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Protective Effect of [6]-Gingerol on the Ethanol-induced Teratogenesis of Cultured Mouse Embryos
연정민,남상윤,백인정,Se-Ra Lee,Mi-Ra Kim,홍진태,Hwanyul Yong,이범준,윤영원 대한약학회 2012 Archives of Pharmacal Research Vol.35 No.1
Excessive ethanol consumption during pregnancy causes fetal alcohol syndrome. We investigated the effect of [6]-gingerol on ethanol-induced embryotoxicity using a whole embryo culture system. The morphological changes of embryos and the gene expression patterns of the antioxidant enzymes cytosolic glutathione peroxidase (cGPx), cytoplasmic Cu/Zn superoxide dismutase (SOD1), and Mn-SOD (SOD2), and SOD activity were examined in the cultured mouse embryos exposed to ethanol (5 μL/3 mL) and/or [6]-gingerol (1×10−8 or 1×10−7 μg/mL) for 2 days. In ethanol-exposed embryos, the standard morphological score of embryos was significantly decreased compared with those of the control (vehicle) group. However, cotreatment of embryos with [6]-gingerol and ethanol significantly improved all of the developmental parameters except crownrump length and head length, compared with those of the ethanol alone group. The mRNA expression levels of cGPx and SOD2, not SOD1, were decreased consistently, SOD activity were significantly decreased compared with the control group. However, the decreases in mRNA levels of antioxidant enzymes and SOD activity were significantly restored to the control levels by [6]-gingerol supplement. These results indicate that [6]-gingerol has a protective effect against ethanol-induced teratogenicity during mouse embryogenesis.
Genistein Prevents Ethanol-Induced Teratogenesis in Mouse Embryos
연정민,Chunmei Lin,정아영,이종걸,정기연,백인정,이범준,윤영원,남상윤 사단법인 한국동물생명공학회 2011 한국동물생명공학회지 Vol.26 No.2
Drinking of excessive ethanol during pregnancy induces a fetal alcohol syndrome. Genistein is one of naturally occurring isoflavones at relatively high levels in soybeans. In this study, we investigated the effects of genistein (1×10^-8 and 1×10^-7μg/ml) on the ethanol (1μl/ml)-induced teratogenesis of developing mouse embryos during the critical period (embryonic days 8.5~10.5) of organogenesis using a whole embryo culture system and then morphological scoring analysis. Ethanol-treated embryos exhibited a variety of developmental abnormalities. However, the total morphological scores for ethanol plus genistein groups were significantly higher than those of ethanol alone group (p<0.05). In particular, there were significant increases in the ethanol plus 1×10^-8μg/ml of genistein group on the scores for heart, optic system, branchial bar, mandibular process, and caudal neural tube and further in the ethanol plus 1×10^-7μg/ml of genistein group on the scores for heart, hind-, mid-, and forebrains, optic system, branchial bars, maxillary and mandibular processes, caudal neural tube, forelimb, hindlimb, and somites as compared with those of ethanol alone group (p<0.05). These results indicate that genistein has a preventive effect against ethanol-induced teratogenesis.
연정민,Chunmei Lin,정아영,이종걸,정기연,백인정,이범준,남상윤,윤영원 사단법인 한국동물생명공학회 2011 한국동물생명공학회지 Vol.26 No.2
Genistein is a product of naturally occurring isoflavones at relatively high levels in soybeans. The harmful effects of ethanol are attributed to the induction of biological processes which lead to an increase in the generation of reactive oxygen species in fetuses. In this study, we investigated the effects of genistein (1×10^-8 and 1×10^-7μg/ml) on gene expressions of the representative cellular antioxidative enzymes in ethanol (1μl/ml)-treated mouse fetuses during the critical period (embryonic days 8.5~10.5) of organogenesis using a semi-quantitative RT-PCR analysis. The mRNA levels of cytosolic glutathione peroxidase (GPx), phospholipid hydroperoxide GPx, cytosolic Cu,Zn-superoxide dismutase (SOD), and mitochondrial SOD were significantly decreased in ethanol-treated fetuses. However, the mRNA levels of ethanol plus genistein-treated fetuses were significantly higher than those of ethanol alone fetuses. These results indicate that genistein can up-regulate the expressions of GPx and SOD mRNAs reduced by the ethanol treatment in fetuses.
Laser Capture Microdissection으로 절제된 마우스의 특정 단계별 정세관에서 Glutathione Peroxidase 유전자의 발현 분석
연정민,임춘매,박정훈,홍민기,정아영,김미라,백인정,이범준,남상윤,윤영원 한국발생생물학회 2010 발생과 생식 Vol.14 No.2
The seminiferous epithelium, with its division into 12 spermatogenic stages in the mouse, is a very complex tissue. Glutathione peroxidase (GPx) is a representative antioxidant enzyme that is capable of reducing organic hydroperoxides to their corresponding hydroxyl compounds utilizing glutathione and is related to the mammalian spermatogenesis. In this study, a real-time PCR was performed in the stage-specific seminiferous tubules of mouse testes excised by a laser capture microdissection (LCM) in order to quantitate the expression levels of a series of GPx genes including cytosolic GPx (cGPx), gastrointestinal GPx (GI-GPx), plasma GPx (pGPx), and phospholipid hydroperoxide GPx (PHGPx). Frozen sections (10 ㎛) were obtained from normal adult mouse testes. LCM was used to capture all the cells that were grouped into stages Ⅰ-Ⅴ, Ⅶ-Ⅷ, and Ⅸ-Ⅺ in cross-sections of seminiferous tubules. The expression level of PHGPx mRNA was remarkably higher than those of other GPx mRNAs in mouse testes. During spermatogenesis, the expressions of GI-GPx, pGPx, and PHGPx mRNAs were highest on stages Ⅶ-Ⅷ, began to decrease after stage Ⅺ, and showed a lowest level on stage Ⅰ-Ⅴ. However, the expressions of cGPx mRNA were highest on stages Ⅶ-Ⅷ, and showed a lowest level on stage Ⅺ-Ⅺ. These findings indicate that GPx genes are expressed differentially on mouse spermatogenesis and also LCM can be an useful tool in cellular quantitative analysis of testes.
연정민,Chunmei Lin,이윤복,이범준,윤영원,남상윤 사단법인 한국동물생명공학회 2012 한국동물생명공학회지 Vol.27 No.2
Linuron is a pesticide with a weak anti-androgenic property, which impacts male reproductive organs. In this study,to clarify whether linuron affects the cellular antioxidant system of ventral prostate, gene expression patterns of the representative antioxidant enzymes such as glutathione peroxidase (GPx), selenoprotein P (SePP), and superoxide dismutase (SOD) were investigated in the rat ventral prostates exposed to linuron using real-time RT-PCR analyses. Sprague-Dawley rats castrated at 6 weeks old were treated with linuron (25, 50, or 100 mg/kg per oral) daily for 10 days after testosterone propionate administration (0.4 mg/kg) subcutaneously. As compared to normal control animals,mRNA levels of phospholipid hydroperoxide GPx (PHGPx), SePP, and Mn SOD significantly increased in the prostates exposed to linuron (25, 50, and 100 mg/kg). However, cytosolic GPx (100 mg/kg) and Cu/Zn SOD (25, 50,and 100 mg/kg) mRNA levels significantly decreased in the ventral prostates. These results indicate that linuron upregulates the expressions of PHGPx, SePP, and Mn SOD mRNAs, but down-regulates the expressions of cytosolic GPx and Cu/Zn SOD in rat prostates, suggesting that linuron may have dual effects in the cellular antioxidant system of prostate.