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Volume 27
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10.3390/ijms27156817
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Dose-Dependent Effects of Protocatechuic Acid on Motility, Redox Balance, and DNA Integrity of Frozen–Thawed Ram Spermatozoa

by
Umut Taşdemir
1,*,
Fatih Avdatek
2,
Muhammed Enes İnanç
3,
Şükrü Güngör
3,
Mehmet Fuat Gülhan
4,
Barış Denk
5,
Murat Kırıkkulak
2 and
Deniz Yeni
2
1
Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Ankara University, Ankara 06070, Türkiye
2
Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Afyon Kocatepe University, Afyonkarahisar 03200, Türkiye
3
Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Burdur Mehmet Akif Ersoy University, Burdur 15030, Türkiye
4
Department of Medicinal Aromatic Plants, Technical Sciences Vocational School, Aksaray University, Aksaray 68100, Türkiye
5
Department of Biochemistry, Faculty of Veterinary Medicine, Afyon Kocatepe University, Afyonkarahisar 03200, Türkiye
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2026, 27(15), 6817; https://doi.org/10.3390/ijms27156817
Submission received: 30 June 2026 / Revised: 18 July 2026 / Accepted: 28 July 2026 / Published: 29 July 2026
Versions Notes

Abstract

This study investigated the effects of supplementing Tris semen extender with protocatechuic acid (PCA) at doses of 25, 50, and 100 µg/mL on the quality of ram spermatozoa after cryopreservation. We evaluated motility, kinematic parameters, DNA integrity, redox status, plasma membrane–acrosome integrity (PMAI), viability, and high mitochondrial membrane potential (HMMP). PCA supplementation produced concentration-dependent and parameter-specific responses. The 25 µg/mL PCA group exhibited the most favorable functional profile, demonstrating improved progressive and total motility alongside reduced DNA damage. This protective effect was evidenced by lower tail length, tail DNA percentage, and tail moment in the comet assay. Conversely, treatment with 100 µg/mL PCA appeared to exert a notable effect on the redox balance, indicated by lower malondialdehyde, total oxidant status, and oxidative stress index levels, alongside an elevated total antioxidant status (p < 0.05). However, this higher concentration (100 µg/mL) did not provide parallel DNA protection and was associated with increased comet assay indices. PMAI, viability, and HMMP were not significantly affected by PCA supplementation (p > 0.05). These findings indicate that PCA exerts dose-dependent but parameter-specific effects on frozen–thawed ram spermatozoa. Low-dose PCA, particularly 25 µg/mL, appears more suitable for preserving motility and nuclear integrity, whereas 100 µg/mL mainly improves oxidative balance.
Keywords: protocatechuic acid; ram semen; cryopreservation; oxidative stress; DNA integrity; sperm motility; antioxidant protocatechuic acid; ram semen; cryopreservation; oxidative stress; DNA integrity; sperm motility; antioxidant

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MDPI and ACS Style

Taşdemir, U.; Avdatek, F.; İnanç, M.E.; Güngör, Ş.; Gülhan, M.F.; Denk, B.; Kırıkkulak, M.; Yeni, D. Dose-Dependent Effects of Protocatechuic Acid on Motility, Redox Balance, and DNA Integrity of Frozen–Thawed Ram Spermatozoa. Int. J. Mol. Sci. 2026, 27, 6817. https://doi.org/10.3390/ijms27156817

AMA Style

Taşdemir U, Avdatek F, İnanç ME, Güngör Ş, Gülhan MF, Denk B, Kırıkkulak M, Yeni D. Dose-Dependent Effects of Protocatechuic Acid on Motility, Redox Balance, and DNA Integrity of Frozen–Thawed Ram Spermatozoa. International Journal of Molecular Sciences. 2026; 27(15):6817. https://doi.org/10.3390/ijms27156817

Chicago/Turabian Style

Taşdemir, Umut, Fatih Avdatek, Muhammed Enes İnanç, Şükrü Güngör, Mehmet Fuat Gülhan, Barış Denk, Murat Kırıkkulak, and Deniz Yeni. 2026. "Dose-Dependent Effects of Protocatechuic Acid on Motility, Redox Balance, and DNA Integrity of Frozen–Thawed Ram Spermatozoa" International Journal of Molecular Sciences 27, no. 15: 6817. https://doi.org/10.3390/ijms27156817

APA Style

Taşdemir, U., Avdatek, F., İnanç, M. E., Güngör, Ş., Gülhan, M. F., Denk, B., Kırıkkulak, M., & Yeni, D. (2026). Dose-Dependent Effects of Protocatechuic Acid on Motility, Redox Balance, and DNA Integrity of Frozen–Thawed Ram Spermatozoa. International Journal of Molecular Sciences, 27(15), 6817. https://doi.org/10.3390/ijms27156817

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