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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 the levels of antioxidant enzyme activities in βTC3 cells in stress-induced conditions. (a) HG-induced stress for 24 h: (A) SOD; (B) CAT; (C) GPx; (D) GSH and (b) H2O2-induced stress for 15 min: (A) SOD; (B) CAT; (C) GPx; (D) GSH. Data are expressed as mean ± SEM of two independent determinations. Statistical analyses: ∗P < 0.05 versus the control.
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fig4: Effect of CAE on the levels of antioxidant enzyme activities in βTC3 cells in stress-induced conditions. (a) HG-induced stress for 24 h: (A) SOD; (B) CAT; (C) GPx; (D) GSH and (b) H2O2-induced stress for 15 min: (A) SOD; (B) CAT; (C) GPx; (D) GSH. Data are expressed as mean ± SEM of two independent determinations. Statistical analyses: ∗P < 0.05 versus the control.

Mentions: In the presence of H2O2 and HG as stress agents, the CAE induced a strong increase of CAT, GPx activities, and GSH content in β-pancreatic TC3 cells, as shown in Figures 4(a) and 4(b). Also, a slight increase in the activity of SOD was observed in CAE-treated β-pancreatic cells compared to untreated cells; see Figures 4(a)(A) and 4(b)(A). Since CAT and GPx have different enzymatic characteristic, both of them are H2O2-inactivating enzymes. GPx inactivates the H2O2 substrate with high affinity in a continuous rate of oxygen free radical production, whereas CAT has a lower affinity in a higher continuous rate of free ROS production [35]. A possible explanation for the statistically significant increased activities of CAT and GPx by CAE treatment in oxidative stress-induced conditions (50 mM HG and 70 μM H2O2 in β-pancreatic TC3 cells) could be the high rate production of their substrate H2O2 and a scarce existence of superoxide anion (O2•−) (Figures 4(a)(B) and 4(a)(C)).


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 the levels of antioxidant enzyme activities in βTC3 cells in stress-induced conditions. (a) HG-induced stress for 24 h: (A) SOD; (B) CAT; (C) GPx; (D) GSH and (b) H2O2-induced stress for 15 min: (A) SOD; (B) CAT; (C) GPx; (D) GSH. Data are expressed as mean ± SEM of two independent determinations. Statistical analyses: ∗P < 0.05 versus the control.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4465716&req=5

fig4: Effect of CAE on the levels of antioxidant enzyme activities in βTC3 cells in stress-induced conditions. (a) HG-induced stress for 24 h: (A) SOD; (B) CAT; (C) GPx; (D) GSH and (b) H2O2-induced stress for 15 min: (A) SOD; (B) CAT; (C) GPx; (D) GSH. Data are expressed as mean ± SEM of two independent determinations. Statistical analyses: ∗P < 0.05 versus the control.
Mentions: In the presence of H2O2 and HG as stress agents, the CAE induced a strong increase of CAT, GPx activities, and GSH content in β-pancreatic TC3 cells, as shown in Figures 4(a) and 4(b). Also, a slight increase in the activity of SOD was observed in CAE-treated β-pancreatic cells compared to untreated cells; see Figures 4(a)(A) and 4(b)(A). Since CAT and GPx have different enzymatic characteristic, both of them are H2O2-inactivating enzymes. GPx inactivates the H2O2 substrate with high affinity in a continuous rate of oxygen free radical production, whereas CAT has a lower affinity in a higher continuous rate of free ROS production [35]. A possible explanation for the statistically significant increased activities of CAT and GPx by CAE treatment in oxidative stress-induced conditions (50 mM HG and 70 μM H2O2 in β-pancreatic TC3 cells) could be the high rate production of their substrate H2O2 and a scarce existence of superoxide anion (O2•−) (Figures 4(a)(B) and 4(a)(C)).

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