Protective Effect of Saururus chinensis Ethanol Extract against Styrene in Mouse Spermatocyte Cell Line
2017
Yoon, J.H., NIHHS, RDA, Eumseong, Republic of Korea | Sohn, S.H., Croen Research Inc., Suwon, Republic of Kora | Lee, E.Y., NIHHS, RDA, Eumseong, Republic of Korea | Kim, G.S., NIHHS, RDA, Eumseong, Republic of Korea | Lee, S.E., NIHHS, RDA, Eumseong, Republic of Korea | Lee, D.Y., NIHHS, RDA, Eumseong, Republic of Korea | Seo, K.H., NIHHS, RDA, Eumseong, Republic of Korea | Lee, S.W., NIHHS, RDA, Eumseong, Republic of Korea | Kim, H.D., NIHHS, RDA, Eumseong, Republic of Korea
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 microgram/microliter 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.
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