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Effect of hexavalent chromium-treated sperm on <i>in vitro</i> fertilization and embryo development
Yoisungnern, Ton,Das, Joydeep,Choi, Yun-Jung,Parnpai, Rangsun,Kim, Jin-Hoi PRINCETON SCIENTIFIC PUBLISHING CO 2016 TOXICOLOGY AND INDUSTRIAL HEALTH Vol.32 No.9
<P>Hexavalent chromium (Cr(VI)) is an environmental contaminant that is associated with reproductive abnormalities in both humans and animals. In the present study, we evaluated the cytotoxic effect of Cr(VI) on sperm function and subsequent embryo development after in vitro fertilization (IVF). Sperm obtained from BDF1 male mice were treated with potassium dichromate (0, 3. 125, 6.25, 12.5, 25, or 50 mu/M) for 3 h. Cr(VI) significantly decreased sperm viability and acrosome reaction with increasing dose. These Cr(VI)-treated sperms were further used for IVF of oocytes obtained from BDF1 female mice. Results showed that Cr(VI)treated sperm caused a significant reduction in IVF success, higher developmental arrest at the two-cell stage of embryos, and delayed blastocyst formation with increasing dose. In particular, most blastocysts from the Cr(VI)-treated sperm resulted in hatching failure as well as decreased inner cell mass and trophectoderm (TE). Furthermore, blastocysts obtained from Cr(VI)-treated sperm showed lower expression of not only TE-associated genes (eomes, cdx2, and krt8) but also pluripotent marker genes (sox2, pou5fl, and klf4) that are responsible for further embryo development of blastocyst embryos. The results of our current study showed that Cr(VI)-treated sperm had negative effects on oocyte fertilization and subsequent embryo development.</P>
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Diffusion flame-derived fine particulate matters doped with iron caused genotoxicity in B6C3F1 mice
Park, Jin Hong,Han, Kyu Tae,Eu, Kook-Jong,Kim, Jun-Sung,Chung, Kyu Hyuk,Park, Bio,Yang, Go Su,Lee, Kee-Ho,Cho, Myung-Haing PRINCETON SCIENTIFIC PUBLISHING CO 2005 TOXICOLOGY AND INDUSTRIAL HEALTH Vol.21 No.3-4
<P>Potential genotoxic effects of diffusion flame-derived particulate matters (PMs), known to cause various adverse health problems, doped with iron, one of the representative heavy metals frequently found in the atmosphere, were examined. B6C3F1 mice were exposed to PMs [chamber 1 (low), 100; chamber 2 (middle), 200; and chamber 3 (high), 400 microg/m3] for 6 h/day, 5 days/week for one, two and four weeks in 1.5 m3 whole-body inhalation chambers. Our diffusion flame system produced 94.8 and 5.2% fine PM2.5 and PM10, respectively, with 89% of PM2.5 sized between 0.1 and 0.2 microm. Two cytogenetic endpoints were investigated through chromosomal aberration and supravital micronucleus (SMN) assays. Frequencies of cells with chromosome aberration (%) were observed in time- and concentration-dependent manners except in one-week exposure group, as also observed in SMN study. Generally, noniron flame induced less chromosome aberration than iron-doped flame, an indication that iron particles could potentiate urban PM toxicity. The above results indicate our diffusion flame system generated genotoxic fine PMs, whose effects were potentiated by organometallic particles such as iron. Our system can provide reliable PM models for studying the toxicity of urban fine PMs applicable for risk assessment.</P>
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Kwon, Mi-Jeong,Kang, Seok-Jin,Park, Young-Il,Yang, Yool-Hee,Bang, Sa-Ik,Park, Yong Ho,So, ByungJae,Cho, Myung-Haing,Kang, Hwan-Goo Princeton Scientific Publishers 2015 Cell biology and toxicology Vol.31 No.3
<P>In the present study, we differentiated hepatocyte-like cells (HLCs) from human adipose tissue-derived mesenchymal stem cells (AT-MSCs). The hepatic differentiation was confirmed by increases in hepatic proteins or genes, the cytochrome P450 (CYP) activities, albumin secretion, and glycogen storage. To determine the developmental toxic effect of arsanilic acid (Ars) and acetaminophen (AAP) on the hepatic development, the differentiating cells were treated with the test chemicals (below IC12.5) from day 4 to day 13. The enzymatic activities of lactate dehydrogenase (LDH), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) did not significantly differ in response to Ars treatment. AAP treatment increased the activities of all enzymes in a dose-dependent manner, significantly at concentrations of 2.5 and 5 mM of AAP. On the expressions of hepatic genes for Ars, the expressions were significantly inhibited by more than 0.5 mM for Albumin (ALB), but only 2.5 mM for α-feto protein (AFP). In the AAP-treated group, the expressions of ALB and AFP were significantly decreased at the concentrations exceeding 0.625 mM. The activities of CYP3A4 were not changed by both treatments. The activities of CYP1A2 were increased by AAP, whereas it was decreased by Ars treatment. In conclusion, AAP could cause serious adverse effects during the hepatic development as compared to Ars.</P>
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Kang, Seok-Jin,Jeong, Sang-Hee,Kim, Eun-Joo,Cho, Joon-Hyoung,Park, Young-Il,Park, Sung-Won,Shin, Hyo-Sook,Son, Seong-Wan,Kang, Hwan-Goo Princeton Scientific Publishers 2013 Cell biology and toxicology Vol.29 No.1
<P>Embryonic stem cell testing is an alternative model system to assess drug and chemical toxicities because of its similar developmental characteristics with in vivo embryogenesis and organogenesis. This study evaluated the toxicity of chemicals at specific developmental stages of mouse embryonic stem cell (ESC)-derived hepatic differentiation; hepatic progenitor cells (HPCs), and hepatocyte-like cells (HCs). The toxic effects of carbon tetrachloride (CCl(4)), 5-fluorouracil (5-FU), and arsanilic acid (Ars) were evaluated by measuring the expressions of Cytokeratin (CK18) and GATA binding protein 4 (GATA-4) and the activities of aspartate transaminase (AST), lactate dehydrogenase (LDH), and alkaline phosphatase (ALP) during the hepatic differentiation process. Non-toxic doses of three chemicals at a range of 25 to 500?μM for CCl(4), 12.5 to 800?nM for 5-FU and 6.25 to 400?mM for Ars were treated. In the CCl(4)-treated group, significant decreases (P?<?0.05) of the marker expression were observed by more than 300?μM from day 10 in CK18 and by more than 400?μM of CCl(4) from day 22 in GATA-4, respectively. However, both markers were decreased (P?<?0.01) by treatments of all doses at day 40. In the 5-FU-treated group, the expressions of two proteins were not affected by any of the doses at day 10 and 22, whereas the GATA-4 expression was decreased (P?<?0.05) by more than 400?nM of 5-FU at days 28 and 40. In the Ars-treated group, the CK18 expression was inhibited (P?<?0.05) by more than 100?mM of Ars at day 22 but showed a tendency to recover. Although the GATA-4 was inhibited by all doses at day 22, the inhibition of GATA-4 recovered at days 28 and 40. ALP activities of three chemicals were significantly increased (P?<?0.05) by a dose-dependent manner. The activities of AST and LDH were prone to be increased by more than 300?μM of CCl(4,) but not affected by all doses of 5-FU except for 800?nM of 5-FU in AST activities. In the Ars, the enzyme activities were significantly increased (P?<?0.05) by more than 50?μM of Ars in AST and more than 6.25?μM of Ars in LDH. The present results indicate that CCl(4) has a more toxic effect on HCs, whereas Ars is more toxic to HPCs. Additionally, in vitro alternative testing using ESC-derived HPCs and HCs could provide useful information on chemical toxicity during the hepatic differentiation process and could be a useful model system for assessing chemical hepatotoxicity.</P>
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Lee, Tae-Bum,Moon, Young-Sook,Choi, Cheol-Hee Princeton Scientific Publishers 2012 Cell biology and toxicology Vol.28 No.1
<P>We explored if epigenetic mechanisms could be involved in the down-regulated expression of catalase gene (CAT) in the doxorubicin-resistant acute myelogenous leukemia (AML)-2/DX100 cells. Down-regulated CAT expression in AML-2/DX100 cells was completely recovered after treatment of hydrogen peroxide (H(2)O(2)) and histone deacetylase inhibitor, trichostatin A (TSA) but was increased slightly by the treatment of DNA methylation inhibitor, 5-aza-2'-deoxycytidine (5-AdC). Bisulfite-sequencing PCR revealed that a CpG island of CAT was not methylated in AML-2/DX100 cells. Chromatin immunoprecipitation assay confirmed that acetylation of histone H4 in AML-2/DX100 cells significantly decreased as compared with that in AML-2/WT cells, which was significantly increased by TSA more than 5-AdC. Meanwhile, overexpression of other up-regulated peroxidase genes appears to make compensation for decreased H(2)O(2)-scavenging activity for the down-regulated CAT expression in AML-2/DX100 cells. These results suggest that histone H4 deacetylation is responsible for the down-regulated CAT expression in AML-2/DX100 cells, which are well adapted to oxidative stress.</P>
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Kim, Ki Cheon,Lee, In Kyung,Kang, Kyoung Ah,Kim, Hye Sun,Kang, Sam Sik,Hyun, Jin Won Princeton Scientific Publishers 2012 Cell biology and toxicology Vol.28 No.6
<P>Oxidative stress caused by reactive oxygen species (ROS) induces DNA base modifications and DNA strand breaks. In this study, the protective effect of baicalein against H(2)O(2)-induced DNA damage was investigated in V79-4 Chinese hamster fibroblast cells. H(2)O(2) treatment increased the levels of intracellular ROS and DNA double-strand breaks (DSBs) and decreased the level of Ku70 protein and the phosphorylation (activation) of DNA-dependent protein kinase catalytic subunit (DNA-PKcs), which are involved in the repair of DSBs by nonhomologous end joining. Baicalein effectively scavenged intracellular ROS induced by H(2)O(2), reduced DSBs, and rescued Ku70 protein level and phosphorylation of DNA-PKcs. In cellular response to DNA base damage, 8-oxoguanine DNA glycosylase 1 (OGG1) plays a vital role in the removal of 8-oxoguanine (8-OxoG), which is formed mainly by oxidative stress. Baicalein significantly decreased the levels of 8-OxoG induced by H(2)O(2), and this correlated with increases in OGG1 promoter activity and OGG1 mRNA and protein expression. The phosphorylated form of Akt kinase, which is a regulator of OGG1, was sharply decreased by H(2)O(2), but was prevented by baicalein. A specific Akt inhibitor abolished the cytoprotective effects of baicalein, suggesting that OGG1 induction by baicalein involves the Akt pathway. In conclusion, baicalein exerted protective effects against DNA damage induced by oxidative stress by activating DNA repair systems and scavenging ROS.</P>
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