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HSP-72 accelerated expression in mononuclear cells induced in vivo by acetyl salicylic acid can be reproduced in vitro when combined with H2O2.

Sandoval-Montiel AA, Zentella-de-Piña M, Ventura-Gallegos JL, Frías-González S, López-Macay A, Zentella-Dehesa A - PLoS ONE (2013)

Bottom Line: Among NSAIDs acetyl salicylic acid remains as a valuable tool because of the variety of benefic prophylactic and therapeutic effects.This effect reaches its maximum 2 h after treatment and disappeared after 5 h.The fact that in vitro acetyl salicylic acid alone did not induce this priming effect implies that in vivo other signals are required.

View Article: PubMed Central - PubMed

Affiliation: Departmento de Medicina Genómica y Toxicología Ambiental, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, México, D.F., México.

ABSTRACT

Background: Among NSAIDs acetyl salicylic acid remains as a valuable tool because of the variety of benefic prophylactic and therapeutic effects. Nevertheless, the molecular bases for these responses have not been complete understood. We explored the effect of acetyl salicylic acid on the heat shock response.

Results: Peripheral blood mononuclear cells from rats challenged with acetyl salicylic acid presented a faster kinetics of expression of HSP-72 messenger RNA and protein in response to in vitro heat shock. This effect reaches its maximum 2 h after treatment and disappeared after 5 h. On isolated peripheral blood mononuclear cells from untreated rats, incubation with acetyl salicylic acid was ineffective to produce priming, but this effect was mimicked when the cells were incubated with the combination of H2O2+ ASA.

Conclusions: Administration of acetyl salicylic acid to rats alters HSP-72 expression mechanism in a way that it becomes more efficient in response to in vitro heat shock. The fact that in vitro acetyl salicylic acid alone did not induce this priming effect implies that in vivo other signals are required. Priming could be reproduces in vitro with the combination of acetyl salicylic acid+H2O2.

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Time course of changes in HSP-72 expression before (T0), immediately after heat shock (HS) and 1 (1 h) and 2 (2 h) hours after recovery at 37°C in peripheral blood mononuclear cells (PBMCs) from untreated rats incubated with or without the combination ASA (10 µM)+H2O2 (5 µM).A) Western blots of HSP-72 from rat PBMCs (100 µg of total protein) incubated with (+) or without (-) the indicated treatments for 1 h before heat shock, β-actin was used as loading control. Protein extracts were obtained before in vitro incubation (T0), or immediately after Heat shock (HS) or (1h) and 2 (2h) hours after recovery at 37°C. B) Histogram of the average signals expressed as Arbitrary units (AU) = OD for HSP-72/OD for β-actin from three independent experiments. *Comparison of HSP-72 content immediately after heat shock between cells without any treatment an those incubated with the combination ASA+H2O2 (n = 3) (p<0.001). **Comparison of cells incubated 1 h after heat shock without any treatment an those incubated with the combination ASA+H2O2 (n = 3) (p<0.001). C) RT-PCR of HSP-72 mRNA from PBMCs treated in vitro with the combination ASA+H2O2 following heat shock. Total RNA was obtained immediately after heat shock. Products of RT-PCR were resolve in agarose gels.
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pone-0065449-g002: Time course of changes in HSP-72 expression before (T0), immediately after heat shock (HS) and 1 (1 h) and 2 (2 h) hours after recovery at 37°C in peripheral blood mononuclear cells (PBMCs) from untreated rats incubated with or without the combination ASA (10 µM)+H2O2 (5 µM).A) Western blots of HSP-72 from rat PBMCs (100 µg of total protein) incubated with (+) or without (-) the indicated treatments for 1 h before heat shock, β-actin was used as loading control. Protein extracts were obtained before in vitro incubation (T0), or immediately after Heat shock (HS) or (1h) and 2 (2h) hours after recovery at 37°C. B) Histogram of the average signals expressed as Arbitrary units (AU) = OD for HSP-72/OD for β-actin from three independent experiments. *Comparison of HSP-72 content immediately after heat shock between cells without any treatment an those incubated with the combination ASA+H2O2 (n = 3) (p<0.001). **Comparison of cells incubated 1 h after heat shock without any treatment an those incubated with the combination ASA+H2O2 (n = 3) (p<0.001). C) RT-PCR of HSP-72 mRNA from PBMCs treated in vitro with the combination ASA+H2O2 following heat shock. Total RNA was obtained immediately after heat shock. Products of RT-PCR were resolve in agarose gels.

Mentions: Next we compared the priming effect of the ASA+H2O2 combination applied in vitro to that observed with the priming effect induced in vivo on PBMCs from ASA-challenged animals (45 mg/kg). The most significant effect was the accelerated expression of HSP-72 following in vitro heat shock when cells were pretreated with the ASA+H2O2 combination (lane 6 compared with lane 2 in Figure 2A). This accelerated expression of HSP-72 continued to be observed after 1 h following heat shock and disappearance after 2 h (Figure 2B).


HSP-72 accelerated expression in mononuclear cells induced in vivo by acetyl salicylic acid can be reproduced in vitro when combined with H2O2.

Sandoval-Montiel AA, Zentella-de-Piña M, Ventura-Gallegos JL, Frías-González S, López-Macay A, Zentella-Dehesa A - PLoS ONE (2013)

Time course of changes in HSP-72 expression before (T0), immediately after heat shock (HS) and 1 (1 h) and 2 (2 h) hours after recovery at 37°C in peripheral blood mononuclear cells (PBMCs) from untreated rats incubated with or without the combination ASA (10 µM)+H2O2 (5 µM).A) Western blots of HSP-72 from rat PBMCs (100 µg of total protein) incubated with (+) or without (-) the indicated treatments for 1 h before heat shock, β-actin was used as loading control. Protein extracts were obtained before in vitro incubation (T0), or immediately after Heat shock (HS) or (1h) and 2 (2h) hours after recovery at 37°C. B) Histogram of the average signals expressed as Arbitrary units (AU) = OD for HSP-72/OD for β-actin from three independent experiments. *Comparison of HSP-72 content immediately after heat shock between cells without any treatment an those incubated with the combination ASA+H2O2 (n = 3) (p<0.001). **Comparison of cells incubated 1 h after heat shock without any treatment an those incubated with the combination ASA+H2O2 (n = 3) (p<0.001). C) RT-PCR of HSP-72 mRNA from PBMCs treated in vitro with the combination ASA+H2O2 following heat shock. Total RNA was obtained immediately after heat shock. Products of RT-PCR were resolve in agarose gels.
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Related In: Results  -  Collection

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pone-0065449-g002: Time course of changes in HSP-72 expression before (T0), immediately after heat shock (HS) and 1 (1 h) and 2 (2 h) hours after recovery at 37°C in peripheral blood mononuclear cells (PBMCs) from untreated rats incubated with or without the combination ASA (10 µM)+H2O2 (5 µM).A) Western blots of HSP-72 from rat PBMCs (100 µg of total protein) incubated with (+) or without (-) the indicated treatments for 1 h before heat shock, β-actin was used as loading control. Protein extracts were obtained before in vitro incubation (T0), or immediately after Heat shock (HS) or (1h) and 2 (2h) hours after recovery at 37°C. B) Histogram of the average signals expressed as Arbitrary units (AU) = OD for HSP-72/OD for β-actin from three independent experiments. *Comparison of HSP-72 content immediately after heat shock between cells without any treatment an those incubated with the combination ASA+H2O2 (n = 3) (p<0.001). **Comparison of cells incubated 1 h after heat shock without any treatment an those incubated with the combination ASA+H2O2 (n = 3) (p<0.001). C) RT-PCR of HSP-72 mRNA from PBMCs treated in vitro with the combination ASA+H2O2 following heat shock. Total RNA was obtained immediately after heat shock. Products of RT-PCR were resolve in agarose gels.
Mentions: Next we compared the priming effect of the ASA+H2O2 combination applied in vitro to that observed with the priming effect induced in vivo on PBMCs from ASA-challenged animals (45 mg/kg). The most significant effect was the accelerated expression of HSP-72 following in vitro heat shock when cells were pretreated with the ASA+H2O2 combination (lane 6 compared with lane 2 in Figure 2A). This accelerated expression of HSP-72 continued to be observed after 1 h following heat shock and disappearance after 2 h (Figure 2B).

Bottom Line: Among NSAIDs acetyl salicylic acid remains as a valuable tool because of the variety of benefic prophylactic and therapeutic effects.This effect reaches its maximum 2 h after treatment and disappeared after 5 h.The fact that in vitro acetyl salicylic acid alone did not induce this priming effect implies that in vivo other signals are required.

View Article: PubMed Central - PubMed

Affiliation: Departmento de Medicina Genómica y Toxicología Ambiental, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, México, D.F., México.

ABSTRACT

Background: Among NSAIDs acetyl salicylic acid remains as a valuable tool because of the variety of benefic prophylactic and therapeutic effects. Nevertheless, the molecular bases for these responses have not been complete understood. We explored the effect of acetyl salicylic acid on the heat shock response.

Results: Peripheral blood mononuclear cells from rats challenged with acetyl salicylic acid presented a faster kinetics of expression of HSP-72 messenger RNA and protein in response to in vitro heat shock. This effect reaches its maximum 2 h after treatment and disappeared after 5 h. On isolated peripheral blood mononuclear cells from untreated rats, incubation with acetyl salicylic acid was ineffective to produce priming, but this effect was mimicked when the cells were incubated with the combination of H2O2+ ASA.

Conclusions: Administration of acetyl salicylic acid to rats alters HSP-72 expression mechanism in a way that it becomes more efficient in response to in vitro heat shock. The fact that in vitro acetyl salicylic acid alone did not induce this priming effect implies that in vivo other signals are required. Priming could be reproduces in vitro with the combination of acetyl salicylic acid+H2O2.

Show MeSH
Related in: MedlinePlus