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Chokeberry Anthocyanin Extract as Pancreatic β-Cell Protectors in Two Models of Induced Oxidative Stress.

Rugină D, Diaconeasa Z, Coman C, Bunea A, Socaciu C, Pintea A - Oxid Med Cell Longev (2015)

Bottom Line: The results showed that physiologically achievable concentrations of CAE (1, 5, and 10 μM) protect βTC3 against H2O2- and HG-induced cytotoxicity.Antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were increased in pancreatic β-cells pretreated with CAE compared to cells exposed to the prooxidant agents.These results demonstrate that anthocyanins from CAE were biologically active, showing a secretagogue potential and an antioxidative protection of enzymatic systems, conferring protection against H2O2 and glucose toxicity in βTC3 cells.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine, Mănăştur 3-5, 400372 Cluj-Napoca, Romania.

ABSTRACT
The aim of this study was to investigate the protective effects of a chokeberry anthocyanin extract (CAE) on pancreatic β-cells (βTC3) exposed to hydrogen peroxide- (H2O2-) and high glucose- (HG-) induced oxidative stress conditions. In order to quantify individual anthocyanins high performance liquid chromatography (HPLC) coupled to photodiode array (PDA) was used. The identification of the fragment ion pattern of anthocyanins was carried out by electrospray ionization mass spectrometry (LC-ESI-MS). The results showed that physiologically achievable concentrations of CAE (1, 5, and 10 μM) protect βTC3 against H2O2- and HG-induced cytotoxicity. Antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were increased in pancreatic β-cells pretreated with CAE compared to cells exposed to the prooxidant agents. GSH levels initially reduced after exposure to H2O2 and HG were restored by pretreatment with CAE. Insulin secretion in βTC3 cells was enhanced by CAE pretreatment. CAE restored the insulin pool and diminished the intracellular reactive oxygen species level in glucose-induced stress condition in βTC3 cells. These results demonstrate that anthocyanins from CAE were biologically active, showing a secretagogue potential and an antioxidative protection of enzymatic systems, conferring protection against H2O2 and glucose toxicity in βTC3 cells.

No MeSH data available.


Related in: MedlinePlus

Effect of CAE on insulin secretion and intracellular reactive oxygen species (ROS) level in βTC3 cells in stress-induced conditions. (a) HG-induced stress for 24 h and (b) H2O2-induced stress for 15 min. Data is expressed as mean ± SEM of two independent determinations. Statistical analyses: ∗P < 0.05 versus the control.
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fig5: Effect of CAE on insulin secretion and intracellular reactive oxygen species (ROS) level in βTC3 cells in stress-induced conditions. (a) HG-induced stress for 24 h and (b) H2O2-induced stress for 15 min. Data is expressed as mean ± SEM of two independent determinations. Statistical analyses: ∗P < 0.05 versus the control.

Mentions: In our study the administration of CAE concentrations of 1, 5, and 10 μM increased insulin secretion by 1, 9, and 18 ng of insulin/mg of protein, respectively, under basal glucose condition (Figure 5(b)(A)). The low dose of CAE (1 μM) increased the insulin secretion by 4 ng of insulin/mg of protein in HG (50 mM) stress condition (Figure 5(a)(A)). In a dose-dependent manner the CAE administration attempted to restore the pool of insulin in the HG (100 mM) environment (Figure 5(a)(A)). The CAE treatment in the presence of H2O2-induced stress did not have any influence on the secreted insulin in pancreatic β-cells (Figure 5(b)(A)). This may be possible if the insulin gene transcription was decreased after the exposure to H2O2 of pancreatic β-cells [41]. The insulin secretagogue effect of anthocyanins and anthocyanidins was demonstrated on pancreatic β-cells INS-1 832/13 previously [8]. In the same study an increment by the cyanidin-3-O-glucoside treatment of the insulin secretion was recorded, from 33 ng insulin/mg protein to 46 ng insulin/mg protein in basal glucose (4 mM) conditions. Glucose-induced stress (10 mM) increased for about 4 times the insulin secretion. It was observed that higher than 10 μg/mL (20 μM) cyanidin-3-O-glucoside concentrations did not significantly improve the insulin secretion in rodent pancreatic β-cells INS-1 832/13 [8]. Some in vivo studies reported that cyanidin-3-O-glucoside and anthocyanin-rich extracts ameliorated the hyperglycemia and the insulin sensitivity in diabetic mice [42, 43]. Insulin gene expression was decreased by exposure to oxidative stress in HIT-T15 cells or isolated rat islets [41].


Chokeberry Anthocyanin Extract as Pancreatic β-Cell Protectors in Two Models of Induced Oxidative Stress.

Rugină D, Diaconeasa Z, Coman C, Bunea A, Socaciu C, Pintea A - Oxid Med Cell Longev (2015)

Effect of CAE on insulin secretion and intracellular reactive oxygen species (ROS) level in βTC3 cells in stress-induced conditions. (a) HG-induced stress for 24 h and (b) H2O2-induced stress for 15 min. Data is expressed as mean ± SEM of two independent determinations. Statistical analyses: ∗P < 0.05 versus the control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig5: Effect of CAE on insulin secretion and intracellular reactive oxygen species (ROS) level in βTC3 cells in stress-induced conditions. (a) HG-induced stress for 24 h and (b) H2O2-induced stress for 15 min. Data is expressed as mean ± SEM of two independent determinations. Statistical analyses: ∗P < 0.05 versus the control.
Mentions: In our study the administration of CAE concentrations of 1, 5, and 10 μM increased insulin secretion by 1, 9, and 18 ng of insulin/mg of protein, respectively, under basal glucose condition (Figure 5(b)(A)). The low dose of CAE (1 μM) increased the insulin secretion by 4 ng of insulin/mg of protein in HG (50 mM) stress condition (Figure 5(a)(A)). In a dose-dependent manner the CAE administration attempted to restore the pool of insulin in the HG (100 mM) environment (Figure 5(a)(A)). The CAE treatment in the presence of H2O2-induced stress did not have any influence on the secreted insulin in pancreatic β-cells (Figure 5(b)(A)). This may be possible if the insulin gene transcription was decreased after the exposure to H2O2 of pancreatic β-cells [41]. The insulin secretagogue effect of anthocyanins and anthocyanidins was demonstrated on pancreatic β-cells INS-1 832/13 previously [8]. In the same study an increment by the cyanidin-3-O-glucoside treatment of the insulin secretion was recorded, from 33 ng insulin/mg protein to 46 ng insulin/mg protein in basal glucose (4 mM) conditions. Glucose-induced stress (10 mM) increased for about 4 times the insulin secretion. It was observed that higher than 10 μg/mL (20 μM) cyanidin-3-O-glucoside concentrations did not significantly improve the insulin secretion in rodent pancreatic β-cells INS-1 832/13 [8]. Some in vivo studies reported that cyanidin-3-O-glucoside and anthocyanin-rich extracts ameliorated the hyperglycemia and the insulin sensitivity in diabetic mice [42, 43]. Insulin gene expression was decreased by exposure to oxidative stress in HIT-T15 cells or isolated rat islets [41].

Bottom Line: The results showed that physiologically achievable concentrations of CAE (1, 5, and 10 μM) protect βTC3 against H2O2- and HG-induced cytotoxicity.Antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were increased in pancreatic β-cells pretreated with CAE compared to cells exposed to the prooxidant agents.These results demonstrate that anthocyanins from CAE were biologically active, showing a secretagogue potential and an antioxidative protection of enzymatic systems, conferring protection against H2O2 and glucose toxicity in βTC3 cells.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine, Mănăştur 3-5, 400372 Cluj-Napoca, Romania.

ABSTRACT
The aim of this study was to investigate the protective effects of a chokeberry anthocyanin extract (CAE) on pancreatic β-cells (βTC3) exposed to hydrogen peroxide- (H2O2-) and high glucose- (HG-) induced oxidative stress conditions. In order to quantify individual anthocyanins high performance liquid chromatography (HPLC) coupled to photodiode array (PDA) was used. The identification of the fragment ion pattern of anthocyanins was carried out by electrospray ionization mass spectrometry (LC-ESI-MS). The results showed that physiologically achievable concentrations of CAE (1, 5, and 10 μM) protect βTC3 against H2O2- and HG-induced cytotoxicity. Antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were increased in pancreatic β-cells pretreated with CAE compared to cells exposed to the prooxidant agents. GSH levels initially reduced after exposure to H2O2 and HG were restored by pretreatment with CAE. Insulin secretion in βTC3 cells was enhanced by CAE pretreatment. CAE restored the insulin pool and diminished the intracellular reactive oxygen species level in glucose-induced stress condition in βTC3 cells. These results demonstrate that anthocyanins from CAE were biologically active, showing a secretagogue potential and an antioxidative protection of enzymatic systems, conferring protection against H2O2 and glucose toxicity in βTC3 cells.

No MeSH data available.


Related in: MedlinePlus