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Combined activation of the energy and cellular-defense pathways may explain the potent anti-senescence activity of methylene blue.

Atamna H, Atamna W, Al-Eyd G, Shanower G, Dhahbi JM - Redox Biol (2015)

Bottom Line: Methylene blue (MB) delays cellular senescence, induces complex-IV, and activates Keap1/Nrf2; however, the molecular link of these effects to MB is unclear.A previous research suggested that the pattern of AMPK activation (i.e., chronic or transient) determines the AMPK effect on cell senescence.Since MB lacked an effect on cell cycle, an MB-dependent change to cell cycle is unlikely to contribute to the anti-senescence activity.

View Article: PubMed Central - PubMed

Affiliation: College of Medicine, California University of Science & Medicine, Colton, CA 92324, USA; Department of Basic Sciences, The Commonwealth Medical College (TCMC), Scranton, PA 18509, USA. Electronic address: atamnah@calmedu.org.

No MeSH data available.


Related in: MedlinePlus

The effect of MB on intracellular oxidants. IMR90 cell cultures were treated with 100 nM MB for increasing intervals (up to 7 days). The cells were harvested by trypsinazation, counted, and resuspended into 25 mM HEPES (pH 7.3)/DMEM. One million cells of each specific time point were placed in the designated tube and incubated in dark with 1 µM of 2′,7′-dichlorfluorescein-diacetate (DCFH-DA) and the fluorescence of DCF was measured using flow cytometry as described in Section 2. The data shown are the percentages of the Mean AFU±sem of six independent experiments, each performed in triplicates. *P<0.01, (One-way ANOVA, Dunnett's multiple comparisons test).
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f0035: The effect of MB on intracellular oxidants. IMR90 cell cultures were treated with 100 nM MB for increasing intervals (up to 7 days). The cells were harvested by trypsinazation, counted, and resuspended into 25 mM HEPES (pH 7.3)/DMEM. One million cells of each specific time point were placed in the designated tube and incubated in dark with 1 µM of 2′,7′-dichlorfluorescein-diacetate (DCFH-DA) and the fluorescence of DCF was measured using flow cytometry as described in Section 2. The data shown are the percentages of the Mean AFU±sem of six independent experiments, each performed in triplicates. *P<0.01, (One-way ANOVA, Dunnett's multiple comparisons test).

Mentions: Elevated activity of complex IV improves mitochondrial respiration [23] and lowers cellular level of oxidants [41–43]. Therefore we measured the level of oxidants in MB-treated cells. The steady-state level oxidants level in MB-treated cells was 28% lower than that of the control (Fig. 7). Intracellular oxidants were determined by using DCFH-DA.


Combined activation of the energy and cellular-defense pathways may explain the potent anti-senescence activity of methylene blue.

Atamna H, Atamna W, Al-Eyd G, Shanower G, Dhahbi JM - Redox Biol (2015)

The effect of MB on intracellular oxidants. IMR90 cell cultures were treated with 100 nM MB for increasing intervals (up to 7 days). The cells were harvested by trypsinazation, counted, and resuspended into 25 mM HEPES (pH 7.3)/DMEM. One million cells of each specific time point were placed in the designated tube and incubated in dark with 1 µM of 2′,7′-dichlorfluorescein-diacetate (DCFH-DA) and the fluorescence of DCF was measured using flow cytometry as described in Section 2. The data shown are the percentages of the Mean AFU±sem of six independent experiments, each performed in triplicates. *P<0.01, (One-way ANOVA, Dunnett's multiple comparisons test).
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4588422&req=5

f0035: The effect of MB on intracellular oxidants. IMR90 cell cultures were treated with 100 nM MB for increasing intervals (up to 7 days). The cells were harvested by trypsinazation, counted, and resuspended into 25 mM HEPES (pH 7.3)/DMEM. One million cells of each specific time point were placed in the designated tube and incubated in dark with 1 µM of 2′,7′-dichlorfluorescein-diacetate (DCFH-DA) and the fluorescence of DCF was measured using flow cytometry as described in Section 2. The data shown are the percentages of the Mean AFU±sem of six independent experiments, each performed in triplicates. *P<0.01, (One-way ANOVA, Dunnett's multiple comparisons test).
Mentions: Elevated activity of complex IV improves mitochondrial respiration [23] and lowers cellular level of oxidants [41–43]. Therefore we measured the level of oxidants in MB-treated cells. The steady-state level oxidants level in MB-treated cells was 28% lower than that of the control (Fig. 7). Intracellular oxidants were determined by using DCFH-DA.

Bottom Line: Methylene blue (MB) delays cellular senescence, induces complex-IV, and activates Keap1/Nrf2; however, the molecular link of these effects to MB is unclear.A previous research suggested that the pattern of AMPK activation (i.e., chronic or transient) determines the AMPK effect on cell senescence.Since MB lacked an effect on cell cycle, an MB-dependent change to cell cycle is unlikely to contribute to the anti-senescence activity.

View Article: PubMed Central - PubMed

Affiliation: College of Medicine, California University of Science & Medicine, Colton, CA 92324, USA; Department of Basic Sciences, The Commonwealth Medical College (TCMC), Scranton, PA 18509, USA. Electronic address: atamnah@calmedu.org.

No MeSH data available.


Related in: MedlinePlus