논문 상세보기
pp.45-51
Background: This study was performed to evaluate the protective effect of Saururus chinensis ethanol extract (SCE) against styrene toxicity in mouse spermatocyte cells [GC-2spd (ts) cell line].
Methods and Results: Cytotoxicity in mouse spermatocyte cells was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Generation of reactive oxygen species (ROS) was determined using 2’,7’-dichlorodihydrofluorescein diacetate (DCF-DA) assay. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and western blotting were performed to quantify the mRNA and protein expression levels, resepectiviely, of stress or apoptosis-related genes including p21, p53, heat shock protein 70 (Hsp70), heat shock protein 90 (Hsp90), Bax, Bcl-2, and caspase-3. The results of the MTT assay showed that 50 ㎍/㎖ SCE did not affect cell viability. ROS generation in mouse spermatocyte cells increased by treatment with 100 μM styrene, and decreased by co-treatment with SCE. SCE repressed the mRNA expression of stress-related genes, which increased by styrene treatment. In addition, SCE inhibited the apoptosis of mouse spermatocyte cells by ameliorating mRNA and protein levels of apoptotic genes that were altered by styrene treatment.
Conclusions: These results suggest that SCE may alleviate styrene toxicity in mouse spermatocyte cells by reducing ROS stress and regulating genes related to styrene toxicity.
재료 및 방법
1. 세포주 배양
2. 추출물 제조
4. 활성산소 (reactive oxygen species, ROS) 생성량 측정
5. Semi-quantitative reverse transcription polymerasechain reaction (RT-PCR)
6. 단백질 발현 분석
7. 통계 처리
결과 및 고찰
1. 마우스 정모세포주에서 스티렌과 삼백초 추출물의 처리농도 결정
2. 스티렌에 의한 활성산소종 생성에 삼백초 추출물이 미치는 영향 확인
3. 스티렌에 의한 유전자 발현에 삼백초 추출물이 미치는영향 확인
4. 스티렌에 의한 단백질 발현에 삼백초 추출물이 미치는영향 확인
감사의 글
REFERENCES
