• The Korean Journal of Physiology and Pharmacology
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• 제17권4호
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• pp.259-265
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• 2013

The anti-apoptotic effect of (-)-epigallocatechin-3-gallate (EGCG) during unilateral testicular torsion and detorsion (TT/D) was established in our previous study. In mice, the smallest inhibitor of apoptosis, survivin, is alternatively spliced into three variants, each suggested to have a unique function. Here, we assessed how EGCG exerts its protective effect through the expression of the different survivin splice variants and determined its effect on the morphology of the seminiferous tubules during TT/D. Three mouse groups were used: sham, TT/D+vehicle and TT/D treated with EGCG. The expression of the survivin variants (140 and 40) and other apoptosis genes (p53, Bax and Bcl-2) was measured with semi-quantitative RT-PCR. Histological analysis was performed to assess DNA fragmentation, damage to spermatogenesis and morphometric changes in the seminiferous tubules. In the TT/D+vehicle group, survivin 140 expression was markedly decreased, whereas survivin 40 expression was not significantly different. In parallel, there was an increase in the mRNA level of p53 and the Bax to Bcl-2 ratio in support of apoptosis induction. Histological analyses revealed increased DNA fragmentation and increased damage to spermatogenesis associated with decreased seminiferous tubular diameter and decreased germinal epithelial cell thickness in the TT/D+vehicle group. These changes were reversed to almost sham levels upon EGCG treatment. Our data indicate that EGCG protects the testis from TT/D-induced damage by protecting the morphology of the seminiferous tubules and modulating survivin 140 expression.

• Journal of Ginseng Research
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• 제43권1호
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• pp.125-134
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• 2019

Background: Excessive stress causes varied physiological and psychological disorders including male reproductive problems. Here, we attempted to investigate the protective effects of Korean Red Ginseng (Panax ginseng Meyer; KRG) against sub-acute immobilization stress-induced testicular damage in experimental rats. Methods: Male rats (age, 4 wk; weight, 60-70 g) were divided into four groups (n = 8 in each group): normal control group, immobilization control group, immobilization group treated with 100 mg/kg of KRG daily, and immobilization group treated with 200 mg/kg of KRG daily. Normal control and immobilization control groups received vehicle only. KRG (100 mg/kg and 200 mg/kg) was mixed in the standard diet powder and fed daily for 6 mo. Parameters such as organ weight, blood chemistry, sperm kinematic values, and expression levels of testicular-related molecules were measured using commercially available kits, Western blotting, and reverse transcription polymerase chain reaction. Results: Data revealed that KRG restored the altered testis and epididymis weight in immobilization stress-induced rats significantly (p < 0.05). Further, KRG ameliorated the altered blood chemistry and sperm kinematic values when compared with the immobilization control group and attenuated the altered expression levels of spermatogenesis-related proteins (nectin-2, cAMP responsive element binding protein 1, and inhibin-${\alpha}$), sex hormone receptors (androgen receptor, luteinizing hormone receptor, and follicle-stimulating hormone receptor), and antioxidant-related enzymes (glutathione S-transferase m5, peroxiredoxin-4, and glutathione peroxidase 4) significantly in the testes of immobilization stress-induced rats. Conclusion: KRG protected immobilization stress-induced testicular damage and fertility factors in rats, thereby indicating its potential in the treatment of stress-related male sterility.

• Clinical and Experimental Reproductive Medicine
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• 제50권3호
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• pp.160-169
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• 2023

Objective: Cryptorchidism is one of the main causes of infertility and can result in testicular cancer. This study aimed to present quantitative data on the damage caused by cryptorchidism using stereological analysis. Methods: Thirty newborn rats were randomly divided into control and experimental groups. The experimental group underwent surgery to induce unilateral cryptorchidism in the left testis, whereas the control group underwent a sham surgical procedure 18 days after birth. The testes were removed at designated time points (40, 63, and 90 days after birth) for stereological evaluation and sperm analysis. Total testicular volume, interstitial tissue volume, seminiferous tubule volume and length, and seminiferous epithelium volume and surface area were measured. Other parameters, such as sperm count, sperm morphology, and sperm tail length, were also examined. Results: Statistically significant differences (p<0.05) were observed between the experimental and the control groups at different ages regarding the volumes of various parameters, including the surface area of the germinal layer, the length of the seminiferous tubules, sperm count, and sperm morphology. However, no significant differences were observed in the epithelial volume and the sperm tail length of the groups. Conclusion: Given the substantial effect of cryptorchidism on different testicular parameters, as well as the irreversible damage it causes in the testes, it is important to take this abnormality seriously to prevent these consequences.

• 대한수의학회지
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• 제37권2호
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• pp.425-438
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• 1997

To assess the testicular toxicity induced by DA-125, a new anthracycline anticancer agent, the test substance was intraveneously administered to male beagle dogs at dose levels of 0, 0.0023, 0.0375, 0.15, and 0.6 mg/kg/day, 6 days a week for 26 weeks. At 0.6 mg/kg/day, 1 out of 3 dogs had died on day 42 of treatment and the other dogs were sacrificed on days 46 and 122 of treatment due to the increasingly severe clinical condition. Clinical signs considered to be related to treatment were included anorexia, vomiting, salivation, decreased activity, mucous and/or dark faeces, diarrhea, and swelling, abscess and/or ulceration of injection sites. Suppression in body weight gain, reduction in food intake, decreases in testicular weight and size, and hemorrhage of epididymis were also observed in male dogs. Microscopically, severe degenerative changes such as atrophy of seminiferous tubules, loss of germ cells, degeneration of germ cells, vacuolization of Sertoli cells, and hyperplasia of Leydig cells were observed in all dogs. Azoospermia in epididymal tubules, atrophy of epithelia in the cauda epididymis, and prostate atrophy were also found. At 0.15 mg/kg/day, anorexia, vomiting, salivation, diarrhea, and swelling of injection sites were observed. In addition, suppression in body weight gain and decreases in testicular weight and size were found in male dogs. Atrophy of seminiferous tubules, decrease of germ cells, degeneration, exfoliation and retention of germ cells, vacuolization of Sertoli cells, and hyperplasia of Leydig cells were observed by histopathological examination. Azoospermia in epididymal tubules and prostate atrophy were also found. At 0.0375 mg/kg/day, there were no clinical signs considered to be indicative of a reaction to treatment, but testicular size was significantly reduced. Microscopically, decreases in the number of spermatogonia and epidydimal speramtozoa were found. There were no evidences of general or testicular toxicity at 0.0023 mg/kg/day. These results indicate that DA-125 produces significant and persistent damage to the spermatogenic compartments of the testes in male beagle dogs.

• Clinical and Experimental Reproductive Medicine
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• 제35권2호
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• pp.131-141
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• 2008

목 적: 무정자증 불임부부에서 신선 (fresh) 고환정자 (testicular spermatozoa)와 냉동보존-융해(cryopreserved-thawed) 고환정자를 사용한 난자세포질내 정자주입술 (intracytoplasmic sperm injection, ICSI)의 결과를 비교하고자 하였다. 연구방법: 신선 고환정자 및 냉동보존-융해 고환정자를 사용하여 ICSI 시술을 시행하기로 계획된 총 109주기 (66명)를 대상으로 하였고 신선 고환정자를 사용하기로 계획한 군 (신선 고환정자군, fresh group)에는 92주기 (61명)이 포함되었고 냉동보존-융해 고환정자를 사용하기로 계획한 군 (냉동보존-융해 고환정자군, cryopreserved-thawed group)에는 17주기 (13명)가 포함되었다. 양 군간에 수정률, 착상률, 임신률, 유산률 등 ICSI 시술의 결과들을 비교하였고 통계학적 분석은 Mann-Whitney U 검정 및 Fisher의 정확한 검정을 적절하게 사용하였다. 결 과: 신선 고환정자를 사용하여 ICSI 시술을 시행하기로 계획된 총 92주기 중 9주기에서 고환정자를 추출할 수 없어 시술 주기가 취소되었다. 냉동보존-융해 고환정자군과 비교하여 신선 고환정자군에서 수정률이 높은 경향을 보였고 ($58.0{\pm}27.8%$ vs. $45.9{\pm}25.0%$, p=0.076) 양질의 배아 수는 통계적으로 유의하게 높았다 ($0.9{\pm}1.2$ vs. $0.2{\pm}0.5$, p=0.002). 그러나 임상적 임신율, 착상률, 유산율은 양 군간에 통계적으로 유의한 차이가 없었다. 결 론: ICSI 시술을 위하여 냉동보존-융해 고환정자를 사용하는 경우 수정률 및 배아의 질이 감소하지만 임신율, 착상률, 유산율에는 영향을 미치지 않는 것으로 사료된다. 또한, ICSI 시술이전에 고환정자를 확보하고 냉동보존-융해 고환정자를 사용한다면 난자채취 당일 정자를 확보하지 못하여 주기를 취소하는 경우나 여성배우자의 불필요한 과배란유도를 줄일 수 있으며 반복적인 고환정자추출술로 인한 고환기능의 손상을 줄일 수 있는 유용한 방법으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제50권1호
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• pp.19-25
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• 2023

Objective: Sleep deprivation (SD) is a common problem in today's stressful lifestyle and have physiological consequences, including reproductive dysfunction and infertility. As an antioxidant, olive oil may be effective in reducing testicular and spermatological damage by decreasing the production of free radicals. Methods: This study investigated the effects of olive oil on sperm quality and testicular structure using stereological methods to assess rats with SD. Results: When comparing SD group to grid floor+distilled water (GR) group, we found that the sperm count and motility, as well as the percentage of slow progressive sperm was significantly lower in SD group (p<0.05), but the percentage of immotile sperm was higher (p<0.01). However, no improvement was observed in sperm count or motility after concomitant treatment of SD group with olive oil. Stereological examinations revealed no significant change in the total volumes of the seminiferous tubules, interstitial tissue, and germinal epithelium in the study groups. Conversely, the total number of testicular cell types was significantly lower in SD group than in GR group. Although the total number of Sertoli and Leydig cells was significantly higher in the S +olive oil group than in the untreated SD group, no significant difference in the total number of other testicular cell types was observed between the two groups. Conclusion: SD potentially induced structural changes in testis that affected sperm count and motility. However, olive oil only improved the total number of Sertoli and Leydig cells in the animals with SD and did not improve sperm count and motility.

• 대한임상검사과학회지
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• 제41권3호
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• pp.111-122
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• 2009

Cadmium (Cd) is known to exert gonadotoxic and spermiotoxic effects. The present study was performed to investigate the morphological effects and metallothionein (MT) expression by zinc pretreatment in the course of time of cadmium-induced testicular injury in rat. Fifty male Spraque-Dawley rats weighing 160~180 g were divided into two groups : saline-pretreated cadmium group and zinc-pretreated cadmium group. Rats of two groups received subcutaneous injection of saline and 100 mg/kg $ZnSO_4$ at 0, 2, 5 and 8 hrs intervals respectively. Cadmium chloride (4.5 mg/kg $CdCl_2$) was administrated intraperitoneally at 2 hrs after zinc injection and rats were killed 0, 12, 24, 48 and 72 hrs later. Testicular tissue damages, interstitial (Leydig) cells status and MT expression were determined using hematoxylin-eosin stained sections and a computerized image analysis system on sections immunostained with a mouse anti-metallothionein respectively. Zinc pretreatment was significantly reduced testicular damages in five pathological categories after cadmium administation. The number of surviving interstitial cells was significantly higher in the zinc-pretreated group than in the saline-preatreated group at 48 and 72 hrs after cadmium administration. Non-damaged testis showed the positivity of MT staining in spermatogenic cells, Sertoli cells and endothelium of blood vessel, but not in the Leydig cells. The positivity of MT staining in saline-pretreated group was significantly reduced at 24 hrs after cadmium administration, whereas zinc-pretreated group showed strong MT positive staining similar to the 0 hr by 42 hrs after cadmium administration. In damaged testis, MT positive staining was also observed in the Leydig cells of both groups. These results suggest that a major preventive effect of zinc against cadmium-induced testicular toxicity may be due to its ability to reduce the cytotoxicity of cadmium in spermatogenic cells and Leydig cells by inhibiting the susceptibility of the testis to cadmium but not MT production by cadmium.

• 대한임상검사과학회지
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• 제42권3호
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• pp.136-148
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• 2010

Cadmium (Cd) is known to exert gonadotoxic and spermiotoxic effects. The present study was performed to investigate the morphological effects and metallothionein (MT) expression by zinc pretreatment in the course of time of cadmium-induced testicular injury in rat. Fifty male Spraque-Dawley rats weighing 160-180 g were divided into two groups: saline-pretreated cadmium group and zinc-pretreated cadmium group. Rats of two groups received subcutaneous injection of saline and 100 mg/kg $ZnSO_4$ at 0, 2, 5 and 8 hrs intervals respectively. Cadmium chloride (4.5 mg/kg $CdCl_2$) was administrated intraperitoneally at 2 hr after zinc injection and rats were killed 0, 12, 24, 48 and 72 hrs later. Testicular tissue damages, Interstitial (Leydig) cells status and MT expression were determined using hematoxylin-eosin stained sections and a computerized image analysis system on sections immunostained with a mouse anti-metallothionein respectively. Zinc pretreatment was significantly reduced testicular damages in five pathological categories after cadmium administation. The number of surviving interstitial cells was significantly higher in the zinc-pretreated group than in the saline-preatreated group at 48 and 72 hrs after cadmium administration. Non-damaged testis showed the positivity of MT staining in spermatogenic cells, Sertoli cells and endothelium of blood vessel, but not in the Leydig cells. The potitivity of MT staining in saline-pretreated group was significantly reduced at 24 hrs after cadmium administration, whereas zinc-pretreated group showed strong MT positive staining similar to the 0 hr by 42 hrs after cadmium administration. In damaged testis, MT positive staining was also observed in the Leydig cells of both groups. These results suggest a major preventive effect of zinc against cadmium-induced testiculat toxicity may be due to its ability to reduce the cytotoxicity of cadmium in spermatogenic cells and Leydig cells by inhibiting the susceptibility of the testis to cadmium but not MT production by cadmium.

• Journal of Ginseng Research
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• 제34권4호
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• pp.327-335
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• 2010

We reported previously that the administration of Korean red ginseng water extract (KRG-WE) protected the guinea pig testis against damage induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (a potent endocrine disruptor). We also found that crude saponin from ginseng was the active ingredient responsible for this protection. Here, we examined the biological role of KRG-WE in an animal model of age-induced dysfunction of spermatogenesis. Twenty-four male Sprague-Dawley (six 2-month-old and eighteen 12-month-old) rats were used. The young and old control groups received only vehicle. The ginseng saponin (GS)- and KRG-WE-treated groups received GS (40 mg/kg body weight/day) and KRG-WE (200 mg/kg body weight/day), respectively, for 4 months. The number of cells, Sertoli cell index, Johnsen's score, and sex hormone levels decreased significantly with age. However, the administration of KRG-WE and GS markedly improved the number of germ cells, seminiferous tubular size, and Johnsen's score in the old rats. Ginseng produced a distinct testicular histological improvement in old rats. KRG-WE and GS elevated testosterone levels, while attenuating the aberrant increase in follicle stimulating hormone and luteinizing hormone levels. Sperm kinematics evaluated by a computer-assisted sperm analyzer demonstrated improvement in the percentage of motile sperm, progressive sperm motility, and curvilinear velocity associated with sperm quality, supporting the beneficial role of red ginseng in senile spermatogenesis. Overall, the total water extract had a more potent effect than the corresponding saponin fraction. In conclusion, Korean red ginseng rejuvenated age-induced testicular dysfunction. Additionally, the total water extract was more potent than the corresponding saponin fraction.

• 한국동물생명공학회지
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• 제38권4호
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• pp.189-198
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• 2023

Background: Brassica oleracea var. italica (broccoli), a rich source of antioxidants, can prevent various diseases and improve human health. In this study, we investigated the antioxidative effects of broccoli sprout extract on oxidative stress induced by lipopolysaccharide and cisplatin in cell and organ tissue models. Methods: Antioxidative effect of BSE was evaluated using DPPH and ABTS in RAW 364.7 cells, and effects of BSE on testes were investigated using Cisplatin-induced testicular damage model with an in vitro organ culture system. Results: The DPPH assay showed that the antioxidant activity of the alcoholic broccoli sprout extract was higher than that of the water extract. Additionally, the expression levels of antioxidation-related genes, Nrf2, Gsr, HO-1, and catalase, were significantly increased in broccoli sprout extract-treated RAW 264.7 cells, and the extract suppressed lipopolysaccharide-induced mitochondrial dysfunction. Based on the results in the RAW 264.7 cell culture, the antioxidative effects of the extracts were investigated in a mouse testis fragment culture. The expression of Nrf2, HO-1, and Ddx4 was clearly decreased in cisplatin-treated mouse testis fragments and not in both broccoli sprout extract- and cisplatin-treated mouse testis fragments. In addition, the oxidative marker O-HdG was strongly detected in cisplatin-treated mouse testis fragments, and these signals were reduced by broccoli sprout extract treatment. Conclusions: The results of this study show that broccoli sprout extracts could serve as potential nutraceutical agents as they possess antioxidant effects in the testes.