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Effect of silencing HIF-1α gene on testicle spermatogenesis function in varicocele rats

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Abstract

This study uses the CRISPR/Cas9 gene editing technique to silence the expression of hypoxia-inducible factor-1α (HIF-1α) gene and investigate its effect on testicle spermatogenesis function in varicocele (VC) rats. Sprague Dawley rats were divided into four groups; the control, VC model, VC+HIF-1α-lentivirus and VC+Luciferase-lentivirus group. The sperm count and survival rate were analyzed using computer-aided sperm analysis. The morphological changes of seminiferous tubules were observed by a microscope. Expressions of HIF-1α, Bax, cleaved caspase-3 and Bcl-2 were detected via Western blot, immunofluorescence and real-time polymerase chain reaction methods. One-way ANOVA was used to analyze the differences between groups. The sperm count and survival rate were significantly lower (p < 0.05) and the seminiferous epithelium was more disordered in the VC group than that in the control group. The expression of Bax and cleaved caspase-3 were increased and Bcl-2 was reduced in the VC group than the control group. Compared with the VC group, sperm count and survival rate noticeably increased (p < 0.05), seminiferous epithelium was inordered arrangement and fewer spermatogenic cells were injured in the VC+HIF-1α-lentivirus group. Expression of Bax and cleaved caspase-3 were decreased significantly in the VC+HIF-1α-lentivirus group compared with the VC group and VC+Luciferase-lentivirus group (p < 0.05), whereas the expression of Bcl-2 was increased (p < 0.05). No significant difference was observed between the control group and the VC+HIF-1α-lentivirus group (p > 0.05). Results show that the apoptosis of spermatogenic cells was decreased and the testicle spermatogenesis function was significantly improved after silencing HIF-1α gene in testis of VC rats. HIF-1α may play a crucial role during spermatogenesis in VC inducing male infertility.

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Funding

Supported by the National Natural Science Foundation of China (81401192), Research Project Supported by Shanxi Scholarship Council of China (2017123) and Fund Program for the Scientific Activities of Selected Returned Overseas Professionals in Shanxi Province.

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Authors and Affiliations

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

    Wei Zhao, Danfeng Wang & Qin Qin

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

    Jianrong Liu, Caixia Yuan, Xin Wang & Qin Qin

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

    Yimin Wang, Fang Zhang & Guorong Jin

Authors
  1. Wei Zhao
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  2. Jianrong Liu
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  3. Danfeng Wang
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  4. Yimin Wang
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  5. Fang Zhang
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  7. Caixia Yuan
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  8. Xin Wang
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  9. Qin Qin
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Correspondence to Qin Qin.

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Ethical statement

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.

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All applicable international, national and/or institutional guidelines for the care and use of animals were followed. All animal experiments conformed with the Guide for Care and Use of Laboratory Animals and were approved by Shanxi Provincial People’s Hospital.

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Zhao, W., Liu, J., Wang, D. et al. Effect of silencing HIF-1α gene on testicle spermatogenesis function in varicocele rats. Cell Tissue Res 378, 543–554 (2019). https://doi.org/10.1007/s00441-019-03064-0

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