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Aspirin Action in Endothelial Cells: Different Patterns of Response Between Chemokine CX3CL1/CX3CR1 and TNF-α/TNFR1 Signaling Pathways.

Szukiewicz D, Wojciechowska M, Bilska A, Stangret A, Szewczyk G, Mittal TK, Watroba M, Kochanowski J - Cardiovasc Drugs Ther (2015)

Bottom Line: We examined the effects of aspirin on CX3CL1 and TNF-α production, as well as CX3CR1 and TNFR1 expression.Aspirin significantly (p <  .05) decreased CX3CL1 production, and the mean decrease in CX3CL1 production was inversely proportional to increased (p < 0.05) expression of CX3CR1.Autoregulation between CX3CL1 and CX3CR1 may explain overexpression of CX3CR1 as the compensatory effect in aspirin-treated HUVECs.

View Article: PubMed Central - PubMed

Affiliation: Department of General & Experimental Pathology with Centre for Preclinical Research and Technology (CEPT), Medical University of Warsaw, ul. Pawinskiego 3C, 02-106, Warsaw, Poland, Dszukiewicz@hotmail.com.

ABSTRACT

Purpose: TNF-α induces fractalkine (CX3CL1) and its receptor CX3CR1 in endothelial cells through NF-қB activation. NF-қB inhibitors may reduce the expression of CX3CL1, and modulation of the CX3CL1/CX3CR1 signaling was proposed as a new target for aspirin. We examined the effects of aspirin on CX3CL1 and TNF-α production, as well as CX3CR1 and TNFR1 expression.

Methods: HUVECs isolated after term pregnancies (N = 28) were cultured in vitro. Lipopolysaccharide (1 μg/ml) was used as CX3CL1 inducer. HUVECs were exposed to six different concentrations of aspirin (between 1.0 and 6.0 mM) during 7 days. The levels of CX3CL1 and TNF-α in the culture media were measured using ELISA. After termination of the cultures, mean expressions of CX3CR1 and TNFR1 were examined in the immunostained paraffin sections using quantitative immunohistochemistry.

Results: Aspirin significantly (p <  .05) decreased CX3CL1 production, and the mean decrease in CX3CL1 production was inversely proportional to increased (p < 0.05) expression of CX3CR1. The combined mean CX3CL1 concentrations, including all time points, equaled 782.18 ± 74.4 pg/ml in aspirin treated HUVECs compared to a total concentration of 2467.53 ± 127.5 pg/ml combined from the respective time points in the controls. An inhibition of TNF-α production in HUVECs after pretreatment with aspirin was observed. Unlike in the case of CX3CR1 expression, there were no signs of TNFR1 upregulation.

Conclusions: Autoregulation between CX3CL1 and CX3CR1 may explain overexpression of CX3CR1 as the compensatory effect in aspirin-treated HUVECs. Inhibition of CX3CR1 could prevent thrombotic complications in the early period after discontinuation of aspirin.

No MeSH data available.


Related in: MedlinePlus

Cell viability assay of HUVECs exposured to different doses of aspirin over time (at a given time points)
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Fig2: Cell viability assay of HUVECs exposured to different doses of aspirin over time (at a given time points)

Mentions: Aspirin treatment produced a significant (p < 0.05) decrease in the mean concentration of CX3CL1 in the HUVECs culture supernatant, especially when administered at doses of 2.0 and 3.0 mM (Fig. 1, Tables 2 and 3). This curtailment of CX3CL1 production persisted throughout the whole period of observation (time points 24, 48, 72, and 144 h). The combined mean CX3CL1 concentration for group II and group III, including all time points, equaled 782.18 ± 74.4 pg/ml compared to a total concentration of 2467.53 ± 127.5 pg/ml combined from the respective time points in group VII (control). Analysis of the cell viability in the HUVEC cultures revealed that the highest tested dose of aspirin (6.0 mM) resulted in a significant increase in cytotoxicity (Fig. 2). This effect was observed at the beginning (24 h after aspirin administration), and the mean cell viability still lingered below the mean values obtained in the controls (p < 0.05) at the respective time points.Fig. 1


Aspirin Action in Endothelial Cells: Different Patterns of Response Between Chemokine CX3CL1/CX3CR1 and TNF-α/TNFR1 Signaling Pathways.

Szukiewicz D, Wojciechowska M, Bilska A, Stangret A, Szewczyk G, Mittal TK, Watroba M, Kochanowski J - Cardiovasc Drugs Ther (2015)

Cell viability assay of HUVECs exposured to different doses of aspirin over time (at a given time points)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4522030&req=5

Fig2: Cell viability assay of HUVECs exposured to different doses of aspirin over time (at a given time points)
Mentions: Aspirin treatment produced a significant (p < 0.05) decrease in the mean concentration of CX3CL1 in the HUVECs culture supernatant, especially when administered at doses of 2.0 and 3.0 mM (Fig. 1, Tables 2 and 3). This curtailment of CX3CL1 production persisted throughout the whole period of observation (time points 24, 48, 72, and 144 h). The combined mean CX3CL1 concentration for group II and group III, including all time points, equaled 782.18 ± 74.4 pg/ml compared to a total concentration of 2467.53 ± 127.5 pg/ml combined from the respective time points in group VII (control). Analysis of the cell viability in the HUVEC cultures revealed that the highest tested dose of aspirin (6.0 mM) resulted in a significant increase in cytotoxicity (Fig. 2). This effect was observed at the beginning (24 h after aspirin administration), and the mean cell viability still lingered below the mean values obtained in the controls (p < 0.05) at the respective time points.Fig. 1

Bottom Line: We examined the effects of aspirin on CX3CL1 and TNF-α production, as well as CX3CR1 and TNFR1 expression.Aspirin significantly (p <  .05) decreased CX3CL1 production, and the mean decrease in CX3CL1 production was inversely proportional to increased (p < 0.05) expression of CX3CR1.Autoregulation between CX3CL1 and CX3CR1 may explain overexpression of CX3CR1 as the compensatory effect in aspirin-treated HUVECs.

View Article: PubMed Central - PubMed

Affiliation: Department of General & Experimental Pathology with Centre for Preclinical Research and Technology (CEPT), Medical University of Warsaw, ul. Pawinskiego 3C, 02-106, Warsaw, Poland, Dszukiewicz@hotmail.com.

ABSTRACT

Purpose: TNF-α induces fractalkine (CX3CL1) and its receptor CX3CR1 in endothelial cells through NF-қB activation. NF-қB inhibitors may reduce the expression of CX3CL1, and modulation of the CX3CL1/CX3CR1 signaling was proposed as a new target for aspirin. We examined the effects of aspirin on CX3CL1 and TNF-α production, as well as CX3CR1 and TNFR1 expression.

Methods: HUVECs isolated after term pregnancies (N = 28) were cultured in vitro. Lipopolysaccharide (1 μg/ml) was used as CX3CL1 inducer. HUVECs were exposed to six different concentrations of aspirin (between 1.0 and 6.0 mM) during 7 days. The levels of CX3CL1 and TNF-α in the culture media were measured using ELISA. After termination of the cultures, mean expressions of CX3CR1 and TNFR1 were examined in the immunostained paraffin sections using quantitative immunohistochemistry.

Results: Aspirin significantly (p <  .05) decreased CX3CL1 production, and the mean decrease in CX3CL1 production was inversely proportional to increased (p < 0.05) expression of CX3CR1. The combined mean CX3CL1 concentrations, including all time points, equaled 782.18 ± 74.4 pg/ml in aspirin treated HUVECs compared to a total concentration of 2467.53 ± 127.5 pg/ml combined from the respective time points in the controls. An inhibition of TNF-α production in HUVECs after pretreatment with aspirin was observed. Unlike in the case of CX3CR1 expression, there were no signs of TNFR1 upregulation.

Conclusions: Autoregulation between CX3CL1 and CX3CR1 may explain overexpression of CX3CR1 as the compensatory effect in aspirin-treated HUVECs. Inhibition of CX3CR1 could prevent thrombotic complications in the early period after discontinuation of aspirin.

No MeSH data available.


Related in: MedlinePlus