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- Title
Protective effects of γ-aminobutyric acid against H<sub>2</sub>O<sub>2</sub>-induced oxidative stress in RIN-m5F pancreatic cells.
- Authors
Tang, Xue; Yu, Renqiang; Zhou, Qin; Jiang, Shanyu; Le, Guowei
- Abstract
Background: γ-Aminobutyric acid (GABA) is a major inhibitory neurotransmitter in the central nervous system and reported to maintain the redox homeostasis and insulin secretion function of pancreatic β cells. This study tested the hypothesis that GABA maintains cellular redox status, and modulates glycogen synthase kinase (GSK)-3β and antioxidant-related nuclear factor erythroid 2-related factor 2 (NRF2) nuclear mass ratio in the H2O2-injured RINm5F cells. Methods: RINm5F cells were treated with/without GABA (50, 100 and 200 μmol/L) for 48 h and then exposed to 100 μmol/L H2O2 for 30 min. Viable cells were harvested, and dichloro-dihydro-fluorescein diacetate (DCFH-DA) was used to detect reactive oxygen species (ROS) level; cellular redox status and insulin secretion were measured; cell viability was determined by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay; mitochondrial membrane potential (MMP) was detected by flow cytometry; relative genes levels were analyzed by reverse transcriptase polymerase chain reaction (RT-PCR); western blotting was used to determine protein expression of GSK-3β and p-GSK-3β (Ser9), and nuclear and cytoplasmic NRF2. Results: H2O2 increased ROS production, and induced adverse affects in relation to antioxidant defense systems and insulin secretion. These changes were restored by treatment with 100 and 200 μmol/L GABA. In addition, 100 or 200 μmol/L GABA induced membrane depolarization and increased cell viability. These effects were mediated by Caspase-3, Bcl-2 associated X protein (Bax) and B-cell lymphoma-2 (Bcl-2) expression. Western blotting indicated that GABA inhibited GSK-3β by increasing p-GSK-3β (Ser9) level, and directed the transcription factor NRF2 to the nucleus. Conclusion: In rat insulin-producing RINm5F cells, GABA exerts its protective effect by regulating GSK-3β and NRF2, which governs redox homeostasis by inhibiting apoptosis and abnormal insulin secretion by exposure to H2O2.
- Subjects
REACTIVE oxygen species; ANIMAL experimentation; APOPTOSIS; BIOLOGICAL transport; FLOW cytometry; GABA; GENE expression; HOMEOSTASIS; HYDROGEN peroxide; INSULIN; ISLANDS of Langerhans; ONCOGENES; OXIDATION-reduction reaction; POLYMERASE chain reaction; PROTEIN kinases; RATS; TRANSCRIPTION factors; WESTERN immunoblotting; OXIDATIVE stress; REVERSE transcriptase polymerase chain reaction
- Publication
Nutrition & Metabolism, 2018, Vol 15, Issue 1, pN.PAG
- ISSN
1743-7075
- Publication type
Article
- DOI
10.1186/s12986-018-0299-2