3-MCPD의 생식ㆍ발생독성에 관한 연구

곽승준(Seung Jun Kwack),김순선(Soon Sun Kim),최요우(Yo Woo Choi),이규식(Gyu Seek Rhee),손경희(Kyung Hee Sohn),이이다(Rhee Da Lee),채수영(Soo Young Chae),정용현(Yong-Hyun Chung),유일재(Il Je Yu),박귀례(Kui Lea Park) 한국독성학회 2004 Toxicological Research Vol.20 No.1

3-Monochloro-1,2-propanediol(3-MCPD) is a toxic compound, often present in different foods containing acid hydrolyzed(AH) protein, like seasonings and savory food products. The purpose of the present study was to investigate the effects of 3-MCPD on male fertility, sperm and testosterone secretion. In vivo male fertility test was performed for observing the adverse effects of 3-MCPD on the function of male reproductive system and pregnancy outcome. 0.01, 0.05, 0.25, 1 and 5 mg/kg b.w. of 3-MCPD was given daily by gavage to groups of 15 adult male SD rats for 4 weeks. At the end of pre-treatment period, males were mated overnight with normal females. Following morning, males demonstrating successful induction of pregnancy were sacrificed on that day to assess<br/> sperm parameters and histopathology of reproductive organs. The resulting pregnant females were sacrificed on day 20 of gestation to evaluate pregnancy outcome. As a result, four-week paternal<br/> administration with 3-MCPD resulted in adverse effects on male fertility and pregnancy outcome without remarkable histopathological changes in testes and epididymides; sperm motility, copulation index and fertility index were markedly decreased in the treated group and numbers of live fetuses showed<br/> steep dose-response curves. Also, spermatogenesis was investigated in this experiment. However, no effect was observed on production of sperm in testes treated with 3-MCPD for 4 weeks. Hormone assay was performed for observing the effects of 3-MCPD on testosterone and luteinizing hormone (LH) in blood and testes of male SD rats and cultured primary Leydig cell. In result, significant changes of related hormones did not observed by treatment of 3-MCPD. These results indicated that paternal treatment with 3-MCPD induced spermatotoxic effect, which caused an antifertility on male.

3-MCPD의 생식˙발생독성에 관한 연구

곽승준(Seung Jun Kwack),김순선(Soon Sun Kim),최요우(Yo Woo Choi),이규식(Gyu Seek Rhee),손경희(Kyung Hee Sohn),이이다(Rhee Da Lee),채수영(Soo Young Chae),정용현(Yong-Hyun Chung1),유일재(Il Je Yu1),박귀례(Kui Lea Park) 한국독성학회 2004 Toxicological Research Vol.20 No.2

3-Monochloro-1,2-propanediol(3-MCPD) is a toxic compound, often present in different foods containing acid hydrolyzed(AH) protein, like seasonings and savory food products. The purpose of the present study was to investigate the effects of 3-MCPD on male fertility, sperm and testosterone secretion. In vivo male fertility test was performed for observing the adverse effects of 3-MCPD on the function of male reproductive system and pregnancy outcome. 0.01, 0.05, 0.25, 1 and 5 mg/kg b.w. of 3-MCPD was given daily by gavage to groups of 15 adult male SD rats for 4 weeks. At the end of pre-treatment period, males were mated overnight with normal females. Following morning, males demonstrating successful induction of pregnancy were sacrificed on that day to assess sperm parameters and histopathology of reproductive organs. The resulting pregnant females were sacrificed on day 20 of gestation to evaluate pregnancy outcome. As a result, four-week paternal administration with 3-MCPD resulted in adverse effects on male fertility and pregnancy outcome without remarkable histopathological changes in testes and epididymides; sperm motility, copulation index and fertility index were markedly decreased in the treated group and numbers of live fetuses showed steep dose-response curves. Also, spermatogenesis was investigated in this experiment. However, no<br/> effect was observed on production of sperm in testes treated with 3-MCPD for 4 weeks. Hormone assay was performed for observing the effects of 3-MCPD on testosterone and luteinizing hormone (LH) in blood and testes of male SD rats and cultured primary Leydig cell. In result, significant changes of related hormones did not observed by treatment of 3-MCPD. These results indicated that paternal treatment with 3-MCPD induced spermatotoxic effect, which caused an antifertility on male.

Thimerosal의 발생독성에 관한 연구

곽승준(Seung Jun Kwack),이규식(Gyu Seek Rhee),김순선(Soon Sun Kim),손경희(Kyung Hee Sohn),김소희(So Hee Kim),채수영(Soo Young Chae),최요우(Yo Woo Choi),원용혁(Yong Hyuck Won),박귀례(Kui Lea Park) 한국독성학회 2003 Toxicological Research Vol.19 No.4

The purpose of our study was to evaluate the toxicity of the thimerosal in embryos and neonates. Thimerosal (also known as mercurothiolate) is a mercury-containing compound used in<br/> trace amounts to prevent bacteria and other organisms from contaminating vaccines, especially in opened multi-dose vials. The toxicity of mercury is well known and those most at risk occurrs in<br/> unborn babies and newborn babies. Test methods included in vitro whole embryo culture (WEC) system and in vivo test of neonatal toxicity in Wistar rats. Ethylmercury and methylmercury were used as positive controls for the evaluating of toxic effects of mercury. In WEC assay, treated concentrations of thimerosal, ethylmercury and methylmercury were up to 0.01, 0.025, 0.05, 0.1, 0.25, 0.5, 1, 2.5 and 5 mg/ml, respectively. All compounds didn’t show any morphological abnormalities, but showed retardation<br/> of growth and development in dose dependent manner (>0.5㎍/ml). These data indicated that thimerosal showed developmental toxicity in vitro. In vivo neonatal toxicity, Wistar rats were administered<br/> subcutaneously with thimerosal, ethylmercury, or methylmercury (5, 25, 50, 250, and 500 mg/kg) during from postnatal day (PND) 4 to 25. Significant effects of these compounds on relative organ<br/> weights and organ morphology were not observed in this experiment. However, accumulation of mercury was detected in the kidney and testis when treated with thimerosal, ethylmercury, or methylmercury. These results suggest that thimerosal may be a harmful compound to embryo and neonate, but used concentration of thimerosal in these experiments is much higher than that of clinical application. Further investigation is needed on the safety of vaccine components, i.e. a thimerosal using in vitro and in vivo tests in the future.

선천성 사지기형 발생 기전 연구(Ⅲ) : Hox유전자 발현에 관한 연구 Studies on the Expression of Hox gene

손경희,권오란,김순선,김소희,곽승준,김병호,최요우,김태성,김형식,신재호,문현주,김명희,한순영,하광원,박귀례 식품의약품안전청 2000 식품의약품안전청 연보 Vol.4 No.-

Homeobox-containir을 (Hox) 운전자는 발생단케, 위치 및 조직 특이적인 발현에 따라 배자의 발생 과정동안 형태형성에 관여하는 것으로 알려져 있다. 딘ok 유진자들(또oxa-1, Hoxa-2, Hoxb-6 Hoxa-7, Hoxc-8) 중 특히 Hoxa-7 유전자는 ectoderm에서 유래하는 neural tube 및 neoral crest ceIBs과 mesoderm에사 유래하는 prevertebratae에서 전 ·후축 형성 시에 발현하는 것으로 잘 알려진 유전자이다. 그러나 Hol(a-? 유전자가 배자의 사지 및 꼬리 발생에 직접적으로 관여하는지에 대해서는 아직까지 잘 알려지지 않았다. 따라서 본 연구에서는 최기형성 물질로 알려진 all-trans retinoic acid (RA)하 cyclophosphandde (CP)를 임신 9.5일의 마우스에 투여한 후 임신 10.5, 11.5, 12.5, 13.5 및 14.5꿀에 배자를 적출 f':4였을 때 나타나는 형태학적 이상과 Hoxa·-7 유전자 발현의 위치와의 상관성을 비교하기 위하여 whoBe mount in situ hybridization(WISH)을 실시하였다. 그 결과, 템oxa-7의 antisense RNA probe를 사용하였을 경우, 용매대조군과 비교하여 RA 투여 시 나타난 형태학적 이상인 꼬리단락의 위fl됐 「loxa-7이 강하게 발현되었고, CP 투여 시에는 용매대조군과 비교하여 배자꼭 크기가 현저히 적어서, 전체적인 발달이 저해된 것으로 나타났으며, 특히 발달이 저해된 foreBimb과 hindlimb의 위치에 Hoxa-7이 강하게 발현되었다 한편, antisense RNA probe의 음성대조군인 Hoxa-7의 sense RNA 1)robe로 WISH를 실시한 결과 antisense probe를 사용한 결과와는 달리 색깔 반응이 현저히 약하게 나타났다. 파라서 본 연구결과, 최기형성 물질에 노출시 나타나는 마우스 배자의 형태학적이상은 Hoxa-7 유전자의 발현 증가와 상관성이 있는 것으로 추정된다. Homeobox-containing (HOX) genes are known to be involved in pattem formation during development by being expressed in a stage-, position-, and tissue-specific manner, Aoong Hox genes (Hoxa-1, Hoxa-2, Hoxa7, Hoxb-6, Hoxc-8), Hoxa-? is one of the well 3tnown genes, whicll expressed In the ectoderm-defved neural tube and neural crest cells,and mesoderm-derived frevertebrae along the anterior-posterior axis. However, it is not clearwhether Hoxa-7 gene if; direcfBy involved in mouse embryonic timb and tail development.0order to establish the correlation between Hoxa-7 gene expression and the effect of teratagensduring mouse embryogenesis, we have analyfed Hoxa-7 gene expression in the developing limbbuds and tails of ICR mouse embryes (gestation day 10,5, 11,5, 12.5, 12.5, 14.5) after maternaladministration of teratogens, atl-trans retineic actd (RA) and cycBophosphahEde (CP), ungestation day 9.5 using whole mount in situ hybridization (WISH). We f4nd that RA causesshortening and crookedness of tail and CP causes develoPmental retardation of embryo,especia31y Ihfb buds. Rerutts from IfnSH show that expression of Hoxa-7 is increased in thetail of embryos which were treated with RA. Whereas Hoxa-7 gene expression is increased inthe limb buds of embryos, wich were treated with CP compared to vehicle control group.Taken together, these data suggest that the pattern of Hoxa-7 erpression correlates with theabnormality caused by ter;3togens during the mouse embryogenesis.

아리스톨로크산 함유 생약제에 대한 안전성평가연구 : 생식·발생독성시험연구 Studies on the reproductive and developmental toxicity

손경희,이규식,김순선,김소희,곽승준,채수영,박철훈,김병호,최요우,이종필,제금련,강진석,장동덕,길광섭,최광식,박귀례 식품의약품안전청 2001 식품의약품안전청 연보 Vol.5 No.-

아리스톨로크산(ArA)은 Jlristolochia 속 식물에 많이 항유되어있는 성분으로 이 성분이 들어있는 약을 장기복용한 사람들에게서 신장암 등 비뇨기계 암의 발병를이 놀은 것으로 알려졌다. 본 연군에서는 아리스톨로크산 함유 생약제인 마두령의 안전성에 대한 촤학쩍 근거를 마련하기 위하여 마두령 단방과 마두령 복방에 대한 생식 ·살생독성시험을 실시하였다. 마두령 단방 건조추출물(0.j, 2, 8mg/kg; ArA 릉도) 및 복방 건조춘출물(0.05, 0.Smg/ifg; ArA 농도)을 젓드에 임신 전 14일부터 기간 항성기인 임신17일까지 경구 투여한 후 잉신 ?0일에 제왕절개하여 모체와 태자에 대한 검사를 실시하고 이를 아리스톨로크산(2, 8, 16mg/k끄 투여 실험과 비교하였다. 마두령 단방 8mg트 투여군의 체중증가량과 사접춰량이 대조군에 비해 감소하였으며, 아리스톨로르산은 8m을J'k핑 및 15m료토 투여시 임신 전 ·후의 체중증가량 및 사료섭취량이 모두 대조군에 비해 감소하옅다. 또한. 마두령 단방의 경우 8mgg으 투여시 잉신이 저해되는 것_근로 나타났으나. 아리스톨로크산은 임신율에 영향을 미치지 않았고 다민 16mgAg 투여군에서 태자 체중이 유의성있게 강소하였다. 마두령 단방 8m9,/kg 투여군곽 아리스톨크산 8mg/kg 및 16mgg 투여군에서 간장의 절대무게 강소, 진장의 장대무게 증가, 비장의 절대무게감소 등이 간찰되었으며, 조직병리학적검사에서는 마두령 단방 8m9./k9 투여관과 아리스톨로크 16mgg 투여군에서 신장의 손상이 나타났다. 전배자 배양 실험에서는, 마두령 단방은 Ir끄ml 이상에서 태자발달이 현저히 저해되었고, 마두령 복방의 겅우 0.Ofrg/ml 이상에서, 아리스톨로크산끈 경우 300r9/r리 이상의 농도에서 배자발달이 저해되는 것으로 나타났다. 따라4i 본 연구결과로 띠루어 마두령은 임상용량인 0.03mg (.f.rA)/kg/da)-에서는 생식 · 발생득성을 일으키지 않는 것으로 사료된다. Rapidly progressive interstitial renal fibrosis has recently been reported in young women who have been on a slimming regimen including chinese herbs. Aristolochic acid, suspected as the causal factor of this renal disease, is a well known carcinogen. It has been known that Madouling (Aristolochiae fructus) contains aristolochic acid. The objective of this study was to investigate the effects of Madouling, Madouling-tang, and aristolochic acid on reproductive and developmental toxicity. Female rats were administered orally with the extracts of Madouling (0.5, 2, 8mg/kg;ArA dose). madouling-tang (0.05, 0.5mg/kg), and aristolochic acid(2, 8, 16mg/kg) from 14 days before mating to day 17 of gestation. Madouling(8mg/kg) decreased fertility in the 8mg/kg group, but Madouling-tang and aristolochic acids did not. Significant decrease of mean fetal body weights were observed in the 16mg/kg group of aristolochic acids. External, visceral and skeletal malformation of fetuses were not observed with treatment. There were decrease in the absolute liver weights and the absolute spleen weight and increase in the relative kidney weights in the 8mg/kg group of Madouling and 8 and 16mg/kg groups of Aristolochic acid. Histopathological examination showed the discrete damage of kidney in the 8mg/kg group of Madouling and 16mg/kg groups of Aristolochic acid. In whole embryo culture, Madouling and Madouling-tang caused the retardation of growth and development of embryo in the dose of 1㎍/ml and 0.02㎍/kg, respectively while Aristolochic acids showed the similar effect in the dose of 300㎍/kg. These results indicate that Madouling, up to 0.05mg/kg (prescription dose to human) has no adverse effects on the fertility, reproduction and development of Sprague-Dawley rats.