• Toxicological Research
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• 제14권2호
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• pp.143-149
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• 1998

This study was carried out to evaluate the testicular toxicity of 2-bromopropane (2-BP), which recently caused occupational intoxication on the reproductive and hematopoietic system in Koreans, using light microscopy, immunohistochemistry and flow cytometry. 10 weeks old male Sprague-Dawley (SD) rats were treated with 0.5 g/㎏/day of 2-BP orally for 8 consecutive weeks. The testes of the rats were vascularly perfused with Karnovsky's solution or immersed in Bouin's solution, embedded in plastic and evaluated with light microscopy. And relative proportions of haploid, diploid, and tetra-ploid states of DNA ploidy in the testicular cell suspensions of the SD rats were examined by flow cytometry. 2-BP induced severe testicular atrophy, depletion and degeneration of spermatogonia, spermatocytes, and spermatids and mild hyperplasia of Leydig cells without significant morphological changes. The Leydig cell hyperplasia was confirmed by immunohistochemistry using proliferating cell nuclear antigen (PCNA). The immunopositive cells against PCNA were observed in the nuclei oj some interstitial cells. Relative proportions of haploid states of DNA ploidy decreased in the atrophic testicular cell suspensions comparing with those of the control. In conclusion, 2-BP induced testicular atrophy with Leydig cell hyperplasia as examined by histopathology, immunohistochemistry and DNA flow cytometry.

• Toxicological Research
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• 제16권2호
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• pp.163-172
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• 2000

Recently, there is a worldwide concern that a great number of man-made chemicals have a hormone-like action both in humans and in animals. DECD is developing screening programs using validated test systems to determine whether certain substances may have an effect in humans. In the present study. the establishment oj repeated-dose toxicity test method was tried. Flutamide. an anti-androgenic agent. was administered by gavage to Sprague-Dawley rats for 28 days at dose levels of 0. 0.5. 3 and 18 mg/kg body weight (10-15 rats/sex/group) to examine the effects on general findings. especially reproductive and endocrine parameters. Clinical signs. body weights, food consumption, and sexual cycle were checked and measured. For the gross and microscopic examinations. 10 rats/sex/group were sacrificed at the end of dosing period and the remaining animals of control and high dose groups (5 each) were sacrificed after 14 days recovery. Examinations for hematology and clinical chemistry were carried out at necropsy. There were no treatment-related changes in clinical signs. body weights, food consumption. gross necropsy. hematology and clinical chemistry at all doses of both sexes. The period and regularity of sexual cycle were not adversely affected at all doses by the test agent. At 18 mg/kg. both decreased weights of prostate, seminal vesicle and epididymis in males and increased weights of spleen and thymus in females were observed. In addition, decreased number of spermatids and sperms. increased serum testosterone concentration and increased incidence (100%) of interstitial cell hyperplasia were seen in males. At 18 mg/kg of the recovery group. decreased prostate weight. reduced sperm count and increased incidence (20%) of interstitial cell hyperplasia in males and increased thymus weight in females were observed. At 3 mg/kg. reduced sperm count was found. There were no adverse effects on parameters examined at 0.5 mg/kg of both sexes. The results suggested that the potential target organs of flutamide may be accessory sexual glands including testes for males and spleen and thymus for females. Taken together. this test method was found to be a useful screening test system for endocrine disrupting chemicals.

• 생명과학회지
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• 제28권10호
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• pp.1147-1155
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• 2018

신령버섯균사체 액체배양 추출물(Agaricus blazei mycelial liquid culture extract, ABMLCE)은 TM-3 mouse Leydig 고환세포에서 testosterone (TS)의 생성을 촉진하였다. 이번 연구에서는 eritadenine (EA, 15.3 mg/100 g)을 함유한 ABMLCE가 RWPE-1 전립선세포에서 전립선비대증(Benign prostatic hyperplasia, BPH) 관련 핵심 효소인 $5{\alpha}-reductase$ 2 ($5{\alpha}-R2$) 활성과 dihydrotestosterone (DHT) 함량을 감소시켰음을 보고한다. RWPE-1 전립선 세포는 24-well plate에서 ABMLCE (0~50 ppm), EA (0~10 ppm,), finasteride (FS $10{\mu}M$: positive Control)를 함유한 Keratinocyte serum-free medium (K-SFM)에서 24시간 배양하였다. 시료처리 세포배양액에 대해 $5{\alpha}-R2$, superoxide dismutase (SOD), catalase (CAT), cyclooxygenase-2 (COX-2) 효소활성, TS, DHT, tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$)와 $interleukin-1{\beta}$ ($IL-1{\beta}$)를 assay kit를 사용하여 분석하였다. $5{\alpha}-R2$ 활성과 DHT 함량은 ABMLCE의 농도 의존적으로 유의성(p<0.05) 있게 감소되었고, SOD와 CAT 효소활성은 유의성(p<0.05) 있게 증가되었다. 반면에 COX-2 활성과 $TNF-{\alpha}$ 및 $IL-1{\beta}$의 함량은 감소되었다(p<0.05). 마찬가지로 EA의 효과도 ABMLCE 효과와 유사하였다. ABMLCE 50 ppm과 EA 10 ppm의 $5{\alpha}-R2$ 및 DHT 감소효과는 FS $10{\mu}M$ 효과와 유사하였다. 이들 결과는 ABMLCE와 EA는 항염증효과와 항산화효과를 통하여 BPH 유발 핵심 인자인 $5{\alpha}-R2$ 효소활성과 DHT 함량을 감소시켰다. 또한 이 결과는 EA를 함유하는 ABMLCE는 인체 BPH 개선 소재로 활용할 수 있음을 의미한다.

• Environmental Analysis Health and Toxicology
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• 제15권4호
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• pp.123-130
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• 2000

택트 스위치 제조공정 침지액의 주성분인 2-bromopropane독성에 대한 연구로 최단기의 폭로로 농도를 달리하여 3주간 반복 투여 시험을 시행하여, 흰쥐의 혈액 및 세정관의 변화를 관찰하기 위해 투여기간 동안의 체중의 변화, 고환, 간, 신장 등의 장기무게, 혈액화학과 혈액학적 분석 및 고환의 병리조직의 변화 등을 관찰하여 2-bromopropane의 급성투여 조건에서의 중독현상을 비교.분석하였다. 농도를 달리하여 투여에 따른 체중변동은 통계적으로 유의한(P<0.05)체중의 감소를 나타내었다. 1.000 mg/kg 투여군에서 백혈구수, 적혈구수, 혈구용적과 혈색소 농도에서 유의한 변화(p<0.05)를 보였다. 조직병리학적 소견으로 정 소에서는 세정관의 정조세포와 정모세포의 괴사를 볼 수 있었고, 기저막의 비후, 세정관의 Sertoli세포는 광범위하게 세포질성 공포현상을 보여주고 있다. 또한 간질조직에서는 Leydig 세포의 증식을 볼 수 있었다. 2-bromopropane의 손상부위는 조혈과 생식계가 표적으로 생각되며, 고농도 투여가 저농도 투여에 비해 독성이 심하며 독성물질의 양-반응 관계를 보여주고 있다.

• 대한수의학회지
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• 제40권2호
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• pp.361-371
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• 2000

최근 2-bromopropane(2-BP)이 사람과 실험동물에서 정소독성을 유발한다고 보고된 바 있다. 그러나 수컷 생식기계에 있어서 2-BP의 지연효과에 대해서는 세부적으로 조사된 바가 없다. 본 연구는 Sprague-Dawley 랫드에서 2-BP의 정소독성과 정자발생의 회복을 조사하기 위하여 수행하였다. 5주령의 수컷 랫드에게 2-BP를 1,000mg/kg 용량으로 4주간 반복투여하였고, 투여시작후 1, 2, 3, 4 및 12주째에 부검하였다. 정소독성의 평가는 병리조직학적인 질적평가와 생식기관 중량, 정자두부수 및 재생지수 등의 양적평가로 수행하였다. 시험결과 2-BP를 투여한 랫드에서는 체중과 정소 및 정소상체 중량이 대조군에 비해 시간의존적인 방식으로 억제 또는 감소하였다. 병리조직검사에서는 투여 1주째에 stage I~IV에서 정조세포와 stage VII~IX에서 세사전기 및 세사기의 정모세포가 현저하게 소실되었다. 정조세포는 투여 2주째에 모든 stage에서 광범위하게 소실되었으며, 정자발생주기가 진행됨에 따라 2, 3 및 4주째에는 접합기 정모세포, 비후기 정모세포 및 원형 정자세포가 전구세포의 결손에 의해 점진적으로 소실되었다. 지지세포의 기능적 이상을 암시하는 지지세포의 공포화와 정자세포 저류는 상기한 모든 시기에서 관찰되었다. 8주 회복후인 12주째에는 대부분의 곡세정관이 심하게 위축되어 지지세포만 관찰되었으며, 간질조직에서는 간질세포의 과형성이 인정되었다. 또한 2-BP에 의해 유발된 정소의 손상이 비가역적임을 암시해주는 정자두부와 재생지수의 현저한 감소가 관찰되었다. 상기결과는 랫드의 2-BP를 1,000mg/kg의 용량으로 4주간 반복투여하면 정조세포의 결손에 의해 점진적으로 생식세포가 감소하고 이로 인하여 장기적인 정소위축이 유발된다는 것을 암시해준다.

• Clinical and Experimental Reproductive Medicine
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• 제29권4호
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• pp.303-310
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• 2002

Objective s: To estimate the frequency of Y chromosome microdeletions in the Korean population of infertile men and to evaluate the relationship between microdeletion on the Y chromosome and clinical phenotypes of infertile men with idiopathic azoospermia and oligozoospermia. Materials and Methods: Genomic DNA was extracted from blood samples collected from 330 infertile men attending the Infertility Clinic at Samsung Cheil Hospital, Korea. Six sequence tagged sites (STSs) spanning the azoospermia factor (AZF) regions of the Y chromosome were amplified by polymerase chain reactions (PCRs). Results: Microdeletions on Y chromosome were detected in 35 (10.6%) of the 330 infertile men. Most of the microdeletions (91.4%) involved AZFb or AZFc. The high incidence of microdeletions were found in AZFc region (57.1%), but the low in AZFa (8.6%) and AZFb (5.7%). Larger microdeletions involving two or three AZF regions were detected in 28.6% of cases. All patients (6 patients) with deletion of AZFa region showed no germ cell phenotypes, Sertoli cell only syndrome or Leydig cell hyperplasia in histopathologic examinations. Conclusion: Microdeletions on the Y chromosome, especially, at AZFc/DAZ regions may be the major cause of azoospermia and severe oligozoospermia. We suggest that idiopathic infertile men have genetic counselling and microdeletion analysis on the Y chromosome before IVF-ET and ART program.

• Journal of Genetic Medicine
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• 제2권1호
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• pp.11-15
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• 1998

This paper reports 3 cases with 46,XX sex reversed male. Three 46,XX hypogonadal subjects showed complete sex reversal and had normal phallus and azoospermia. We studied them under clinical, cytogenetic and molecular aspects to find out the origin of the sex reversal. Patients had markedly elevated serum follicle-stimulating hormone (FSH) and lutenizing hormone (LH) and decreased or normal range of serum testosterone. The testicular volumes were small (3-8ml). Testicular biopsy showed Leydig cell hyperplasia and atrophy of seminiferous tubules. We obtained the results of normal 46,XX, and the presence of Y chromosome mosaicism was ruled out through XY dual fluorescent in situ hybridization (FISH). By using polymerase chain reaction (PCR), we amplified short arm (SRY, PABY, ZFY and DYS14), centromere (DYZ3), and heterochromatin (DYZ1) region of the Y chromosome. PCR amplification of DNA from these patients showed the presence of the sex-determining region of the Y chromosome (SRY) but didn't show the centromere and heterochromatin region sequence. The SRY gene was detected in all the three patients. Amplification patterns of the other regions were different in these patients; one had four amplified loci (PABY+, SRY+, ZFY+, DYS14+), another had two loci (SRY+, ZFY+) and the other had two loci (PABY+, SRY+). We have found that each patient's translocation elements had different breakpoints at upstream and downstream of the SRY gene region. We conclude that the testicular development in 46,XX male patients were due to insertion or translocation of SRY gene into X chromosome or autosomes.