Objective To study the effects of resveratrol on hypoxia injury in rat islet beta tumor cells (INS-1). Methods The INS-1 cells were divided into three groups: control group, hypoxia group, hypoxia and resveratrol group (hypoxia+RSV). The cell proliferation at different hypoxia times (6, 12, 24, 48 h) was detected by CCK8 kit. Resveratrol at the concentrations of 8, 10, 12, 15 μmol/L were used for intervention for 12 hours. Oxidative stress levels were detected by reactive oxygen species kit and mitochondrial superoxide kit. Mitochondrial membrane potential and ATP levels were detected for mitochondrial function, and apoptosis was detected by flow cytometry. Insulin quantitative assay kit was used to detect insulin secretion, and quantitative real-time reverse transcription was used to detect the mRNA levels of Sirt1, Pdx1 and Glut2 genes. The corresponding protein levels were detected by Western blot. Results Compared with the normal oxygen control group, cell proliferation was decreased (P<0.05), apoptosis rate increased (P<0.01), mitochondrial function decreased (P<0.05), and insulin secretion decreased (P<0.05) in the hypoxia group. The expression levels of Sirt1, Pdx1, Glut2 genes and their corresponding proteins were also decreased (P<0.05). The intervention by resveratrol could significantly reduce hypoxia stress induced damage (P<0.05), promote proliferation (P<0.05), decrease apoptosis (P<0.01), improve mitochondrial function (P<0.05), and promote insulin secretion (P<0.05) in INS-1 cells exposed to hypoxia. The expression levels of Sirt1, Pdx1, Glut2 genes and their corresponding proteins were also improved in response to resveratrol intervention (P<0.05). Conclusion Resveratrol can protect INS-1 cells against oxidative stress caused by hypoxia, improve mitochondrial function, inhibit apoptosis, promote insulin function gene expression and increase insulin secretion.
Key words
resveratrol /
hypoxia /
INS-1 cells
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References
[1] Wilson DF, Matschinsky FM.Oxygen dependence of glucose sensing: role in glucose homeostasis and related pathology[J].J Appl Physiol,2019,126:1746–1755.
[2] Cavallari G, Olivi E, Bianchi F, et al. Mesenchymal stem cells and islet cotransplantation in diabetic rats: improved islet graft revascularization and function by human adipose tissue-derived stem cells preconditioned with natural molecules[J]. Cell Transplant, 2012, 21: 2771–2781.
[3] Stancill JS, Corbett JA.The role of thioredoxin/peroxi-redoxin in the β-cell defense against oxidative damage[J]. Front Endocrinol (Lausanne),2021,12:718235.
[4] Roma LP, Jonas JC.Nutrient metabolism, subcellular redox state, and oxidative stress in pancreatic islets and β-cells[J].J Mol Biol,2020,432:1461–1493.
[5] Meng Q, Li J, Wang C, et al. Biological function of resveratrol and its application in animal production: a review[J].J Anim Sci Biotechnol,2023,14:25.
[6] Li S, Feng F, Deng Y.Resveratrol regulates glucose and lipid metabolism in diabetic rats by inhibition of PDK1/AKT phosphorylation and HIF-1α expression[J]. Diabetes Metab Syndr Obes,2023,16:1063–1074.
[7] Benjamin JS, Culpepper CB, Brown LD, et al. Chronic anemic hypoxemia attenuates glucose-stimulated insulin secretion in fetal sheep[J]. Am J Physiol Regul Integr Comp Physiol,2017,312:R492–R500.
[8] Pae EK, Ahuja B, Kim M, et al. Impaired glucose homeostasis after a transient intermittent hypoxic exposure in neonatal rats[J]. Biochem Biophys Res Commun,2013,441:637–642.
[9] Uchiyama T, Ota H, Ohbayashi C, et al. Effects of intermittent hypoxia on cytokine expression involved in insulin resistance[J]. Int J Mol Sci,2021,22: 12898.
[10] Gao T, McKenna B, Li C, et al. Pdx1 maintains β cell identity and function by repressing an α cell program[J]. Cell Metab,2014,19:259–271.
[11] Kaneto H, Xu G, Fujii N, et al. Involvement of c-Jun N-terminal kinase in oxidative stress-mediated suppression of insulin gene expression[J]. J Biol Chem,2002,277:30010–30018.
[12] Matsuoka T, Kajimoto Y, Watada H, et al. Glycation-dependent, reactive oxygen species-mediated suppres-sion of the insulin gene promoter activity in HIT cells[J]. J Clin Invest,1997,99:144–150.
[13] Leenders F, Groen N, de Graaf N, et al. Oxidative stress leads to β-cell dysfunction through loss of β-cell identity[J]. Front Immunol,2021,12:690379.
[14] Wang RH, Xu X, Kim HS, et al. SIRT1 deacetylates FOXA2 and is critical for Pdx1 transcription and β-cell formation[J]. Int J Biol Sci,2013,9:934–946.
[15] Giuliani M, Moritz W, Bodmer E, et al. Central necrosis in isolated hypoxic human pancreatic islets: evidence for postisolation ischemia[J]. Cell Transplant,2005,14: 67–76.
