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Toxicity evaluation of the mycotoxins, citrinin and ochratoxin A, using several animal cell lines
N Kitabatake1, E Doi, A B Trivedi
1Research Institute of Food Science, Kyoto University, Japan.
Abstract:
1. The cytotoxicities of the nephrotoxic mycotoxins, citrinin and ochratoxin A were assayed on HeLa, C3H/10T1/2, NIH/3T3, MDCK (canine kidney), and HeLa P3 cell lines, using the MTT colorimetric assay. 2. Citrinin was less toxic than ochratoxin A in all of the cell lines examined. 3. The MDCK cells were more susceptible to both citrinin and ochratoxin A, in comparison with other cell lines. 4. Dose-responses, as measured by activities of leucine aminopeptidase and alkaline phosphatase of MDCK cells, were less sensitive than MTT colorimetric assay, indicating that these enzymes were not specifically inhibited in MDCK cells. 5. The LD50 of both toxins, calculated at 72 hr incubation, was in the same order as those reported from animal experiments using rats and mice.
Insights
Nephrotoxic mycotoxins citrinin and ochratoxin A showed varying cytotoxicities across cell lines. Canine kidney (MDCK) cells were most susceptible, with toxicity levels mirroring animal studies.
Area of Science:
- Toxicology
- Cell Biology
- Mycotoxicology
Background:
- Mycotoxins, such as citrinin and ochratoxin A, are toxic secondary metabolites produced by fungi.
- These nephrotoxic compounds pose risks to human and animal health, necessitating detailed toxicological studies.
- Understanding the differential cellular responses to these mycotoxins is crucial for risk assessment.
Purpose of the Study:
- To compare the cytotoxic effects of citrinin and ochratoxin A on various mammalian cell lines.
- To identify cell lines exhibiting differential sensitivity to these nephrotoxic mycotoxins.
- To evaluate the utility of the MTT assay versus enzyme activity measurements for assessing mycotoxin toxicity.
Main Methods:
- Cytotoxicity assays using the MTT colorimetric method were performed on HeLa, C3H/10T1/2, NIH/3T3, MDCK, and HeLa P3 cell lines.
- Leucine aminopeptidase and alkaline phosphatase activities were measured in MDCK cells to assess dose-response.
- The median lethal dose (LD50) for both mycotoxins was calculated after 72-hour incubation.
Main Results:
- Ochratoxin A exhibited higher cytotoxicity than citrinin across all tested cell lines.
- MDCK (canine kidney) cells demonstrated significantly greater susceptibility to both citrinin and ochratoxin A compared to other cell lines.
- The MTT assay proved more sensitive for detecting toxicity than measurements of leucine aminopeptidase and alkaline phosphatase activity in MDCK cells.
- Calculated LD50 values at 72 hours correlated with findings from in vivo animal studies.
Conclusions:
- MDCK cells are a sensitive model for evaluating the nephrotoxicity of citrinin and ochratoxin A.
- The MTT assay is a reliable method for assessing the cytotoxicity of these mycotoxins.
- The in vitro findings align with in vivo toxicity data, supporting the relevance of this cell-based assay for mycotoxin risk assessment.