Centaurium erythraea Methanol Extract Attenuates SNP-lnduced Oxidative Stress in Pancreatic B-Cells
2018
Authors:
Đorđević, MilošMihailović, Mirjana
Arambašić Jovanović, Jelena
Grdović, Nevena
Uskoković, Aleksandra
Đorđević, Marija
Rajić, Jovana
Tolić, Anja
Poznanović, Goran
Vidaković, Melita
Dinić, Svetlana
Document Type:
Conference object (Published version)
,
© 2018 Silverchair Publisher
Metadata
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Objectives: Centaurium erythraea ( CE) is traditionally used in Serbia for diabetes management. Since oxidative stress represents one of the major pathogenic factors that lead to diabetes and its complications, this study investigated protective effect of CE methanol extract against oxidative stress-induced pancreatic P-cell death.
Methods: Rin-SF, rat insulinoma pancreatic P-cells, were incubated for 24 h with 1.25 mM sodium nitroprusside (SNP) with/ without CE extract (0.2 mg/mL) and processed immediately. RinSF cell viability was estimated using the MTT viability assay. Lipid peroxidation was assessed using the thiobarbituric acid-reactive substance (TBARS) assay, while the DNA damage was estimated by alkaline comet assay. Catalase (CAT) activity was determined by the rate ofH202 decomposition, whereas superoxide dismutase (SOD) activity was estimated by the epinephrine method. The activities of glutathione peroxidase (GPx) and glutathione reductase ( GR) were determined by monitoring NADPH oxidation. Relative gene expression of CAT, MnSOD, CuZnSOD, GPx, GR and insulin was determined by RT-qPCR. Relative protein level of antioxidant enzymes was estimated using immunoblot analysis.
Results: CE methanol extract enhanced p-cell viability and insulin gene expression by reducing oxidative stress in SNP-treated cells. CE extract lowered DNA damage and lipid peroxidation provoked by SNP treatment and adjusted antioxidant enzyme activities. CE treatment increased SNP-mediated attenuation of CAT, GPx and GR activities and reduced CuZnSOD and MnSOD activities that were stimulated in SNP treated cells. SNP-induced increase in gene expression of CAT, GPx, GR, MnSOD and CuZnSOD was accompanied by decrease of CAT, GPx and CuZnSOD mRNA level after CE treatment. In addition, SNP treatment increased protein levels of CAT, GR, GPx and MnSOD and decreased CuZnSO D protein level. CE extract reduced CAT and MnSOD and partially restored CuZnSOD protein level.
Conclusions: The CE methanol extract protects pancreatic p-cells from oxidative damage by improving antioxidant defence system. Detected attenuation of oxidative stress in P-cells in vitro provide a useful platform for in vivo investigation of antioxidant and antidiabetic effect of CE extract and its potential usage as an effective supplement for diabetes treatment.
Funding / projects:
- European Cooperation in Science and Technology (COST) CM1406
- Signaling molecules in diabetes: search for potential targets in intrinsic pathways for prediction and intervention in diabetes (RS-MESTD-Basic Research (BR or ON)-173020)
In:
- Selected Abstracts from the 3rd European Summer School on Nutrigenomics; 2018 Jun 25-29; Jesi, Italy. Karger Publishers; 2018. p. 7. (Lifestyle Genomics; Vol. 11; No. 1).