Protective effects of hesperetin on the quality of sperm, apoptosis, lipid peroxidation, and oxidative stress during the process of cryopreservation: An experimental study

Abstract

Background: Hesperetin is a bioflavonoid compound, largely used in Chinese traditional medicine and found plenty in citrus fruits. Hesperetin has beneficial effects against different diseases. The sperm cryopreservation process is a common method that is used in infertility laboratories. It has been reported that during the cryopreservation process, the quality of sperm is significantly reduced.
 Objective: To investigate the effect of hesperetin on the quality of human spermatozoa during the cryopreservation process.
 Materials and Methods: In this experimental study, 22 sperm sample of normozoospermia men who referred to the infertility department of the Shariati Hospital (Tehran, Iran) Between October and November 2019 were collect and divided into three groups as: 1) fresh, 2) control (frozen-thawed group without treatment), and 3) treatment group as frozen-thawed samples supplemented with 20 μM hesperetin. Motility, Viability, morphology, Apoptotic-like changes, intracellular H2O2, intracellular O2−, and lipid peroxidation (LPO) was measured.
 Results: Hesperetin treatment during the cryopreservation process of human sperm significantly improved the viability, motility, and morphology rates of the spermatozoa after frozen-thawed process in control group (p < 0.01). In addition, it significantly reduced the reactive oxygen species (ROS) level, LPO level and increased the percentage of viable sperm cells with intact plasma membrane (p < 0.01) after frozen-thawed process.
 Conclusion: Hesperetin can improve the quality of human sperm and also protect human sperm against reactive oxygen species, LPO, and apoptosis during the cryopreservation-thawing process.
 Key words: Cryopreservation, Hesperetin, Spermatozoa, Reactive oxygen species.

Investigation of Changes in Spermatozoa Characteristics, Chromatin Structure, and Antioxidant/Oxidant Parameters after Freeze-Thawing of Hesperidin (Vitamin P) Doses Added to Ram Semen

“…Contrary to our findings, [U]. Valipour et al [28]. showed that the level of ROS formation increased during the freeze-thawing process, and 20 μM HSP treatment significantly reduced ROS levels.…”

“…Samie et al [27] reported that the dose of HSP (50 mg/kg/day) administered to rats contributed to an increase in sperm counts and motility. In another study, 20 µM hesperidin treatment during the cryopreservation of human sperm significantly improved the motility and morphology ratios of spermatozoa after cryopreservation [28]. Aksu et al [29] also showed that the application of HSP (300 mg/kg/day) for 7 days against reproductive damage in male rats caused an increase in the percentage of total sperm motility.…”

“…İsmail et al [38] reported that curcumin nanoparticles (100 μg) added to goat semen extender preserved sperm viability and plasma membrane integrity. In another study, Valipour et al [28] found that 20 µM HSP treatment during the cryopreservation of human sperm significantly improved the viability of the sperm after thawing and reduced LPL. The results of our study showed that HSP kept the viability of spermatozoa stable after cryopreservation in the HSP10, HSP50, and HSP100 dose groups compared to C, but it reduced viability at the HSP250 and HSP500 doses.…”

“…The HSP10 and HSP50 doses can prevent the formation of ROS and contribute to the protection of polyunsaturated oxidized fatty acids in sperm against oxidation, thus increasing the efficiency of living cells and fertilization. Additionally, the ability of HPS to stabilize the mitochondrial membrane was reported to protect sperm cells from apoptosis and increase mitochondrial functions [28]. There is some evidence that HSP has a protective effect against oxidative and nitrosative modifications in DNA induced by endogenous and exogenous ROS production [27,44,47].…”

Evaluation of the effects of hesperidin on fresh and frozen-thawed semen quality using two different cryopreservation methods in Simmental bull

“…Then, 100 µL of the stock solution was mixed with 900 µL distilled water (100 µM Hesp solution). Finally, 10 and 20 µL of 100 µM Hesp solution were dissolved with 90 and 80 µL pre-extender sperm solution, respectively, and 1 and 2 µM Hesp solutions were prepared (Ali et al, 2021;Valipour et al, 2021). Hesp supplementation was performed according to the one-and two-step dilution protocols separately.…”