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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 the cell cycle of IMR90 cells. IMR90, normal human cells, were synchronized by contact inhibition and used to seed new cultures. One group of the new cultures was assigned to 100 nM MB while the other group remained as control. Both groups were incubated for 18, 24, 48, and 72 h. At the end of each time point the cells were harvested, washed with ice-cold PBS, and prepared for flow cytometry analysis for the various stages of the cell cycle as described in Material and Methods. The data from the flow cytometer was used for determining the distribution of the various stages of cell cycle with the help of ModFit software (Verity Software, Topsham, ME). A. Shows the synchronization of the cells and the distribution of the cell cycle stages in 90% confluent culture and in over confluent culture. Each time point represents the Mean±sem of three independent experiments, each performed in triplicates. *P<0.05 One-way ANOVA, Friedman test. B. Shows the distribution of the cell cycle stages in MB-treated and control cultures at the time point 48 h. Although several intervals were tested, shown are the data from 48 h incubation with MB (see text for details).
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f0045: The effect of MB on the cell cycle of IMR90 cells. IMR90, normal human cells, were synchronized by contact inhibition and used to seed new cultures. One group of the new cultures was assigned to 100 nM MB while the other group remained as control. Both groups were incubated for 18, 24, 48, and 72 h. At the end of each time point the cells were harvested, washed with ice-cold PBS, and prepared for flow cytometry analysis for the various stages of the cell cycle as described in Material and Methods. The data from the flow cytometer was used for determining the distribution of the various stages of cell cycle with the help of ModFit software (Verity Software, Topsham, ME). A. Shows the synchronization of the cells and the distribution of the cell cycle stages in 90% confluent culture and in over confluent culture. Each time point represents the Mean±sem of three independent experiments, each performed in triplicates. *P<0.05 One-way ANOVA, Friedman test. B. Shows the distribution of the cell cycle stages in MB-treated and control cultures at the time point 48 h. Although several intervals were tested, shown are the data from 48 h incubation with MB (see text for details).

Mentions: Since IMR90 are normal cells, the effect of MB on the cell cycle was investigated by measuring the distribution of the different stages of the cell cycle in synchronized IMR90 cell culture. Synchronization by contact inhibition caused more than 95% of the cells to enter G1 phase (Fig. 9A). The effect of MB on the cell cycle of the synchronized cultures was examined at 18, 24, 48, and 72 h. MB did not alter the cell cycle stages regardless of the duration of exposure to MB. The data presented in Fig. 9B was selected from IMR90 cells that were exposed to MB for 48 h (Fig. 9B).


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 the cell cycle of IMR90 cells. IMR90, normal human cells, were synchronized by contact inhibition and used to seed new cultures. One group of the new cultures was assigned to 100 nM MB while the other group remained as control. Both groups were incubated for 18, 24, 48, and 72 h. At the end of each time point the cells were harvested, washed with ice-cold PBS, and prepared for flow cytometry analysis for the various stages of the cell cycle as described in Material and Methods. The data from the flow cytometer was used for determining the distribution of the various stages of cell cycle with the help of ModFit software (Verity Software, Topsham, ME). A. Shows the synchronization of the cells and the distribution of the cell cycle stages in 90% confluent culture and in over confluent culture. Each time point represents the Mean±sem of three independent experiments, each performed in triplicates. *P<0.05 One-way ANOVA, Friedman test. B. Shows the distribution of the cell cycle stages in MB-treated and control cultures at the time point 48 h. Although several intervals were tested, shown are the data from 48 h incubation with MB (see text for details).
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

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

f0045: The effect of MB on the cell cycle of IMR90 cells. IMR90, normal human cells, were synchronized by contact inhibition and used to seed new cultures. One group of the new cultures was assigned to 100 nM MB while the other group remained as control. Both groups were incubated for 18, 24, 48, and 72 h. At the end of each time point the cells were harvested, washed with ice-cold PBS, and prepared for flow cytometry analysis for the various stages of the cell cycle as described in Material and Methods. The data from the flow cytometer was used for determining the distribution of the various stages of cell cycle with the help of ModFit software (Verity Software, Topsham, ME). A. Shows the synchronization of the cells and the distribution of the cell cycle stages in 90% confluent culture and in over confluent culture. Each time point represents the Mean±sem of three independent experiments, each performed in triplicates. *P<0.05 One-way ANOVA, Friedman test. B. Shows the distribution of the cell cycle stages in MB-treated and control cultures at the time point 48 h. Although several intervals were tested, shown are the data from 48 h incubation with MB (see text for details).
Mentions: Since IMR90 are normal cells, the effect of MB on the cell cycle was investigated by measuring the distribution of the different stages of the cell cycle in synchronized IMR90 cell culture. Synchronization by contact inhibition caused more than 95% of the cells to enter G1 phase (Fig. 9A). The effect of MB on the cell cycle of the synchronized cultures was examined at 18, 24, 48, and 72 h. MB did not alter the cell cycle stages regardless of the duration of exposure to MB. The data presented in Fig. 9B was selected from IMR90 cells that were exposed to MB for 48 h (Fig. 9B).

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