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Involvement of p38 MAPK in the up-regulation of tissue factor on endothelial cells by antiphospholipid antibodies.
Arthritis Rheum. 2005 May; 52(5):1545-54.AR

Abstract

OBJECTIVE

To study the intracellular mechanism involved in the up-regulation of tissue factor (TF) on endothelial cells (ECs) by antiphospholipid antibodies (aPL), we examined the effects of aPL on the transcription, expression, and function of TF, the expression of interleukin-6 (IL-6) and IL-8, the induction of inducible nitric oxide synthase (iNOS), and the phosphorylation of p38 MAPK on human umbilical vein ECs (HUVECs).

METHODS

Cultured HUVECs were treated with IgG aPL (from patients with antiphospholipid syndrome [APS]) or with control IgG (from normal human serum). Phorbol myristate acetate (PMA) and bacterial lipopolysaccharide (LPS) were used as positive controls. TF expression was determined on the surface of HUVECs using an enzyme-linked immunosorbent assay (ELISA). TF activity was determined with the use of a chromogenic assay in cell lysates, and TF messenger RNA (mRNA) was determined by real-time quantitative polymerase chain reaction. Phosphorylation of p38 MAPK and induction of iNOS were determined by Western blotting, and levels of IL-6 and IL-8 were determined by ELISA.

RESULTS

PMA, LPS, and aPL significantly increased the expression of TF compared with controls. This up-regulation was significantly inhibited by SB203580 (a specific inhibitor of p38 MAPK) and by MG132 (a specific inhibitor of NF-kappaB). TF activity was significantly increased by treatment with IgG aPL and this effect was also inhibited by SB203580. Incubation of HUVECs with aPL increased TF mRNA 2-15-fold; these effects were abrogated by SB203580. IgG aPL induced significant phosphorylation of p38 MAPK and produced iNOS on HUVECs in a time-dependent manner. Treatment with IgG aPL also induced increased expression of IL-6 and IL-8 on HUVECs.

CONCLUSION

Our data show that aPL induces significant increases in TF transcription, function, and expression, in IL-6 and IL-8 up-regulation, and in iNOS expression on HUVECs and that these processes involve phosphorylation of p38 MAPK and activation of NF-kappaB. Understanding intracellular events in aPL-mediated EC activation may help in designing new targeted therapies for thrombosis in APS.

Authors+Show Affiliations

Morehouse School of Medicine, Atlanta, Georgia, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

15880836

Citation

Vega-Ostertag, Mariano, et al. "Involvement of P38 MAPK in the Up-regulation of Tissue Factor On Endothelial Cells By Antiphospholipid Antibodies." Arthritis and Rheumatism, vol. 52, no. 5, 2005, pp. 1545-54.
Vega-Ostertag M, Casper K, Swerlick R, et al. Involvement of p38 MAPK in the up-regulation of tissue factor on endothelial cells by antiphospholipid antibodies. Arthritis Rheum. 2005;52(5):1545-54.
Vega-Ostertag, M., Casper, K., Swerlick, R., Ferrara, D., Harris, E. N., & Pierangeli, S. S. (2005). Involvement of p38 MAPK in the up-regulation of tissue factor on endothelial cells by antiphospholipid antibodies. Arthritis and Rheumatism, 52(5), 1545-54.
Vega-Ostertag M, et al. Involvement of P38 MAPK in the Up-regulation of Tissue Factor On Endothelial Cells By Antiphospholipid Antibodies. Arthritis Rheum. 2005;52(5):1545-54. PubMed PMID: 15880836.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Involvement of p38 MAPK in the up-regulation of tissue factor on endothelial cells by antiphospholipid antibodies. AU - Vega-Ostertag,Mariano, AU - Casper,Katherine, AU - Swerlick,Robert, AU - Ferrara,Dardo, AU - Harris,E Nigel, AU - Pierangeli,Silvia S, PY - 2005/5/10/pubmed PY - 2005/6/30/medline PY - 2005/5/10/entrez SP - 1545 EP - 54 JF - Arthritis and rheumatism JO - Arthritis Rheum VL - 52 IS - 5 N2 - OBJECTIVE: To study the intracellular mechanism involved in the up-regulation of tissue factor (TF) on endothelial cells (ECs) by antiphospholipid antibodies (aPL), we examined the effects of aPL on the transcription, expression, and function of TF, the expression of interleukin-6 (IL-6) and IL-8, the induction of inducible nitric oxide synthase (iNOS), and the phosphorylation of p38 MAPK on human umbilical vein ECs (HUVECs). METHODS: Cultured HUVECs were treated with IgG aPL (from patients with antiphospholipid syndrome [APS]) or with control IgG (from normal human serum). Phorbol myristate acetate (PMA) and bacterial lipopolysaccharide (LPS) were used as positive controls. TF expression was determined on the surface of HUVECs using an enzyme-linked immunosorbent assay (ELISA). TF activity was determined with the use of a chromogenic assay in cell lysates, and TF messenger RNA (mRNA) was determined by real-time quantitative polymerase chain reaction. Phosphorylation of p38 MAPK and induction of iNOS were determined by Western blotting, and levels of IL-6 and IL-8 were determined by ELISA. RESULTS: PMA, LPS, and aPL significantly increased the expression of TF compared with controls. This up-regulation was significantly inhibited by SB203580 (a specific inhibitor of p38 MAPK) and by MG132 (a specific inhibitor of NF-kappaB). TF activity was significantly increased by treatment with IgG aPL and this effect was also inhibited by SB203580. Incubation of HUVECs with aPL increased TF mRNA 2-15-fold; these effects were abrogated by SB203580. IgG aPL induced significant phosphorylation of p38 MAPK and produced iNOS on HUVECs in a time-dependent manner. Treatment with IgG aPL also induced increased expression of IL-6 and IL-8 on HUVECs. CONCLUSION: Our data show that aPL induces significant increases in TF transcription, function, and expression, in IL-6 and IL-8 up-regulation, and in iNOS expression on HUVECs and that these processes involve phosphorylation of p38 MAPK and activation of NF-kappaB. Understanding intracellular events in aPL-mediated EC activation may help in designing new targeted therapies for thrombosis in APS. SN - 0004-3591 UR - https://www.unboundmedicine.com/medline/citation/15880836/Involvement_of_p38_MAPK_in_the_up_regulation_of_tissue_factor_on_endothelial_cells_by_antiphospholipid_antibodies_ L2 - https://doi.org/10.1002/art.21009 DB - PRIME DP - Unbound Medicine ER -