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Comparative cytotoxicity of alachlor on RTG-2 trout and SH-SY5Y human cells
1Laboratory of Toxicology, Faculty of Pharmacy, University of Valencia, Av. Vicent Andrés Estellés s/n, 46100, Burjassot, València, Spain. monica.fernandez@uv.es
Abstract:
The cytotoxic effects of the herbicide alachlor were compared on rainbow trout gonadal RTG-2 and human neuroblastoma SH-SY5Y cell lines. The end points evaluated in both cells after 24, 48, and 72 h of exposure were total protein content (PC), lysosomal function, and mitochondrial's integrity by mitochondrial succinate dehydrogenase (SDH) activity. After 24 h, cytoplasmic membrane integrity by cytosolic lactate dehydrogenase (LDH) leakage and LDH intracellular activity were also studied. In addition, acetylcholinesterase activity (AChE) was quantified in SH-SY5Y cells. The possible biotransformation of alachlor by RTG-2 cells was investigated by analyzing the exposure culture medium by liquid chromatography-mass spectrometry. In RTG-2, EC50 values on PC, lysosomal function, and SDH activity after 24 h exposure ranged from 80 to 95 microM and decreased to approximately 40 microM for longer exposure time periods. SH-SY5Y cells were slightly more sensitive than RTG-2 cells, with EC50 values on PC and lysosomal function ranging from 87 to 75 microM at 24 h and decreasing to 47 microM and 34 microM at 72 h, respectively. AChE activity was increased, being the most sensitive marker for SH-SY5Y with an EC50 of 20 microM at 24 h. The metabolic enzyme SDH was stimulated in SH-SY5Y and reduced in RTG-2 cells. At the studied conditions, no metabolites of alachlor were detected in RTG-2 cultures. In conclusion, the proposed battery approach is an effective screening tool for the safety assessment of environmental contaminants as a complement to fish and animal toxicity procedures.
Insights
This study assessed the cytotoxic effects of alachlor on rainbow trout (RTG-2) and human (SH-SY5Y) cells, finding it toxic to both. The results support using a cell-based assay for environmental contaminant safety assessments.
Area of Science:
- Environmental toxicology
- Cell biology
- Biomarker discovery
Background:
- Alachlor is a widely used herbicide with potential environmental risks.
- Assessing herbicide toxicity using cell lines can complement traditional animal testing.
Purpose of the Study:
- To compare the cytotoxic effects of alachlor on rainbow trout (RTG-2) and human (SH-SY5Y) cell lines.
- To evaluate various cellular endpoints as potential biomarkers for alachlor toxicity.
- To investigate the potential for alachlor biotransformation in RTG-2 cells.
Main Methods:
- Exposure of RTG-2 and SH-SY5Y cells to alachlor for 24, 48, and 72 hours.
- Measurement of total protein content, lysosomal function, mitochondrial succinate dehydrogenase (SDH) activity, and lactate dehydrogenase (LDH) leakage.
- Quantification of acetylcholinesterase (AChE) activity in SH-SY5Y cells.
- Analysis of culture medium for alachlor metabolites using liquid chromatography-mass spectrometry.
Main Results:
- Alachlor exhibited dose-dependent cytotoxicity in both cell lines, with EC50 values decreasing over time.
- SH-SY5Y cells were generally more sensitive to alachlor than RTG-2 cells.
- AChE activity in SH-SY5Y cells was the most sensitive biomarker, with an EC50 of 20 microM at 24 hours.
- SDH activity was stimulated in SH-SY5Y cells but reduced in RTG-2 cells.
- No alachlor metabolites were detected in RTG-2 cell cultures under the studied conditions.
Conclusions:
- The study demonstrates the differential sensitivity of RTG-2 and SH-SY5Y cells to alachlor.
- A battery of cellular assays, including AChE activity, can serve as an effective screening tool for environmental contaminant safety.
- This cell-based approach can complement traditional fish and animal toxicity testing methods.
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