Publication

All authors consent to the publication of this manuscript.

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

Wang, D., Zhao, W., Liu, J. et al. Effects of HIF-1α on Spermatogenesis of Varicocele Rats by Regulating VEGF/PI3K/Akt Signaling Pathway. Reprod. Sci. 28, 1161–1174 (2021). https://doi.org/10.1007/s43032-020-00395-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s43032-020-00395-0

Keywords

Search

Navigation