title: Effect of Low Molecular Weight Heparins (LMWHs) on antiphospholipid Antibodies (aPL)-mediated inhibition of endometrial angiogenesis.
abstract: Antiphospholipid syndrome (APS) is an autoimmune disorder characterized by vascular thrombosis and/or pregnancy morbidity in the presence of circulating antiphospholipid antibodies (aPL). Different pathogenic mechanisms for aPL-mediated pregnancy failure have been proposed. In particular a direct effect of aPL on both maternal and fetal side of the placental tissue has been reported, since their reactivity with β2-glycoprotein I (β2GPI) makes them adhere to trophoblast and human endometrial endothelial cell (HEEC) membranes. β2GPI can be recognized by aPL that, once bound, interfere with both trophoblast functions and with the HEEC differentiation.APS patients can be successfully treated with Low Molecular Weight Heparin (LMWH). Recent reports suggest that LMWH acts through mechanisms alternative to its well known anticoagulant effect, because of its ability to bind β2GPI. In our previous studies, we showed that LMWH is able to reduce the aPL binding to trophoblasts and restore cell invasiveness and differentiation. So far, however, no study has described its effects on endometrial angiogenesis.The aim of our research was to evaluate whether two LMWHs, tinzaparin and enoxaparin, have an effect on the aPL-inhibited endometrial angiogenesis. This prompted us to investigate: (i) in vitro HEEC angiogenesis through a Matrigel assay; (ii) VEGF secretion by ELISA; (iii) matrix metalloproteinase-2 (MMP-2) activity by gelatin zymography; (iv) Nuclear Factor-κB (NF-κB) DNA binding activity by colorimetric assay; (v) STAT-3 activation by a sandwich-ELISA kit. Furthermore, using an in vivo murine model we investigated the LMWHs effects on angiogenesis.We demonstrated that the addition of LMWHs prevents aPL-inhibited HEEC angiogenesis, both in vitro and in vivo, and is able to restore the aPL inhibited NF-κB and/or STAT-3 activity, the VEGF secretion and the MMPs activity.The demonstration of a beneficial role for LMWHs on the aPL-inhibited HEEC angiogenesis might provide additional mechanisms whereby this treatment protects early pregnancy in APS.
authors.affiliation: Department of Obstetrics and Gynecology, Università Cattolica del Sacro Cuore, Rome, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: D'Ippolito
authors.forename: Riccardo
authors.initials: R
authors.lastname: Marana
authors.forename: Fiorella
authors.initials: F
authors.lastname: Di Nicuolo
authors.forename: Roberta
authors.initials: R
authors.lastname: Castellani
authors.forename: Manuela
authors.initials: M
authors.lastname: Veglia
authors.forename: John
authors.initials: J
authors.lastname: Stinson
authors.forename: Giovanni
authors.initials: G
authors.lastname: Scambia
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Di Simone
mesh_terms: D000328:Adult; D000818:Animals; D017152:Antibodies, Antiphospholipid / Q000494:pharmacology*; D016736:Antiphospholipid Syndrome / Q000517:prevention & control; D004305:Dose-Response Relationship, Drug; D004717:Endometrium / Q000098:blood supply* / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D017984:Enoxaparin / Q000494:pharmacology*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006495:Heparin, Low-Molecular-Weight / Q000494:pharmacology*; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D050796:STAT3 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D000078222:Tinzaparin; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D017152:Antibodies, Antiphospholipid; D017984:Enoxaparin; D006495:Heparin, Low-Molecular-Weight; D016328:NF-kappa B; D050796:STAT3 Transcription Factor; D042461:Vascular Endothelial Growth Factor A; D000078222:Tinzaparin; D020778:Matrix Metalloproteinase 2