Signaling for ethanol-induced apoptosis and repair in vitro
| Autor: | Karen Valentino, Gady G. Katz, Neil H. Shear, Izabella M. Malkiewicz, Manuela G. Neuman |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
DNA Repair
Clinical Biochemistry Apoptosis Enzyme-Linked Immunosorbent Assay Caspase 3 Cell Line chemistry.chemical_compound In Situ Nick-End Labeling Humans Caspase Ethanol TUNEL assay biology General Medicine Molecular biology In vitro Hep G2 Microscopy Electron chemistry Caspases Hepatocytes biology.protein DNA fragmentation Signal Transduction |
| Zdroj: | Clinical Biochemistry. 34:219-227 |
| ISSN: | 0009-9120 |
| Popis: | Objectives: To evaluate whether caspases are involved in ethanol (EtOH)-induced apoptosis and if polyenylphosphatidylcholine (PPC) affects apoptosis, in vitro in Hep G2 cells. Methods: Cells were treated with 100 mmol/L EtOH for 24 h and with 2 doses of 100 mmol/L EtOH (1/24 h) in the presence of absence of 20 mmol/L of PPC or 50 μmol/L caspase 3 inhibitor (IDN). Cells were analyzed for apoptosis by transmission electron microscopy (TEM) 6000 cells/treatment, DNA fragmentation by ELISA and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (T dt-mediated d-UTP) nick-end-labeling, TUNEL. Results: 100 mmol/L dose of EtOH resulted in 22 ± 2.5% (p < 0.001) apoptosis (vs. control). Two consecutive doses of 100 mmol/L EtOH for 24 h each caused 36 ± 3.0% (p < 0.001 vs. control and p < 0.05 vs. one dose). PPC significantly reduced apoptosis (vs. non exposed to PPC): 100 mmol/L −12 ± 1.5% (p < 0.05) and 2 × 10−0 mmol/L −20 ± 2.0% (p < 0.001). Pretreatment with 50 μmol caspase inhibitor reduced EtOH-induced apoptosis in a similar proportion. Conclusions: PPC downregulates EtOH-apoptosis by a mechanism similar to caspase inhibition. |
| Databáze: | OpenAIRE |
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