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Polyphenolic composition of grape stem extracts affects antioxidant activity in endothelial and muscle cells.

Goutzourelas N, Stagos D, Spanidis Y, Liosi M, Apostolou A, Priftis A, Haroutounian S, Spandidos DA, Tsatsakis AM, Kouretas D - Mol Med Rep (2015)

Bottom Line: The results revealed that treatment of the EA.hy926 cells with Mandilaria extract significantly decreased the TBARS levels by 14.8% and the CARB levels by 25.9 %, while it increased the GSH levels by 15.8% compared to the controls.Treatment of the C2C12 cells with Mandilaria extract significantly decreased the TBARS levels by 47.3 %, the CARB levels by 39.0 % and the ROS levels by 21.8%, while it increased the GSH levels by 22.6% compared to the controls.In conclusion, to the best of our knowledgel, our results demonstrate for the first time that treatment with grape stem extracts at low concentrations improves the redox status of endothelial and muscle cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Biotechnology, University of Thessaly, 41221 Larissa, Greece.

ABSTRACT
The aim of the present study was the assessment of the antioxidant effects of polyphenolic extracts derived from the stems of three Greek grape varieties (Moshomayro, Mavrotragano and Mandilaria) in endothelial (EA.hy926) and muscle (C2C12) cells. We also investigated the effects of the polyphenolic composition on the antioxidant effects of the grape stem extracts. For this purpose, the endothelial and muscle cells were treated with low non-cytotoxic concentrations of the extracts for 24 h in order to assess the effects of the extracts on cellular redox status using oxidative stress biomarkers. The oxidative stress markers were thiobarbituric acid reactive substances (TBARS), protein carbonyl (CARB) levels, reactive oxygen species (ROS) levels and glutathione (GSH) levels. The results revealed that treatment of the EA.hy926 cells with Mandilaria extract significantly decreased the TBARS levels by 14.8% and the CARB levels by 25.9 %, while it increased the GSH levels by 15.8% compared to the controls. Moreover, treatment of the EA.hy926 cells with Mavrotragano extract significantly increased the GSH levels by 20.2%, while it significantly decreased the TBARS and CARB levels by 12.5% and 16.6%, respectively. Treatment of the C2C12 cells with Mandilaria extract significantly decreased the TBARS levels by 47.3 %, the CARB levels by 39.0 % and the ROS levels by 21.8%, while it increased the GSH levels by 22.6% compared to the controls. Moreover, treatment of the C2C12 cells with Mavrotragano significantly decreased the TBARS, CARB and ROS levels by 36.2%, 35.9% and 16.5%, respectively. In conclusion, to the best of our knowledgel, our results demonstrate for the first time that treatment with grape stem extracts at low concentrations improves the redox status of endothelial and muscle cells. Thus, grape stem extracts may be used for developing antioxidant food supplements or biofunctional foods. However, it was also found that the polyphenolic composition of grape stem extracts affects their antioxidant capacity. For example, the results suggested that trans-resveratrol, gallic acid, (+)-catechin, ferulic acid, caffeic acid, quercetin, coumaric acid and kaempferol may be essential for the antioxidant activity of grape stem extracts.

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Effects of treatment with grape stem extracts for 24 h on levels of oxidative stress markers in EA.hy926 cells. (A) The histogram of cell counts versus fluorescence of 10,000 cells analyzed by the flow cytometry for the detection of glutathione (GSH) levels. FL2 represents the detection of fluorescence using 488 and 580 nm as the excitation and emission wavelength, respectively. (B) The histograms show the cell counts versus fluorescence of 10,000 cells analyzed by the flow cytometry for the detection of reactive oxygen species (ROS) levels. FL1 represents the detection of fluorescence using 488 and 530 nm as the excitation and emission wavelength, respectively. (C) Effects on ROS, GSH, thiobarbituric acid reactive substances (TBARS) and protein carbonyl (CARB) levels. TBARS and CARB levels were assessed using a spectrophotometer. All values are presented as the means ± SEM of three independent experiments. *p<0.05 indicates significant difference compared to the controls (untreated cells).
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f3-mmr-12-04-5846: Effects of treatment with grape stem extracts for 24 h on levels of oxidative stress markers in EA.hy926 cells. (A) The histogram of cell counts versus fluorescence of 10,000 cells analyzed by the flow cytometry for the detection of glutathione (GSH) levels. FL2 represents the detection of fluorescence using 488 and 580 nm as the excitation and emission wavelength, respectively. (B) The histograms show the cell counts versus fluorescence of 10,000 cells analyzed by the flow cytometry for the detection of reactive oxygen species (ROS) levels. FL1 represents the detection of fluorescence using 488 and 530 nm as the excitation and emission wavelength, respectively. (C) Effects on ROS, GSH, thiobarbituric acid reactive substances (TBARS) and protein carbonyl (CARB) levels. TBARS and CARB levels were assessed using a spectrophotometer. All values are presented as the means ± SEM of three independent experiments. *p<0.05 indicates significant difference compared to the controls (untreated cells).

Mentions: The levels of oxidative stress markers were measured in order to assess effects of the extracts on the redox status of EA.hy926cells. The ROS and GSH levels were measured by flow cytometry using DCF-DA and mercury orange, respectively. Histograms demonstrating the cell counts versus the fluorescence of the mercury orange or DCF-DA are shown in Fig. 3A and B, respectively. The mean fluorescence intensity values were evaluated using BD CellQuest software and are expressed as a percentage of the controls (untreated cells). None of the tested extracts had any effect on the ROS levels (Fig. 3C). Moreover, the Mavrotragano and Mandilaria extracts significantly increased the GSH levels by 20.2±1.5 and 15.8±4.1%, respectively compared to the controls (Fig. 3C). On the contrary, Moshomavro extract did not affect the GSH levels.


Polyphenolic composition of grape stem extracts affects antioxidant activity in endothelial and muscle cells.

Goutzourelas N, Stagos D, Spanidis Y, Liosi M, Apostolou A, Priftis A, Haroutounian S, Spandidos DA, Tsatsakis AM, Kouretas D - Mol Med Rep (2015)

Effects of treatment with grape stem extracts for 24 h on levels of oxidative stress markers in EA.hy926 cells. (A) The histogram of cell counts versus fluorescence of 10,000 cells analyzed by the flow cytometry for the detection of glutathione (GSH) levels. FL2 represents the detection of fluorescence using 488 and 580 nm as the excitation and emission wavelength, respectively. (B) The histograms show the cell counts versus fluorescence of 10,000 cells analyzed by the flow cytometry for the detection of reactive oxygen species (ROS) levels. FL1 represents the detection of fluorescence using 488 and 530 nm as the excitation and emission wavelength, respectively. (C) Effects on ROS, GSH, thiobarbituric acid reactive substances (TBARS) and protein carbonyl (CARB) levels. TBARS and CARB levels were assessed using a spectrophotometer. All values are presented as the means ± SEM of three independent experiments. *p<0.05 indicates significant difference compared to the controls (untreated cells).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581749&req=5

f3-mmr-12-04-5846: Effects of treatment with grape stem extracts for 24 h on levels of oxidative stress markers in EA.hy926 cells. (A) The histogram of cell counts versus fluorescence of 10,000 cells analyzed by the flow cytometry for the detection of glutathione (GSH) levels. FL2 represents the detection of fluorescence using 488 and 580 nm as the excitation and emission wavelength, respectively. (B) The histograms show the cell counts versus fluorescence of 10,000 cells analyzed by the flow cytometry for the detection of reactive oxygen species (ROS) levels. FL1 represents the detection of fluorescence using 488 and 530 nm as the excitation and emission wavelength, respectively. (C) Effects on ROS, GSH, thiobarbituric acid reactive substances (TBARS) and protein carbonyl (CARB) levels. TBARS and CARB levels were assessed using a spectrophotometer. All values are presented as the means ± SEM of three independent experiments. *p<0.05 indicates significant difference compared to the controls (untreated cells).
Mentions: The levels of oxidative stress markers were measured in order to assess effects of the extracts on the redox status of EA.hy926cells. The ROS and GSH levels were measured by flow cytometry using DCF-DA and mercury orange, respectively. Histograms demonstrating the cell counts versus the fluorescence of the mercury orange or DCF-DA are shown in Fig. 3A and B, respectively. The mean fluorescence intensity values were evaluated using BD CellQuest software and are expressed as a percentage of the controls (untreated cells). None of the tested extracts had any effect on the ROS levels (Fig. 3C). Moreover, the Mavrotragano and Mandilaria extracts significantly increased the GSH levels by 20.2±1.5 and 15.8±4.1%, respectively compared to the controls (Fig. 3C). On the contrary, Moshomavro extract did not affect the GSH levels.

Bottom Line: The results revealed that treatment of the EA.hy926 cells with Mandilaria extract significantly decreased the TBARS levels by 14.8% and the CARB levels by 25.9 %, while it increased the GSH levels by 15.8% compared to the controls.Treatment of the C2C12 cells with Mandilaria extract significantly decreased the TBARS levels by 47.3 %, the CARB levels by 39.0 % and the ROS levels by 21.8%, while it increased the GSH levels by 22.6% compared to the controls.In conclusion, to the best of our knowledgel, our results demonstrate for the first time that treatment with grape stem extracts at low concentrations improves the redox status of endothelial and muscle cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Biotechnology, University of Thessaly, 41221 Larissa, Greece.

ABSTRACT
The aim of the present study was the assessment of the antioxidant effects of polyphenolic extracts derived from the stems of three Greek grape varieties (Moshomayro, Mavrotragano and Mandilaria) in endothelial (EA.hy926) and muscle (C2C12) cells. We also investigated the effects of the polyphenolic composition on the antioxidant effects of the grape stem extracts. For this purpose, the endothelial and muscle cells were treated with low non-cytotoxic concentrations of the extracts for 24 h in order to assess the effects of the extracts on cellular redox status using oxidative stress biomarkers. The oxidative stress markers were thiobarbituric acid reactive substances (TBARS), protein carbonyl (CARB) levels, reactive oxygen species (ROS) levels and glutathione (GSH) levels. The results revealed that treatment of the EA.hy926 cells with Mandilaria extract significantly decreased the TBARS levels by 14.8% and the CARB levels by 25.9 %, while it increased the GSH levels by 15.8% compared to the controls. Moreover, treatment of the EA.hy926 cells with Mavrotragano extract significantly increased the GSH levels by 20.2%, while it significantly decreased the TBARS and CARB levels by 12.5% and 16.6%, respectively. Treatment of the C2C12 cells with Mandilaria extract significantly decreased the TBARS levels by 47.3 %, the CARB levels by 39.0 % and the ROS levels by 21.8%, while it increased the GSH levels by 22.6% compared to the controls. Moreover, treatment of the C2C12 cells with Mavrotragano significantly decreased the TBARS, CARB and ROS levels by 36.2%, 35.9% and 16.5%, respectively. In conclusion, to the best of our knowledgel, our results demonstrate for the first time that treatment with grape stem extracts at low concentrations improves the redox status of endothelial and muscle cells. Thus, grape stem extracts may be used for developing antioxidant food supplements or biofunctional foods. However, it was also found that the polyphenolic composition of grape stem extracts affects their antioxidant capacity. For example, the results suggested that trans-resveratrol, gallic acid, (+)-catechin, ferulic acid, caffeic acid, quercetin, coumaric acid and kaempferol may be essential for the antioxidant activity of grape stem extracts.

Show MeSH
Related in: MedlinePlus