Docosahexaenoic acid attenuates oxidative stress and protects human gingival fibroblasts against cytotoxicity induced by hydrogen peroxide and butyric acid
| Autor: | Anna Janas, Emilia Zgórzyńska, Barbara Dziedzic, Anna Walczewska, Monika Witusik-Perkowska, Anita Wierzbicka-Ferszt, Anna Zwolinska |
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| Rok vydání: | 2015 |
| Předmět: |
Docosahexaenoic Acids
Cell Survival Apoptosis Biology medicine.disease_cause Gas Chromatography-Mass Spectrometry Butyric acid Necrosis chemistry.chemical_compound medicine Humans MTT assay Viability assay Propidium iodide General Dentistry Cells Cultured Membrane Potential Mitochondrial chemistry.chemical_classification Reactive oxygen species Fatty Acids Hydrogen Peroxide Cell Biology General Medicine Fibroblasts Respiratory burst Oxidative Stress Otorhinolaryngology Biochemistry chemistry Docosahexaenoic acid Butyric Acid Reactive Oxygen Species Oxidative stress |
| Zdroj: | Archives of Oral Biology. 60:144-153 |
| ISSN: | 0003-9969 |
| DOI: | 10.1016/j.archoralbio.2014.09.009 |
| Popis: | Objective The oxidative burst of the host cells associated with bacterial pathogen infection contributes to the destruction of periodontal tissue. The present study investigates the effect of docosahexaenoic acid (DHA) on human gingival fibroblast (HGF) viability and ROS generation. Methods The cell viability by MTT assay, ROS level using H 2 DCF-DA probe, and protein thiol content were measured in HGFs after 24 h preincubation with different concentrations of DHA followed by treatment with H 2 O 2 . The cell death rate was determined by Annexin V/propidium iodide staining, and mitochondrial membrane potential (Δ Ψm ) was examined by MitoTracker Red probe in H 2 O 2 - and butyric acid-treated HGFs. The fatty acid composition of plasma membranes after incubation with DHA was determined by gas chromatography mass spectrometry. Results DHA preincubation in a dose-dependent manner increased the viability of HGFs exposed to H 2 O 2 and decreased ROS generation compared to the control cells. In HGFs preincubated with 30 μM DHA, the Δ Ψm significantly increased in both H 2 O 2 - and butyric acid-treated cells. Moreover, incubation with DHA preserved the protein thiol level as effectively as N-acetylcysteine. Application of 50 μM DHA increased the quantity of viable cells, decreased the number of necrotic cells after H 2 O 2 treatment, and protected HGFs from apoptosis induced by butyric acid. DHA in the plasma membranes of these HGFs represented about 6% of the total amount of fatty acids. Conclusions These results demonstrate that enrichment of HGFs with DHA reduces ROS generation and enhances the mitochondrial membrane potential protecting the fibroblasts against cytotoxic factors. |
| Databáze: | OpenAIRE |
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