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Aspirin influences megakaryocytic gene expression leading to up-regulation of multidrug resistance protein-4 in human platelets.

Massimi I, Guerriero R, Lotti LV, Lulli V, Borgognone A, Romani F, Barillà F, Gaudio C, Gabbianelli M, Frati L, Pulcinelli FM - Br J Clin Pharmacol (2014)

Bottom Line: We recently found that platelet MRP4 overexpression has a role in reducing aspirin action in patients after by-pass surgery.In DAMI cells, aspirin and WY14643 treatment induced a significant increase in MRP4 and PPARα expression.This work represents an innovative and attractive approach, useful both to identify patients less sensitive to aspirin and to improve pharmacological treatment in cardiovascular high-risk patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Experimental Medicine, Sapienza University of Rome, Rome.

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Related in: MedlinePlus

Aspirin stimulates endogenous MRP4 via a PPARα dependent mechanism in DAMI cells. Control non- specific siRNA (CTRsi) and PPARα specific siRNA (PPARsi) transfected DAMI cells were either untreated (Ctr) or treated, for 48 h, with either 50 μm aspirin (ASA) or 1 μm PPARα agonist, WY14643 (WY). (A) Q-RT-PCR analysis of endogenous PPARα expression. Data were normalized with the mean of the fold induction of GAPDH, ACTB and CD42B expression and reported as mean ± SD of three experiments (NS = not significant, **P < 0.01, ***P < 0.001; t-test). (B) Endogenous PPARα protein expression. Representative Western blot, out of three performed. (C) Q-RT-PCR analysis of endogenous MRP4 expression; Data were normalized with ACTB expression and reported as mean ± SD of three experiments (NS = not significant, **P < 0.01, ***P < 0.001; t-test)
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fig04: Aspirin stimulates endogenous MRP4 via a PPARα dependent mechanism in DAMI cells. Control non- specific siRNA (CTRsi) and PPARα specific siRNA (PPARsi) transfected DAMI cells were either untreated (Ctr) or treated, for 48 h, with either 50 μm aspirin (ASA) or 1 μm PPARα agonist, WY14643 (WY). (A) Q-RT-PCR analysis of endogenous PPARα expression. Data were normalized with the mean of the fold induction of GAPDH, ACTB and CD42B expression and reported as mean ± SD of three experiments (NS = not significant, **P < 0.01, ***P < 0.001; t-test). (B) Endogenous PPARα protein expression. Representative Western blot, out of three performed. (C) Q-RT-PCR analysis of endogenous MRP4 expression; Data were normalized with ACTB expression and reported as mean ± SD of three experiments (NS = not significant, **P < 0.01, ***P < 0.001; t-test)

Mentions: To confirm further the involvement of PPARα in MRP4 up-regulation, DAMI cells were transfected with PPARα specific siRNA (PPAR-si). Cells transfected with PPAR-si did not show any significant increase of PPARα mRNA and protein expression after aspirin and WY14643 treatment while cells transfected with control non-specific siRNA (CTR-si) showed the same aspirin and WY14643 induced expression changes observed in untrasfected cells (Figure 4A and B). Similarly, no increase of aspirin and WY14643 dependent MRP4 expression was detected in cells treated with PPAR-si, unlike those found in CTR-si transfected cells (Figure 4C).


Aspirin influences megakaryocytic gene expression leading to up-regulation of multidrug resistance protein-4 in human platelets.

Massimi I, Guerriero R, Lotti LV, Lulli V, Borgognone A, Romani F, Barillà F, Gaudio C, Gabbianelli M, Frati L, Pulcinelli FM - Br J Clin Pharmacol (2014)

Aspirin stimulates endogenous MRP4 via a PPARα dependent mechanism in DAMI cells. Control non- specific siRNA (CTRsi) and PPARα specific siRNA (PPARsi) transfected DAMI cells were either untreated (Ctr) or treated, for 48 h, with either 50 μm aspirin (ASA) or 1 μm PPARα agonist, WY14643 (WY). (A) Q-RT-PCR analysis of endogenous PPARα expression. Data were normalized with the mean of the fold induction of GAPDH, ACTB and CD42B expression and reported as mean ± SD of three experiments (NS = not significant, **P < 0.01, ***P < 0.001; t-test). (B) Endogenous PPARα protein expression. Representative Western blot, out of three performed. (C) Q-RT-PCR analysis of endogenous MRP4 expression; Data were normalized with ACTB expression and reported as mean ± SD of three experiments (NS = not significant, **P < 0.01, ***P < 0.001; t-test)
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig04: Aspirin stimulates endogenous MRP4 via a PPARα dependent mechanism in DAMI cells. Control non- specific siRNA (CTRsi) and PPARα specific siRNA (PPARsi) transfected DAMI cells were either untreated (Ctr) or treated, for 48 h, with either 50 μm aspirin (ASA) or 1 μm PPARα agonist, WY14643 (WY). (A) Q-RT-PCR analysis of endogenous PPARα expression. Data were normalized with the mean of the fold induction of GAPDH, ACTB and CD42B expression and reported as mean ± SD of three experiments (NS = not significant, **P < 0.01, ***P < 0.001; t-test). (B) Endogenous PPARα protein expression. Representative Western blot, out of three performed. (C) Q-RT-PCR analysis of endogenous MRP4 expression; Data were normalized with ACTB expression and reported as mean ± SD of three experiments (NS = not significant, **P < 0.01, ***P < 0.001; t-test)
Mentions: To confirm further the involvement of PPARα in MRP4 up-regulation, DAMI cells were transfected with PPARα specific siRNA (PPAR-si). Cells transfected with PPAR-si did not show any significant increase of PPARα mRNA and protein expression after aspirin and WY14643 treatment while cells transfected with control non-specific siRNA (CTR-si) showed the same aspirin and WY14643 induced expression changes observed in untrasfected cells (Figure 4A and B). Similarly, no increase of aspirin and WY14643 dependent MRP4 expression was detected in cells treated with PPAR-si, unlike those found in CTR-si transfected cells (Figure 4C).

Bottom Line: We recently found that platelet MRP4 overexpression has a role in reducing aspirin action in patients after by-pass surgery.In DAMI cells, aspirin and WY14643 treatment induced a significant increase in MRP4 and PPARα expression.This work represents an innovative and attractive approach, useful both to identify patients less sensitive to aspirin and to improve pharmacological treatment in cardiovascular high-risk patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Experimental Medicine, Sapienza University of Rome, Rome.

Show MeSH
Related in: MedlinePlus