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SIRT5 is associated with asthenozoospermia and regulates GC‑2 spd cell proliferation and apoptosis

  • Authors:
    • Huilin Xing
    • Shu Chen
    • Mingxia Wang
    • Tingting Zheng
    • Bo Yang
  • View Affiliations / Copyright

    Affiliations: Shenzhen Key Laboratory for Drug Addiction and Medication Safety, Institute of Ultrasonic Medicine, Department of Ultrasound, Peking University Shenzhen Hospital, Shenzhen Peking University-Hong Kong University of Science and Technology Medical Center, Shenzhen, Guangdong 518036, P.R. China, The Reproductive Medicine Center, Peking University Shenzhen Hospital, Shenzhen, Guangdong 518036, P.R. China
    Copyright: © Xing et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 269
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    Published online on: July 23, 2025
       https://doi.org/10.3892/mmr.2025.13634
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Abstract

Infertility is a multifactorial condition that affects ~7% of the male population. The mechanism of male infertility primarily involves intrinsic and environmental factors, of which genetic defects are key for its occurrence. Sirtuin 5 (SIRT5) is primarily localized in mitochondria and associated with male reproduction. SIRT5 is downregulated in asthenozoospermic sperm compared with normal sperm. To explore the role of SIRT5 in male reproduction, Cell Counting Kit‑8, EdU, wound healing, and apoptosis assayswere performed in GC‑2 spd cells (mouse spermatocytes) which revealed that SIRT5 inhibited apoptosis and promoted cell proliferation and migration. Western blotting was performed and the results showed that SIRT5 was involved in the proliferation of GC‑2 spd cells by regulating the PI3K/AKT signaling pathway. The present study demonstrated a mechanism of male infertility, which may aid in its treatment.
View Figures

Figure 1

Expression of SIRT5 in human and
mouse sperm. (A) Previous proteomics analysis revealed that SIRT5
expression was significantly decreased in asthenozoospermia
compared with normal sperm (26).
(B) Analysis of the GSE160749 data set revealed that the mRNA
expression of SIRT5 in asthenozoospermia and
asthenoteratozoospermia sperm was decreased compared with normal
sperm. (C) Expression of SIRT5 in asthenozoospermia sperm samples
was lower compared with that in 21 normal samples. (D) SIRT5
protein expression between normal and asthenozoospermia sperm
samples. Immunofluorescent co-localization of SIRT5 and (E) marker
of spermatocytes in meiosis (γH2AX) in murine testes and (F)
mitochondria. Scale bar, 100 µm. (G) SIRT5 expression in testicular
cell species. *P<0.05 vs. N; ##P<0.01 vs. NZS.
SIRT5, sirtuin 5; N, normal; A, asthenozoospermia; NZS,
normozoospermic; AZS, asthenozoospermic; TPM, transcripts per
million.

Figure 2

Knockdown of SIRT5 inhibits
proliferation of GC-2 spd cells. Following transfection with
siRNAs, SIRT5 expression was suppressed in GC-2 spd cells at both
(A) transcriptional and (B) translational levels. After infection
with OE-SIRT5 lentivirus, SIRT5 expression was markedly increased
in GC-2 spd cells at both (C) transcriptional and (D) translational
levels. (E) Viability of GC-2 spd cells treated with siRNAs or
lentivirus. (F) EdU assays revealed the proliferation capacity of
GC-2 spd cells treated with siRNAs or lentivirus. **P<0.01,
***P<0.001 vs. NC; #P<0.05,
###P<0.001 vs. Ctrl. Ctrl, control; si, small
interfering; NC, negative control; OD, optical density; SIRT5,
sirtuin 5; OE, overexpression.

Figure 3

SIRT5 facilitates migration in GC-2
spd cells. (A) Wound healing assay of GC-2 spd cells transfected
with SIRT5 (A) siRNA and (B) OE lentivirus. ***P<0.001 vs. NC;
##P<0.01 vs. Ctrl. SIRT5, sirtuin 5; Ctrl, control;
NC, negative control; siRNA, small interfering RNA; OE,
overexpression.

Figure 4

SIRT5 inhibits apoptosis in GC-2 spd
cells. (A) Proportion of apoptotic SIRT5 (A) knockdown and (B) OE
cells. **P<0.01, ***P<0.001 vs. NC; ##P<0.01
vs. Ctrl. SIRT5, sirtuin 5; Ctrl, control; NC, negative control;
siRNA, small interfering RNA; OE, overexpression.

Figure 5

SIRT5 regulates the PI3K/AKT
signaling pathway in GC-2 spd cells. (A) Western blot analysis of
PI3K, p-AKT, AKT, Bcl-2, Bax and SIRT5 in GC-2 spd cells treated
with siRNA or lentivirus. (B) Semi-quantitative analysis of western
blot results. (C) Co-IP using a PI3K and FLAG antibody on GC-2 spd
cells. *P<0.05, **P<0.01, ***P<0.001 vs. NC;
#P<0.05, ##P<0.01,
###P<0.001 vs. Ctrl. p-, phosphorylated; SIRT5,
sirtuin 5; Ctrl, control; NC, negative control; ns, not
significant; siRNA, small interfering RNA; OE, overexpression; IB,
immunoblot; IP, immunoprecipitation.
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Copy and paste a formatted citation
Spandidos Publications style
Xing H, Chen S, Wang M, Zheng T and Yang B: SIRT5 is associated with asthenozoospermia and regulates GC‑2 spd cell proliferation and apoptosis. Mol Med Rep 32: 269, 2025.
APA
Xing, H., Chen, S., Wang, M., Zheng, T., & Yang, B. (2025). SIRT5 is associated with asthenozoospermia and regulates GC‑2 spd cell proliferation and apoptosis. Molecular Medicine Reports, 32, 269. https://doi.org/10.3892/mmr.2025.13634
MLA
Xing, H., Chen, S., Wang, M., Zheng, T., Yang, B."SIRT5 is associated with asthenozoospermia and regulates GC‑2 spd cell proliferation and apoptosis". Molecular Medicine Reports 32.4 (2025): 269.
Chicago
Xing, H., Chen, S., Wang, M., Zheng, T., Yang, B."SIRT5 is associated with asthenozoospermia and regulates GC‑2 spd cell proliferation and apoptosis". Molecular Medicine Reports 32, no. 4 (2025): 269. https://doi.org/10.3892/mmr.2025.13634
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Spandidos Publications style
Xing H, Chen S, Wang M, Zheng T and Yang B: SIRT5 is associated with asthenozoospermia and regulates GC‑2 spd cell proliferation and apoptosis. Mol Med Rep 32: 269, 2025.
APA
Xing, H., Chen, S., Wang, M., Zheng, T., & Yang, B. (2025). SIRT5 is associated with asthenozoospermia and regulates GC‑2 spd cell proliferation and apoptosis. Molecular Medicine Reports, 32, 269. https://doi.org/10.3892/mmr.2025.13634
MLA
Xing, H., Chen, S., Wang, M., Zheng, T., Yang, B."SIRT5 is associated with asthenozoospermia and regulates GC‑2 spd cell proliferation and apoptosis". Molecular Medicine Reports 32.4 (2025): 269.
Chicago
Xing, H., Chen, S., Wang, M., Zheng, T., Yang, B."SIRT5 is associated with asthenozoospermia and regulates GC‑2 spd cell proliferation and apoptosis". Molecular Medicine Reports 32, no. 4 (2025): 269. https://doi.org/10.3892/mmr.2025.13634
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