Cardiovascular and Pulmonary PharmacologyParacetamol (acetaminophen) cytotoxicity in rat type II pneumocytes and alveolar macrophages In Vitro
Section snippets
Chemicals and materials
Acetaminophen, indomethacin, trypsin type I (EC 3.4.21.4, Cat. No. T-8003), GSH, pyruvic acid, NADH, metaphosphoric acid (HPO3), bovine serum albumin, Percoll, EDTA, MTT, DMSO, and o-phthaldialdehyde were purchased from Sigma. DMEM, FBS, penicillin–streptomycin solution (10,000 U/mL and 10,000 μg/mL, respectively), fungizone (250 μg/mL), and l-glutamine (200 mM) were obtained from GIBCO. DNase I (EC 3.1.21.1) was purchased from Boehringer Mannheim GmbH. Protein assay dye solution was obtained
In vitro toxicity of APAP
After 20-hr incubation, APAP caused a concentration-related decrease in MTT reduction in rat type II pneumocytes (freshly isolated and 24-hr-old) and alveolar macrophages (Fig. 1). The reduction of MTT was significantly decreased compared to control at ≥ 5 mM APAP in freshly isolated type II pneumocytes and alveolar macrophages, and at ≥ 10 mM in 24-hr-old type II cells. APAP caused a significant decrease in LDH retention in the fraction of attached cells in freshly isolated type II
Discussion
These studies demonstrated that rat type II pneumocytes and alveolar macrophages are sensitive to APAP-induced toxicity in vitro in a concentration-dependent manner, with the order of susceptibility being as follows: freshly isolated type II pneumocytes > alveolar macrophages > 24-hr-old type II pneumocytes. The concentrations that caused cytotoxicity may appear quite high, since APAP concentrations of ≥ 5 mM were needed to produce cytotoxicity in freshly isolated type II pneumocytes and
Acknowledgements
This work was supported by the Belgian Office for Scientific, Technical, and Cultural Affairs and partly by INCO/Copernicus (EU) (IC15-CT96-0314).
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