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Preconditioning L6 Muscle Cells with Naringin Ameliorates Oxidative Stress and Increases Glucose Uptake.

Dhanya R, Arun KB, Nisha VM, Syama HP, Nisha P, Santhosh Kumar TR, Jayamurthy P - PLoS ONE (2015)

Bottom Line: Thus, strategies to reduce oxidative stress may alleviate these pathogenic processes.Addition of Naringin (100 μM) showed approximately 40% reduction in protein glycation in vitro.Our results demonstrate that Naringin activate GSH synthesis through a novel antioxidant defense mechanism against excessive Reactive Oxygen Species (ROS) production, contributing to the prevention of oxidative damage in addition to its effect on glycemic control.

View Article: PubMed Central - PubMed

Affiliation: Agroprocessing and Natural Products Division, National Institute for Interdisciplinary Science and Technology (NIIST), CSIR, Industrial Estate, Pappanamcode, Thiruvananthapuram-695019, Kerala, India.

ABSTRACT
Enhanced oxidative stress contributes to pathological changes in diabetes and its complications. Thus, strategies to reduce oxidative stress may alleviate these pathogenic processes. Herein, we have investigated Naringin mediated regulation of glutathione (GSH) & intracellular free radical levels and modulation of glucose uptake under oxidative stress in L6 cell lines. The results from the study demonstrated a marked decrease in glutathione with a subsequent increase in free radical levels, which was reversed by the pretreatment of Naringin. We also observed that the increased malondialdehyde level, the marker of lipid peroxidation on induction of oxidative stress was retrieved on Naringin pretreatment. Addition of Naringin (100 μM) showed approximately 40% reduction in protein glycation in vitro. Furthermore, we observed a twofold increase in uptake of fluorescent labeled glucose namely 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose (2-NBDG) on Naringin treatment in differentiated L6 myoblast. The increased uptake of 2-NBDG by L6 myotubes may be attributed due to the enhanced translocation of GLUT4. Our results demonstrate that Naringin activate GSH synthesis through a novel antioxidant defense mechanism against excessive Reactive Oxygen Species (ROS) production, contributing to the prevention of oxidative damage in addition to its effect on glycemic control.

No MeSH data available.


Related in: MedlinePlus

Fluorescence analysis of 2-NBDG uptake by flow cytometry.FACS analysis of 2-NBDG uptake in differentiated L6 cells by plotting cell count against FITC revealed that 8%, 8.1% and 30% of cells uptake 2-NBDG in control, TBHP and Rosiglitazone treated cells respectively whereas 30.6%, 33.1%, 28%, 32% of cells uptake 2-NBDG, pretreated with two different concentrations (10 and 100 μM) of Naringin along with/without TBHP respectively. Each value represents mean ± SD (standard deviation) from triplicate measurements (n = 3) of three different experiments. Significance test between various groups was determined by using one way ANOVA followed by Duncan’s multiple range test.* P≤0.05 versus control.
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pone.0132429.g004: Fluorescence analysis of 2-NBDG uptake by flow cytometry.FACS analysis of 2-NBDG uptake in differentiated L6 cells by plotting cell count against FITC revealed that 8%, 8.1% and 30% of cells uptake 2-NBDG in control, TBHP and Rosiglitazone treated cells respectively whereas 30.6%, 33.1%, 28%, 32% of cells uptake 2-NBDG, pretreated with two different concentrations (10 and 100 μM) of Naringin along with/without TBHP respectively. Each value represents mean ± SD (standard deviation) from triplicate measurements (n = 3) of three different experiments. Significance test between various groups was determined by using one way ANOVA followed by Duncan’s multiple range test.* P≤0.05 versus control.

Mentions: 2-NBDG uptake in L6 myotubes was analyzed by flow cytometry by detecting the fluorescence within the cells. There was no significant effect on 2-NBDG uptake in the cell induced with TBHP (8.7%) compared to control (8%) (Fig 4). At 24 h pretreatment of Naringin (10 μM and 100 μm), the glucose uptake in L6 myotubes remarkably increased to 27.5% and 35.9%, which was much higher than that of positive control, Rosiglitazone (30.4%, P≥0.05).


Preconditioning L6 Muscle Cells with Naringin Ameliorates Oxidative Stress and Increases Glucose Uptake.

Dhanya R, Arun KB, Nisha VM, Syama HP, Nisha P, Santhosh Kumar TR, Jayamurthy P - PLoS ONE (2015)

Fluorescence analysis of 2-NBDG uptake by flow cytometry.FACS analysis of 2-NBDG uptake in differentiated L6 cells by plotting cell count against FITC revealed that 8%, 8.1% and 30% of cells uptake 2-NBDG in control, TBHP and Rosiglitazone treated cells respectively whereas 30.6%, 33.1%, 28%, 32% of cells uptake 2-NBDG, pretreated with two different concentrations (10 and 100 μM) of Naringin along with/without TBHP respectively. Each value represents mean ± SD (standard deviation) from triplicate measurements (n = 3) of three different experiments. Significance test between various groups was determined by using one way ANOVA followed by Duncan’s multiple range test.* P≤0.05 versus control.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4492986&req=5

pone.0132429.g004: Fluorescence analysis of 2-NBDG uptake by flow cytometry.FACS analysis of 2-NBDG uptake in differentiated L6 cells by plotting cell count against FITC revealed that 8%, 8.1% and 30% of cells uptake 2-NBDG in control, TBHP and Rosiglitazone treated cells respectively whereas 30.6%, 33.1%, 28%, 32% of cells uptake 2-NBDG, pretreated with two different concentrations (10 and 100 μM) of Naringin along with/without TBHP respectively. Each value represents mean ± SD (standard deviation) from triplicate measurements (n = 3) of three different experiments. Significance test between various groups was determined by using one way ANOVA followed by Duncan’s multiple range test.* P≤0.05 versus control.
Mentions: 2-NBDG uptake in L6 myotubes was analyzed by flow cytometry by detecting the fluorescence within the cells. There was no significant effect on 2-NBDG uptake in the cell induced with TBHP (8.7%) compared to control (8%) (Fig 4). At 24 h pretreatment of Naringin (10 μM and 100 μm), the glucose uptake in L6 myotubes remarkably increased to 27.5% and 35.9%, which was much higher than that of positive control, Rosiglitazone (30.4%, P≥0.05).

Bottom Line: Thus, strategies to reduce oxidative stress may alleviate these pathogenic processes.Addition of Naringin (100 μM) showed approximately 40% reduction in protein glycation in vitro.Our results demonstrate that Naringin activate GSH synthesis through a novel antioxidant defense mechanism against excessive Reactive Oxygen Species (ROS) production, contributing to the prevention of oxidative damage in addition to its effect on glycemic control.

View Article: PubMed Central - PubMed

Affiliation: Agroprocessing and Natural Products Division, National Institute for Interdisciplinary Science and Technology (NIIST), CSIR, Industrial Estate, Pappanamcode, Thiruvananthapuram-695019, Kerala, India.

ABSTRACT
Enhanced oxidative stress contributes to pathological changes in diabetes and its complications. Thus, strategies to reduce oxidative stress may alleviate these pathogenic processes. Herein, we have investigated Naringin mediated regulation of glutathione (GSH) & intracellular free radical levels and modulation of glucose uptake under oxidative stress in L6 cell lines. The results from the study demonstrated a marked decrease in glutathione with a subsequent increase in free radical levels, which was reversed by the pretreatment of Naringin. We also observed that the increased malondialdehyde level, the marker of lipid peroxidation on induction of oxidative stress was retrieved on Naringin pretreatment. Addition of Naringin (100 μM) showed approximately 40% reduction in protein glycation in vitro. Furthermore, we observed a twofold increase in uptake of fluorescent labeled glucose namely 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose (2-NBDG) on Naringin treatment in differentiated L6 myoblast. The increased uptake of 2-NBDG by L6 myotubes may be attributed due to the enhanced translocation of GLUT4. Our results demonstrate that Naringin activate GSH synthesis through a novel antioxidant defense mechanism against excessive Reactive Oxygen Species (ROS) production, contributing to the prevention of oxidative damage in addition to its effect on glycemic control.

No MeSH data available.


Related in: MedlinePlus