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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

Relationship between aspirin dose and the mean NF-қB (NF-қB/p65 protein heterodimer) concentrations in HUVECs cell lysates. The cultures were subjected to LPS pretreatment (1 μg/ml)
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Fig6: Relationship between aspirin dose and the mean NF-қB (NF-қB/p65 protein heterodimer) concentrations in HUVECs cell lysates. The cultures were subjected to LPS pretreatment (1 μg/ml)

Mentions: The levels of NF-қB/p65 reflected augmented production of these proteins in HUVECs under the influence of LPS, a known NF-қB inducer, administered initially (24 h after the cells were plated). Examination of the HUVEC cell lysates revealed a negative correlation between the administered dose of aspirin and the NF-қB/p65 protein level (Fig. 6). In groups III, IV, V, and VI (aspirin doses 3.0, 4.0, 5.0, and 6.0 mM, respectively), NF-қB (NF-қB/p65 protein heterodimer) concentrations were significantly (p < 0.05) lower (approximately 2-fold to 3.4-fold) compared to the control group.Fig. 6


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)

Relationship between aspirin dose and the mean NF-қB (NF-қB/p65 protein heterodimer) concentrations in HUVECs cell lysates. The cultures were subjected to LPS pretreatment (1 μg/ml)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig6: Relationship between aspirin dose and the mean NF-қB (NF-қB/p65 protein heterodimer) concentrations in HUVECs cell lysates. The cultures were subjected to LPS pretreatment (1 μg/ml)
Mentions: The levels of NF-қB/p65 reflected augmented production of these proteins in HUVECs under the influence of LPS, a known NF-қB inducer, administered initially (24 h after the cells were plated). Examination of the HUVEC cell lysates revealed a negative correlation between the administered dose of aspirin and the NF-қB/p65 protein level (Fig. 6). In groups III, IV, V, and VI (aspirin doses 3.0, 4.0, 5.0, and 6.0 mM, respectively), NF-қB (NF-қB/p65 protein heterodimer) concentrations were significantly (p < 0.05) lower (approximately 2-fold to 3.4-fold) compared to the control group.Fig. 6

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