[Oxidative stress induced by NaAsO2 in HaCaT cells].
Xiance Sun, Guifan Sun, Shan Liu, Ying Zhang
Abstract
Xiance Sun, Guifan Sun, Shan Liu, Ying Zhang
Abstract
OBJECTIVE: To study the level of oxidative stress induced by sodium arsenite (NaAsO2) in HaCaT cells. METHODS: The AlamarBlue assay was used to evaluate the viability of HaCaT. The level of ROS was detected by staining. cells with DCFH-DA. The contents of reduced (GSH) and oxidated (GSSG) glutathione were detected with the fluorescent method. The apoptosis and necrosis rates were counted based on the PI staining. RESULTS: The reduction of AlamarBlue increased in the cells treated at dose of 0.001 -1 micromol/L NaAsO2 and decreased in the cells treated at doses of over 10 micromol/L NaAsO2. The fluorescent density of DCF significantly increased in all experimental groups, the contents of GSH and GSSG increased in groups higher than that of 1 micromol/L and higher than that of 5 micromol/L respectively. The apoptosis and necrosis rates were increased markedly at dose of 20 micromol/L. CONCLUSION: Arsenic could induce enhancive ROS in HaCaT. At low levels of arsenic the proliferation of HaCaT was stimulated while at high levels of arsenic was inhibited. The increase in contents of GSH and GSSG could be associated with the excessive ROS and with detoxification in HaCaT.
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OBJECTIVE: To study the level of oxidative stress induced by sodium arsenite (NaAsO2) in HaCaT cells. METHODS: The AlamarBlue assay was used to evaluate the viability of HaCaT. The level of ROS was detected by staining. cells with DCFH-DA. The contents of reduced (GSH) and oxidated (GSSG) glutathione were detected with the fluorescent method. The apoptosis and necrosis rates were counted based on the PI staining. RESULTS: The reduction of AlamarBlue increased in the cells treated at dose of 0.001 -1 micromol/L NaAsO2 and decreased in the cells treated at doses of over 10 micromol/L NaAsO2. The fluorescent density of DCF significantly increased in all experimental groups, the contents of GSH and GSSG increased in groups higher than that of 1 micromol/L and higher than that of 5 micromol/L respectively. The apoptosis and necrosis rates were increased markedly at dose of 20 micromol/L. CONCLUSION: Arsenic could induce enhancive ROS in HaCaT. At low levels of arsenic the proliferation of HaCaT was stimulated while at high levels of arsenic was inhibited. The increase in contents of GSH and GSSG could be associated with the excessive ROS and with detoxification in HaCaT.
Key concepts: HaCaT, Sodium arsenite, Glutathione, Oxidative stress, Apoptosis, Molecular biology, Chemistry, Staining