title: Recombinant TFPI-2 enhances macrophage apoptosis through upregulation of Fas/FasL.
abstract: Tissue factor pathway inhibitor-2 (TFPI-2) is a Kunitz-type serine proteinase inhibitor with inhibitory activity toward activated factor XI, plasma kallikrein, plasmin, certain matrix metalloproteinases, and the tissue factor-activated factor VII complex. In addition, TFPI-2 has other functions such as promoting cell migration and inducing apoptosis. In the present study, we investigated if TFPI-2 induced apoptosis in cultured U937-derived macrophages and the possible signal pathways that involved in the apoptotic process. Apoptotic DNA fragment detection and caspase-3,9 activity measurements indicated that rTFPI-2 promoted U937-derived macrophage apoptosis. Hoechst 33342 assay and flow cytometry further showed that rTFPI-2 induced apoptosis in cultured macrophages in a dose-dependent manner. Because death receptors of the TNF family such as Fas are the best-understood death pathways that recruit Fas-associated death domain (FADD) and procaspase-8 to the receptor in macrophages, we investigated the expression of Fas and its ligand (FasL) and downstream signal caspase-8 by Western blot analysis. The results indicated that the process of apoptosis triggered by rTFPI-2 was, at least in part, actively conducted by U937-derived macrophages possibly through Fas/FasL signal pathway. In brief, rTFPI-2 may have the potential usefulness in inducing macrophages apoptosis, which suggest TFPI-2 might have antiatherogenic effects.
authors.affiliation: Department of Cardiovascular Medicine, Huashan Hospital Affiliated to Fudan University, Shanghai, 200040, China.
authors.forename: Jun-Jie
authors.initials: JJ
authors.lastname: Pan
authors.forename: Hai-Ming
authors.initials: HM
authors.lastname: Shi
authors.forename: Xin-Ping
authors.initials: XP
authors.lastname: Luo
authors.forename: Duan
authors.initials: D
authors.lastname: Ma
authors.forename: Yong
authors.initials: Y
authors.lastname: Li
authors.forename: Jun
authors.initials: J
authors.lastname: Zhu
authors.forename: Wang
authors.initials: W
authors.lastname: Liang
authors.forename: Jin-Gui
authors.initials: JG
authors.lastname: Mu
authors.forename: Jian
authors.initials: J
authors.lastname: Li
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D001562:Benzimidazoles; D020169:Caspases / Q000378:metabolism; D053938:DNA Fragmentation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D053222:Fas Ligand Protein / Q000378:metabolism*; D053200:Fas-Associated Death Domain Protein / Q000378:metabolism; D005456:Fluorescent Dyes; D006023:Glycoproteins / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D008264:Macrophages / Q000187:drug effects* / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D015842:Serine Proteinase Inhibitors / Q000378:metabolism / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D014407:Tumor Cells, Cultured; D020298:U937 Cells; D015854:Up-Regulation; D019014:fas Receptor / Q000378:metabolism*
chemical_list: D001562:Benzimidazoles; D053222:Fas Ligand Protein; D053200:Fas-Associated Death Domain Protein; D005456:Fluorescent Dyes; D006023:Glycoproteins; D011994:Recombinant Proteins; D015842:Serine Proteinase Inhibitors; D019014:fas Receptor; C021240:tissue-factor-pathway inhibitor 2; D020169:Caspases; C017807:bisbenzimide ethoxide trihydrochloride