• Clinical and Experimental Reproductive Medicine
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• v.51 no.2
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• pp.102-111
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• 2024

Objective: Given the noteworthy implications of alcohol consumption and its association with male infertility, there has been a notable focus on investigating natural alternatives to mitigate its adverse effects. Thus, this study was conducted to assess the potential protective effect of phycocyanin extract derived from the blue algae Arthrospira (Spirulina) platensis against ethanol-induced oxidative stress, disturbances in testicular morphology, and alterations in sperm production. Methods: Male rats were divided into four groups (five rats each): the control group received a saline solution, the ethanol exposed group (EtOH) was subjected to intraperitoneal injections of 10 mL/kg of ethanol solution at a concentration of 38% (v/v), the phycocyanin alone treated group (P) received oral administration of phycocyanin at a dosage of 50 mg/kg, and the phycocyanin-cotreated group (PE) was given oral phycocyanin followed by ethanol injections. All treatments were administered over a period of 14 days. Results: Our findings demonstrated that ethanol exposure induced reproductive toxicity, characterized by reduced sperm production and viability, alterations in testicular weight and morphology, increased lipid peroxidation levels, and elevated oxidative enzyme activity. In addition, the ethanol-intoxicated group showed perturbations in serum biochemical parameters. However, the simultaneous exposure to ethanol and phycocyanin exhibited a counteractive effect against ethanol toxicity. Conclusion: The results showed that supplementation of phycocyanin prevented oxidative and testicular morphological damage-induced by ethanol and maintained normal sperm production, and viability.

• Journal of Animal Reproduction and Biotechnology
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• v.39 no.2
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• pp.67-80
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• 2024

Background: Post-ovulatory aging (POA) of oocytes is related to a decrease in the quality and quantity of oocytes caused by aging. Previous studies on the characteristics of POA have investigated injury to early embryonic developmental ability, but no information is available on its effects on mitochondrial fission and mitophagy-related responses. In this study, we aimed to elucidate the molecular mechanisms underlying mitochondrial fission and mitophagy in in vitro maturation (IVM) oocytes and a POA model based on RNA sequencing analysis. Methods: The POA model was obtained through an additional 24 h culture following the IVM of matured oocytes. NMN treatment was administered at a concentration of 25 μM during the oocyte culture process. We conducted MitoTracker staining and Western blot experiments to confirm changes in mitochondrial function between the IVM and POA groups. Additionally, comparative transcriptome analysis was performed to identify differentially expressed genes and associated changes in mitochondrial dynamics between porcine IVM and POA model oocytes. Results: In total, 32 common genes of apoptosis and 42 mitochondrial fission and function uniquely expressed genes were detected (≥ 1.5-fold change) in POA and porcine metaphase II oocytes, respectively. Functional analyses of mitochondrial fission, oxidative stress, mitophagy, autophagy, and cellular apoptosis were observed as the major changes in regulated biological processes for oocyte quality and maturation ability compared with the POA model. Additionally, we revealed that the activation of NAD⁺ by nicotinamide mononucleotide not only partly improved oocyte quality but also mitochondrial fission and mitophagy activation in the POA porcine model. Conclusions: In summary, our data indicate that mitochondrial fission and function play roles in controlling oxidative stress, mitophagy, and apoptosis during maturation in POA porcine oocytes. Additionally, we found that NAD⁺ biosynthesis is an important pathway that mediates the effects of DRP1-derived mitochondrial morphology, dynamic balance, and mitophagy in the POA model.

• The Korean Journal of Physiology and Pharmacology
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• v.7 no.6
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• pp.325-331
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• 2003

3-Nitropropionic acid (3-NP) inhibits electron transport in mitochondria, leading to a metabolic failure. In order to elucidate the mechanism underlying this toxicity, we examined a few biochemical changes possibly involved in the process, such as metabolic inhibition, generation of reactive oxygen species (ROS), DNA strand breakage, and activation of Poly(ADP-ribose) polymerase (PARP). Exposure of SK-N-BE(2)C neuroblastoma cells to 3-NP for 48 h caused actual cell death, while inhibition of mitochondrial function was readily observed when exposed for 24 h to low concentrations (0.2${\sim}$2 mM) of 3-NP. The earliest biochemical change detected with low concentration of 3-NP was an accumulation of ROS (4 h after 3-NP exposure) followed by degradation of DNA. PARP activation by damaged DNA was also detectable, but at a later time. The accumulation of ROS and DNA strand breakage were suppressed by the addition of glutathione or N-acetyl-L-cysteine (NAC), which also partially restored mitochondrial function and cell viability. In addition, inhibition of PARP also reduced the 3-NP-induced DNA strand breakage and cytotoxicity. These results suggest that oxidative stress and activation of PARP are the major factors in 3-NP-induced cytotoxicity, and that the inhibition of these factors may be useful in protecting neuroblastoma cells from 3-NP-induced toxicity.

• Development and Reproduction
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• v.10 no.2
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• pp.105-113
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• 2006

Reactive oxygen species(ROS) generated in cellular metabolism have an effect on cell maturation and development. In human reproductive tract, oxidative injury by ROS may induce female infertility. Also, oxidative injury may be responsible for developmental retardation and arrest of mammalian preimplantation embryos. Activating transcription factor 4(ATF4) is a member of the cyclic-AMP response element-binding(CREB) familiy of basic region- leucine zipper(bZip). ATF4 is known to regulate stress response to protect cell from various stress factors and inducer of apoptisis. The purpose of this study was to investigate whether ATF4 is involved in the defensive mechanism in oxidative stress condition during the development of mouse preimplantation embryos. To verify the expression of ATF4 in oxidative stress condition, 2-cell stage embryos were cultured in HTF media containing 0.1mM, 0.5mM or 1mM hydrogen peroxide($H_2O_2$) for 1hr(2-cell), 8hr(4-cell), 17hr(8-cell), 24hr(morula), 48hr(early blastocyst) or 64hr(late blastocyst). The developmental rate decreased in the 0.1mM $H_2O_2$ treated group compared with control group. In embryos treated with 0.5mM and 1mM $H_2O_2$ showed 2-cell block. As a results of the semi-quantitative RT-PCR analysis of SOD1, ATF4 and Bax gene expression, SOD1, ATF4 and Bax genes were increased in 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. In 2-cell embryos, expression of SOD1, ATF4 and Bax genes were notably increased in 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. Immunofluorescence analysis showed that ATF4 protein was localized at the cytoplasm of preimplantation embryos. The increase in ATF4 immunoreactivety was observed in the 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. It suggests that oxidative stress by $H_2O_2$ induces expression of ATF4 and may be involved in protection mechanism in preimplantation embryos from oxidative injury.

• Journal of Animal Reproduction and Biotechnology
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• v.36 no.4
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• pp.261-269
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• 2021

Because sows are industrially vital for swine production, monitoring for their health or disorder status is important to ensure high reproductive performance. Especially, ambient temperature changes in different season, especially during summer, are directly influenced to the reproductive performance of sows. Although the serum biochemical parameters are widely applied in the veterinary medicine with wide ranges for the physiological process, the values are also influenced by several factors such as age, breed, gender, and stress. In addition, domestic sows in Korea-specific reference interval (RI) for serum biochemistry has not been established yet. Therefore, the present study was aimed to evaluate seasonal variation of RIs in the serum biochemistry in domestic sows in Korea at different seasons and to establish normal RIs using a RI finding program (Reference Value Advisor). Significant difference (p < 0.05) on the different seasons were identified in several serum biochemical parameters including BUN, CRE, GGT, GLU, ALB, TP, LDH and Na in sows. Therefore, we further established RIs, specific in domestic sows in Korea regardless of season. The established RIs based on the serum biochemical values provide a baseline for interpreting biochemical results in the domestic sows in Korea, regardless of seasonal effect. It may contribute to develop a strategy for better reproductive performance by improving breeding management practice and evaluating health of pig herds, which facilitate to avert the economic loss in summer infertility in sows.

• Clinical and Experimental Reproductive Medicine
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• v.50 no.4
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• pp.230-243
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• 2023

Objective: High temperatures can trigger cellular oxidative stress and disrupt spermatogenesis, potentially leading to male infertility. We investigated the effects of retinoic acid (RA), chitosan nanoparticles (CHNPs), and retinoic acid loaded with chitosan nanoparticles (RACHNPs) on spermatogenesis in mice induced by scrotal hyperthermia (Hyp). Methods: Thirty mice (weighing 25 to 30 g) were divided into five experimental groups of six mice each. The groups were as follows: control, Hyp induced by a water bath (43 ℃C for 30 minutes/day for 5 weeks), Hyp+RA (2 mg/kg/day), Hyp+CHNPs (2 mg/kg/72 hours), and Hyp+RACHNPs (4 mg/kg/72 hours). The mice were treated for 35 days. After the experimental treatments, the animals were euthanized. Sperm samples were collected for analysis of sperm parameters, and blood serum was isolated for testosterone measurement. Testis samples were also collected for histopathology assessment, reactive oxygen species (ROS) evaluation, and RNA extraction, which was done to compare the expression levels of the bax, bcl2, p53, Fas, and FasL genes among groups. Additionally, immunohistochemical staining was performed. Results: Treatment with RACHNPs significantly increased stereological parameters such as testicular volume, seminiferous tubule length, and testicular cell count. Additionally, it increased testosterone concentration and improved sperm parameters. We observed significant decreases in ROS production and caspase-3 immunostaining in the RACHNP group. Moreover, the expression levels of bax, p53, Fas, and FasL significantly decreased in the groups treated with RACHNPs and RA. Conclusion: RACHNPs can be considered a potent antioxidative and antiapoptotic agent for therapeutic strategies in reproductive and regenerative medicine.

• The Korea Journal of Herbology
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• v.31 no.6
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• pp.1-9
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• 2016

Objectives : Gamisoyosan (GMS) is a useful prescription for treating insomnia, dysmenorrhea and infertility induced by a stress. Also, GMS has been used traditionally to improve systemic circulation and biological energy production. The purpose of this study was to assess the anti-oxidant activity and anti-inflammatory effects of Gamisoyosan Formulation (Soft extract, GMS-SE). Methods : The biological activities such as anti-oxidant and anti-inflammatory effects were measured through cell line-based in vitro assay. We investigated the anti-oxidant properties of GMS-SE on the 1,1-diphenyl-2-picryhydrazyl (DPPH) radical, contents of total flavonoid and polyphenol. GMS-SE compared to butyl hydroxy anizole (BHA). Furthermore, based on this result the anti-inflammatory effects of GMS-SE have verified by mechanism from LPS- treated Raw264.7 macrophages. Results : The anti-oxidant activities of GMS-SE increased markedly, in a dose-dependent manner. The GMS-SE showed significant scavenging activity (GMS-SE $500{\mu}g/m{\ell}$ : $32.77{\pm}1.65%$, GMS-SE $1000{\mu}g/m{\ell}$ : $45.06{\pm}1.04%$ and BHA $100{\mu}g/m{\ell}$ : $39.25{\pm}2.41%$ for DPPH assay). and, The total phenolic compound and flavonoids contents of GMS-SE were $73.93{\pm}6.87{\mu}g/mg$ and $698.75{\pm}6.78{\mu}g/mg$. GMS-SE which is LPS has diminished in the LPS-induced release of inflammatory mediators (NO, iNOS, COX2 and PGE2) and pro-inflammatory cytokines (TNF-${\alpha}$, IL-6 and IL-$1{\beta}$) from the RAW264.7 macrophages. Moreover, GMS-SE inhibited the activation of phosphorylation of p38 and ERK MAPKs by induced LPS. Conclusion : The present results indicate that GMS-SE has an anti-oxidant and anti-inflammatory properties, therefore may be beneficial in diseases which related to oxidative stress-mediated inflammatory disorders.

• Clinical and Experimental Reproductive Medicine
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• v.34 no.3
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• pp.159-166
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• 2007

Objective: To investigate whether semen parameters in infertile couples who undergone intrauterine insemination (IUI) change in the subsequent IUI cycle and the subsequent in vitro fertilization (IVF) cycle. Methods: Fifty-three infertile couples who had failed to become pregnant after the first IUI cycle with computer-assisted semen analysis (CASA) were included. After the first IUI, thirty-eight couples underwent the second IUI (Group 1), and fifteen underwent IVF-ET procedure (Group 2). All semen parameters including semen volume, concentration, motility and total motile sperm count were analyzed in the second IUI or IVF-ET procedure for comparison with the result of first IUI. Results: There were no significant differences in husband age, interval between the first and second procedure and cause of infertility. In Group 1, only sperm motility at the time of the latter IUI was significantly decreased when compared to the former IUI irrespective of the first semen parameters. In Group 2, sperm concentration, motility and total motile sperm count at the time of subsequent IVF were lower than the former IUI. By sub-analyses of Group 2, in the group of optimal semen parameter at IUI cycle, sperm concentration and total motile sperm count at the time of subsequent IVF were lower than the former IUI, while in the group of suboptimal semen parameter at IUI cycle, only sperm motility at the time of subsequent IVF were lower than the former IUI. Conclusion: The semen parameters in couples converted to IVF cycle were more adversely affected than those remained in IUI cycle. Further study on psychological stress should be necessary to explain the reason.

• Reproductive and Developmental Biology
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• v.35 no.4
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• pp.441-447
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• 2011

The study aim is to investigate the free radicals scavenging and spermatogenic potentials, as well as to analyze any reproductive toxicity of ethanolic extract of Mucuna prureins (M. pruriens) Linn. in spermatozoa, under different dosages in normal male rat. Normal rats were randomly selected and suspension of the extract was administered orally at the dosages of 150, 200 and 250 mg/kg body weight of the different groups of male rats (n=6) once in a day for 60 days and grouped as group II, III and IV respectively. Saline treated rats served as control -group I. On the $60^{th}$ day the animals were sacrificed and the epididymal sperm were subjected to various analyses like level of ROS production, LPO, enzymatic and non enzymatic antioxidant, morphology, morphometry, chromosomal integrity and DNA damage. Results showed significant reduction in ROS production and peroxidation and significant increase in both enzymic and non-enzymic antioxidants in all concentration treated groups when compared with control. Results from all the drug treated groups showed good sperm morphology, increased sperm count and motility. There was no DNA damage and showed normal chromosomal integrity even in 250 mg/kg dose. When compared with control all the three extract treated groups showed increased ROS scavenging activity. However, group II (200 mg/kg) showed significant changes in all the parameters. From the present study it was confirmed that the M. pruriens has potential to improve the sperm qualitatively and quantitatively through scavenging the excess ROS with any adverse side effects. These observations suggest that ethanolic seed extract of M. pruriens may serve as anti-oxidant that can exploit to treat the oxidative stress mediated male factor infertility.