Eicosapentaenoic Acid and γ-Linolenic Acid Induce Apoptosis in HL-60 Cells

• Published in: The Journal of surgical research 2002-09, Vol.107 (1), p.145-153
• Main Authors: Gillis, R.C., Daley, B.J., Enderson, B.L., Karlstad, M.D.
• Format: Article
• Language: English
• Subjects: Apoptosis - drug effects; ARDS; Biological and medical sciences; borage oil; cell culture; Cell Survival - drug effects; DNA Fragmentation - drug effects; Dose-Response Relationship, Drug; Drug Combinations; Eicosapentaenoic Acid - administration & dosage; Eicosapentaenoic Acid - pharmacology; fish oil; Flow Cytometry; gamma-Linolenic Acid - administration & dosage; gamma-Linolenic Acid - pharmacology; HL-60 Cells - drug effects; HL-60 Cells - physiology; Humans; inflammation; Medical sciences; necrosis; neutrophil; Pneumology; polyunsaturated fatty acids; pulmonary; Respiratory system : syndromes and miscellaneous diseases; sepsis
• ISSN: 0022-4804; 1095-8673
• DOI: 10.1006/jsre.2002.6496

Enteral nutrition with eicosapentaenoic acid (EPA; 20:5 n-3) and γ-linolenic acid (GLA; 18:3 n-6) decreased pulmonary inflammation by reducing neutrophil counts and chemotactic factors in bronchoalveolar lavage fluid during acute respiratory distress syndrome (ARDS). We hypothesize that the anti-inflammatory effects of EPA and GLA may be due, in part, to induction of neutrophil apoptosis. The purpose of this study was to determine whether EPA and GLA, alone or in combination, trigger apoptotic cell death in the human promyelocytic leukemia HL-60 cell line. HL-60 cells were incubated with 10, 20, 50, and 100 μmol/L EPA, GLA, or various combinations of EPA and GLA for 2, 4, 8, 12, and 24 hs. Oleic acid (18:1 n-9) was used as a fatty acid control. Flow cytometry using dual staining with propidium iodide and annexin V–FITC assessed apoptosis, necrosis, and viability. Apoptosis was verified by DNA fragmentation as assessed by agarose gel electrophoresis. EPA, GLA, and various combinations of EPA and GLA significantly induced apoptosis and reduced cell viability in HL-60 cells. Viability was significantly reduced to the same extent with the combination of 50 μmol/L EPA\20 μmol/L GLA compared with 100 μmol/L EPA. These data indicate that EPA and GLA, alone or in combination, reduce cell survival by induction of apoptosis. Thus, induction of apoptosis by select dietary n-3 (EPA) and n-6 (GLA) polyunsaturated fatty acids may be the mechanism of the resolution of pulmonary inflammation in ARDS.