Ameliorative Impacts of Carvacrol on DNA Integrity, Oxidative Stress, and Sperm Quality in Asthenozoospermic Infertile Individuals During Cryopreservation

Overview

Journal Naunyn Schmiedebergs Arch Pharmacol

Specialty Pharmacology

Date 2024 Nov 16

PMID 39549060

Authors

Ebrahim Cheraghi

Seyed Mohammad Ali Shariatzadeh

Fatemeh Hajiazimi

Affiliations

Soon will be listed here.

Abstract

Nowadays, the cryopreservation process can produce reactive oxygen species (ROS), which cause damage to the motility, cell membrane, and DNA integrity of sperm. This study is aimed at evaluating the impact of carvacrol, as a powerful antioxidant, on sperm features and improving oxidative stress throughout the cryopreservation procedure. In this prospective study, semen samples from 25 patients with asthenozoospermia were separated into three groups: fresh, freezing, and freezing + carvacrol (a dose of 100 µM). The subsequent parameters were evaluated using standard methods in all three groups: sperm motility according to WHO criteria, sperm morphology using Papanicolaou staining, sperm viability with eosin-nigrosin staining, DNA integrity with acridine orange staining, levels of antioxidant enzymes (catalase, glutathione, and superoxide dismutase), total antioxidant capacity (TAC), and malondialdehyde (MDA) using ELISA. Also, DNA fragmentation was analyzed by the SDFA kit, and mitochondrial membrane potential (MMP) was assessed by rhodamine staining. Average sperm viability, motility, mitochondrial membrane potentiality, integrity of sperm membrane, and antioxidant enzyme levels are meaningfully reduced in the freezing group in contrast to the control group. The freezing group showed a meaningful rise in the mean MDA levels and DNA fragmentation compared to the control group. In the freezing group supplemented with carvacrol, a meaningful rise could be visible in mean percentages of viability, motility, and antioxidant enzyme levels, whereas mean levels of MDA and DNA fragmentation meaningfully declined in contrast to the freezing group. The results demonstrate that carvacrol, a potent antioxidant, offers significant protection against the loss generated by the freeze-thaw procedure, thereby improving sperm quality.

References

Agarwal A, Saleh R, Bedaiwy M . Role of reactive oxygen species in the pathophysiology of human reproduction. Fertil Steril. 2003; 79(4):829-43. DOI: 10.1016/s0015-0282(02)04948-8. View

Agharezaee N, Hashemi M, Shahani M, Gilany K . Male Infertility, Precision Medicine and Systems Proteomics. J Reprod Infertil. 2019; 19(4):185-192. PMC: 6328981. View

Anger J, Gilbert B, Goldstein M . Cryopreservation of sperm: indications, methods and results. J Urol. 2003; 170(4 Pt 1):1079-84. DOI: 10.1097/01.ju.0000084820.98430.b8. View

Asa E, Ahmadi R, Mahmoodi M, Mohammadniya A . Supplementation of freezing media with alpha lipoic acid preserves the structural and functional characteristics of sperm against cryodamage in infertile men with asthenoteratozoospermia. Cryobiology. 2020; 96:166-174. DOI: 10.1016/j.cryobiol.2020.07.001. View

Ashrafi I, Kohram H, Ardabili F . Antioxidative effects of melatonin on kinetics, microscopic and oxidative parameters of cryopreserved bull spermatozoa. Anim Reprod Sci. 2013; 139(1-4):25-30. DOI: 10.1016/j.anireprosci.2013.03.016. View

Azizi M, Cheraghi E, Mehranjani M . Effect of Myo-inositol on sperm quality and biochemical factors in cryopreserved semen of patients with Asthenospermia. Andrologia. 2022; 54(10):e14528. DOI: 10.1111/and.14528. View

Cheraghi E, Sajadi S, Mehranjani M . The effect of Quercetin on the quality of sperm parameters in frozen-thawed semen of patients with Asthenospermia. Andrologia. 2021; 53(9):e14167. DOI: 10.1111/and.14167. View

Chikhoune A, Stouvenel L, Iguer-Ouada M, Hazzit M, Schmitt A, Lores P . In-vitro effects of Thymus munbyanus essential oil and thymol on human sperm motility and function. Reprod Biomed Online. 2015; 31(3):411-20. DOI: 10.1016/j.rbmo.2015.06.011. View

Di Santo M, Tarozzi N, Nadalini M, Borini A . Human Sperm Cryopreservation: Update on Techniques, Effect on DNA Integrity, and Implications for ART. Adv Urol. 2011; 2012:854837. PMC: 3238352. DOI: 10.1155/2012/854837. View

10.

El-Nekeety A, Mohamed S, Hathout A, Hassan N, Aly S, Abdel-Wahhab M . Antioxidant properties of Thymus vulgaris oil against aflatoxin-induce oxidative stress in male rats. Toxicon. 2011; 57(7-8):984-91. DOI: 10.1016/j.toxicon.2011.03.021. View

11.

Fernandez J, Muriel L, Goyanes V, Segrelles E, Gosalvez J, Enciso M . Simple determination of human sperm DNA fragmentation with an improved sperm chromatin dispersion test. Fertil Steril. 2005; 84(4):833-42. DOI: 10.1016/j.fertnstert.2004.11.089. View

12.

Guvenc M, Cellat M, Gokcek I, Yavas I, Ozsoy S . Effects of thymol and carvacrol on sperm quality and oxidant/antioxidant balance in rats. Arch Physiol Biochem. 2018; 125(5):396-403. DOI: 10.1080/13813455.2018.1476979. View

13.

Hamada A, Montgomery B, Agarwal A . Male infertility: a critical review of pharmacologic management. Expert Opin Pharmacother. 2012; 13(17):2511-31. DOI: 10.1517/14656566.2012.740011. View

14.

Hezavehei M, Sharafi M, Mohseni Kouchesfahani H, Henkel R, Agarwal A, Esmaeili V . Sperm cryopreservation: A review on current molecular cryobiology and advanced approaches. Reprod Biomed Online. 2018; 37(3):327-339. DOI: 10.1016/j.rbmo.2018.05.012. View

15.

Irvine D . Epidemiology and aetiology of male infertility. Hum Reprod. 1998; 13 Suppl 1:33-44. DOI: 10.1093/humrep/13.suppl_1.33. View

16.

Ismail A, Abdel-Khalek A, Khalil W, Yousif A, Saadeldin I, Abomughaid M . Effects of mint, thyme, and curcumin extract nanoformulations on the sperm quality, apoptosis, chromatin decondensation, enzyme activity, and oxidative status of cryopreserved goat semen. Cryobiology. 2020; 97:144-152. DOI: 10.1016/j.cryobiol.2020.09.002. View

17.

Kchikich A, Kirschvink N, Raes M, El Otmani S, Chebli Y, Bister J . Carvacrol and Thymol Enhance the Quality of Beni Arouss Buck Semen Stored at 4 °C Thanks to Their Antimicrobial Properties. Vet Sci. 2024; 11(9). PMC: 11435801. DOI: 10.3390/vetsci11090406. View

18.

Kohzadi R, Cheraghi E, Mehranjani M, Shariatzadeh M . Sildenafil citrate ameliorates the adverse effects of cryopreservation on sperm quality in asthenozoospermic men. Cryobiology. 2023; 111:126-133. DOI: 10.1016/j.cryobiol.2023.05.005. View

19.

Ramu S, Jeyendran R . The hypo-osmotic swelling test for evaluation of sperm membrane integrity. Methods Mol Biol. 2012; 927:21-5. DOI: 10.1007/978-1-62703-038-0_3. View

20.

Rozati H, Handley T, Jayasena C . Process and Pitfalls of Sperm Cryopreservation. J Clin Med. 2017; 6(9). PMC: 5615282. DOI: 10.3390/jcm6090089. View