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txt Background In this investigation, we explored the potential cytotoxic interaction between LPS and CEES using a murine macrophage cell line (RAW264.
Both bis-2-(chloroethyl)sulfide) and CEES are known to provoke acute inflammatory responses in skin [ 1 2 3 ] .
Apoptosis is now considered a possible molecular mechanism whereby CEES induces cytotoxicity [ 5 6 ] .
In this study, we also determined if phorbol myristate acetate (PMA) activation of PKC also enhanced CEES toxicity.
Results Cytotoxic interaction between LPS and CEES as measured by the MTT assay RAW 264.
05) to CEES toxicity (24 hr) than resting macrophages as indicated by the dramatic drop in dehydrogenase activity.
The characteristics of CEES toxicity to LPS-stimulated RAW 264.
However, LPS level at a level of 100 ng/ml significantly increased the cytotoxicity of 300 µM CEES compared to LPS activated RAW265.
Statistical analyses indicate that cells treated with 100 µM CEES were not significantly different from control cells (not treated with CEES) in the presence of 0, 25 or 50 ng/ml of LPS.
However, at a LPS level of 100 ng/ml, cells treated with 100 µM CEES showed a great MTT optical density than control cells.
Time course for the cytotoxic interaction between LPS and CEES as measured by the propidium iodide (PI) assay In order to insure that our results could be reproduced with an alternative assay for cell viability, we also utilized the propidium iodide (PI) assay.
Moreover, the cytotoxic interaction between LPS and CEES was noticeable after 6 hours (p < 0.
TNF-alpha and IL-1beta also enhance the cytotoxic effect of CEES LPS from Gram-negative bacteria binds to CD14 and initiates a complex signal transduction pathway involving the Toll receptor family, which eventually results in the synthesis of pro-inflammatory cytokines such as TNF-alpha and IL-1beta.
We, therefore, investigated the potential roles of TNF-alpha and IL-1beta in enhancing CEES cytotoxicity.
, an amplification of CEES toxicity (after 24 hours) in the presence of the inflammatory cytokine.
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