Limits...
Wild Raspberry Subjected to Simulated Gastrointestinal Digestion Improves the Protective Capacity against Ethyl Carbamate-Induced Oxidative Damage in Caco-2 Cells.

Chen W, Xu Y, Zhang L, Li Y, Zheng X - Oxid Med Cell Longev (2015)

Bottom Line: Wild raspberries are rich in polyphenolic compounds, which possess potent antioxidant activity.In addition, HPLC-ESI-MS results showed that the contents of identified polyphenolic compounds (esculin, kaempferol O-hexoside, and pelargonidin O-hexoside) were remarkably increased after digestion, which might be related to the better protective effect of RD.Overall, our results demonstrated that raspberry extract undergoing simulated gastrointestinal digestion may improve the protective effect against EC-induced oxidative damage in Caco-2 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Food Science and Nutrition, Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang University, Hangzhou 310058, China.

ABSTRACT
Ethyl carbamate (EC), a probable human carcinogen, occurs widely in many fermented foods. Previous studies indicated that EC-induced cytotoxicity was associated with oxidative stress. Wild raspberries are rich in polyphenolic compounds, which possess potent antioxidant activity. This study was conducted to investigate the protective effect of wild raspberry extracts produced before (RE) and after in vitro simulated gastrointestinal digestion (RD) on EC-induced oxidative damage in Caco-2 cells. Our primary data showed that ethyl carbamate could result in cytotoxicity and genotoxicity in Caco-2 cells and raspberry extract after digestion (RD) may be more effective than that before digestion (RE) in attenuating toxicity caused by ethyl carbamate. Further investigation by fluorescence microscope revealed that RD may significantly ameliorate EC-induced oxidative damage by scavenging the overproduction of intracellular reactive oxygen species (ROS), maintaining mitochondrial function and preventing glutathione (GSH) depletion. In addition, HPLC-ESI-MS results showed that the contents of identified polyphenolic compounds (esculin, kaempferol O-hexoside, and pelargonidin O-hexoside) were remarkably increased after digestion, which might be related to the better protective effect of RD. Overall, our results demonstrated that raspberry extract undergoing simulated gastrointestinal digestion may improve the protective effect against EC-induced oxidative damage in Caco-2 cells.

No MeSH data available.


Related in: MedlinePlus

Effect of RD on EC-induced oxidative damage to mitochondrial membrane in Caco-2 cells. (a) After treatment with 62.5 mM EC in the presence or absence of RE (2 mg/mL) or RD (2 mg/mL) for 24 h, Caco-2 cells were incubated with 10 μM NAO for 30 min and subsequently adopted to fluorescence microscope analysis. (b) The quantitative data of panel (a) and results were expressed as mean NAO fluorescence intensity (mean ± standard deviations). ∗p < 0.05 represents significant difference compared with EC group.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4696417&req=5

fig6: Effect of RD on EC-induced oxidative damage to mitochondrial membrane in Caco-2 cells. (a) After treatment with 62.5 mM EC in the presence or absence of RE (2 mg/mL) or RD (2 mg/mL) for 24 h, Caco-2 cells were incubated with 10 μM NAO for 30 min and subsequently adopted to fluorescence microscope analysis. (b) The quantitative data of panel (a) and results were expressed as mean NAO fluorescence intensity (mean ± standard deviations). ∗p < 0.05 represents significant difference compared with EC group.

Mentions: In addition, it is reported that overproduction of ROS can result in mitochondrial membrane lipid peroxidation [28]. Thus, we further studied the effect of RD on suppressing EC-induced lipid peroxidation in Caco-2 cells. NAO, a fluorescence probe, was designed to detect cardiolipin which is a mitochondrial membrane lipid component and would be oxidized in the presence of ROS. As shown in Figures 6(a) and 6(b), without RE or RD pretreatment, massive cardiolipins were oxidized in exposure to EC, with the NAO fluorescence intensity declining to 45.46% compared with that of control group. Nevertheless, after treatment with RE or RD for 2 h, the result turned out to be that their fluorescence intensity increased to 55.47% and 84.70%, respectively, compared with that of EC group (45.46%). On the basis of these results, it can be concluded that RD may afford protection against EC-induced oxidative damage to mitochondrial membrane.


Wild Raspberry Subjected to Simulated Gastrointestinal Digestion Improves the Protective Capacity against Ethyl Carbamate-Induced Oxidative Damage in Caco-2 Cells.

Chen W, Xu Y, Zhang L, Li Y, Zheng X - Oxid Med Cell Longev (2015)

Effect of RD on EC-induced oxidative damage to mitochondrial membrane in Caco-2 cells. (a) After treatment with 62.5 mM EC in the presence or absence of RE (2 mg/mL) or RD (2 mg/mL) for 24 h, Caco-2 cells were incubated with 10 μM NAO for 30 min and subsequently adopted to fluorescence microscope analysis. (b) The quantitative data of panel (a) and results were expressed as mean NAO fluorescence intensity (mean ± standard deviations). ∗p < 0.05 represents significant difference compared with EC group.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4696417&req=5

fig6: Effect of RD on EC-induced oxidative damage to mitochondrial membrane in Caco-2 cells. (a) After treatment with 62.5 mM EC in the presence or absence of RE (2 mg/mL) or RD (2 mg/mL) for 24 h, Caco-2 cells were incubated with 10 μM NAO for 30 min and subsequently adopted to fluorescence microscope analysis. (b) The quantitative data of panel (a) and results were expressed as mean NAO fluorescence intensity (mean ± standard deviations). ∗p < 0.05 represents significant difference compared with EC group.
Mentions: In addition, it is reported that overproduction of ROS can result in mitochondrial membrane lipid peroxidation [28]. Thus, we further studied the effect of RD on suppressing EC-induced lipid peroxidation in Caco-2 cells. NAO, a fluorescence probe, was designed to detect cardiolipin which is a mitochondrial membrane lipid component and would be oxidized in the presence of ROS. As shown in Figures 6(a) and 6(b), without RE or RD pretreatment, massive cardiolipins were oxidized in exposure to EC, with the NAO fluorescence intensity declining to 45.46% compared with that of control group. Nevertheless, after treatment with RE or RD for 2 h, the result turned out to be that their fluorescence intensity increased to 55.47% and 84.70%, respectively, compared with that of EC group (45.46%). On the basis of these results, it can be concluded that RD may afford protection against EC-induced oxidative damage to mitochondrial membrane.

Bottom Line: Wild raspberries are rich in polyphenolic compounds, which possess potent antioxidant activity.In addition, HPLC-ESI-MS results showed that the contents of identified polyphenolic compounds (esculin, kaempferol O-hexoside, and pelargonidin O-hexoside) were remarkably increased after digestion, which might be related to the better protective effect of RD.Overall, our results demonstrated that raspberry extract undergoing simulated gastrointestinal digestion may improve the protective effect against EC-induced oxidative damage in Caco-2 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Food Science and Nutrition, Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang University, Hangzhou 310058, China.

ABSTRACT
Ethyl carbamate (EC), a probable human carcinogen, occurs widely in many fermented foods. Previous studies indicated that EC-induced cytotoxicity was associated with oxidative stress. Wild raspberries are rich in polyphenolic compounds, which possess potent antioxidant activity. This study was conducted to investigate the protective effect of wild raspberry extracts produced before (RE) and after in vitro simulated gastrointestinal digestion (RD) on EC-induced oxidative damage in Caco-2 cells. Our primary data showed that ethyl carbamate could result in cytotoxicity and genotoxicity in Caco-2 cells and raspberry extract after digestion (RD) may be more effective than that before digestion (RE) in attenuating toxicity caused by ethyl carbamate. Further investigation by fluorescence microscope revealed that RD may significantly ameliorate EC-induced oxidative damage by scavenging the overproduction of intracellular reactive oxygen species (ROS), maintaining mitochondrial function and preventing glutathione (GSH) depletion. In addition, HPLC-ESI-MS results showed that the contents of identified polyphenolic compounds (esculin, kaempferol O-hexoside, and pelargonidin O-hexoside) were remarkably increased after digestion, which might be related to the better protective effect of RD. Overall, our results demonstrated that raspberry extract undergoing simulated gastrointestinal digestion may improve the protective effect against EC-induced oxidative damage in Caco-2 cells.

No MeSH data available.


Related in: MedlinePlus