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The Beneficial Impact of Pomegranate Oil Nanoemulsion on the Quality of Cryopreserved Bovine Sperm: Antioxidant and Anti-Apoptotic Effects. |
Fatma Mohsen Shalaby1 
, Soha A. Hassan2
, Ali Ali El-Raghi3,*
, Fatemah Enad Alajmi4
, Mohamed G. Alfayoumi5
, Kandil Abd El Hai Attia6
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1Department of Biology, Faculty of Sciences, King Khalid University, Abha 62521, KSA
2Department of Biotechnology, Faculty of Applied Health Sciences, October 6 University, Giza, Egypt
3Department of Animal, Poultry and Fish Production, Faculty of Agriculture, Damietta University, 34517, Damietta, Egypt
4Department of Biology, College of Science, University of Hafr Al Batin, Hafr Al Batin 39524, Saudi Arabia
5Toxicology Expert, Gen. Dept. of Forensic Science and Criminology, Dubai Police General H.Q
6Evaluation of Natural Resources Department, Environmental Studies and Research Institute, University of Sadat City, Minufiya 32897, Egypt |
Correspondence:
Ali Ali El-Raghi,Email: ali21384@yahoo.com
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Received: 4 February 2025 • Revised: 15 March 2025 • Accepted: 7 April 2025 |
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| Abstract |
Objective
This study was conducted to investigate the effect of Pomegranate oil nano-emulsion (PO-NE) supplementation in semen extender on semen quality, redox status and apoptotic genes of cryopreserved bovine semen.
Methods
Seven proven fertility Holstein cattle bulls (4–6 years) were involved, and the semen samples were collected using the artificial vagina method. Semen was pooled and cryopreserved in tris extender containing PO-NE at 0 (PO-NE0), 1 (PO-NE1), 2 (PO-NE 2) and 4 μg/mL (PO-NE4), respectively.
Results
Incorporating 2 or 4 μg/mL PO-NE into freezing media significantly increased sperm progressive motility, viability, membrane integrity, and kinematic parameters. Furthermore, the aforementioned two treated concentrations demonstrated superior anti-oxidative activities (total antioxidant capacity and super oxide dismutase) and higher nitric oxide levels compared to the control group (p < 0.05). The levels of hydrogen peroxide, malondialdehyde, and Nuclear Factor-Kappa B were notably lower in the PO-NE4 treated group compared to the control group. The addition of 2 or 4 μg/mL of PO-NE to the freezing media significantly downregulated pro-apoptotic genes (caspase 3, Bax), while significantly induced the expression of anti-apoptotic gene Bcl2. The addition of PO-NE preserved plasma membrane and acrosome integrity and maintained the ultrastructure of sperm, contrasting with PO-NE0, which exhibited the most damage.
Conclusion
supplementing the bovine freezing extender with 2 - 4 μg/mL of PO-NE enhanced post-thawed sperm characteristics by reducing oxidative stress, improving antioxidant indices and apoptotic genes expression, and preserving the ultrastructure integrity of frozen-thawed cattle sperm.
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| Keywords:
Antioxidant; Apoptotic genes; Bovine semen cryopreservation; Pomegranate Oil Nanoemulsion; Ultrastructure |
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