title: alpha 1-Proteinase inhibitor in gingival crevicular fluid of humans with adult periodontitis: serpinolytic inhibition by doxycycline.
abstract: The serum protein, alpha 1-proteinase inhibitor (alpha 1-PI), defends the host against serine proteinases, e.g. PMN elastase. Using a rabbit anti-serum against human alpha 1-PI, this protein in GCF was quantified from a standard curve constructed from dot-blot analysis and characterized by Western blot. GCF was collected on filter paper strips from healthy (H), gingivitis (G) and adult periodontitis (AP) patients, then extracted with Tris/NaCl/CaCl2 buffer, pH 7.6. alpha 1-PI concentration increased with G and was highest in AP subjects. H sites only showed intact alpha 1-PI (52 kDa); no degradation fragments (48 kDa) were detected. In G and AP subjects, alpha 1-PI degradation fragments were seen in 17% and 71% of GCF samples, respectively. Both collagenase and alpha 1-PI-degrading activities in GCF increased with severity of inflammation (GCF flow). Moreover, the alpha 1-PI degrading (or serpinolytic) activity was characterized as a matrix metalloproteinase, probably collagenase, based on its in vitro response to a panel of different proteinase inhibitors including doxycycline. We propose: (1) that collagenase promotes periodontal breakdown not only by degrading collagen, but also by depleting alpha 1-PI regulation of elastase and other serine-proteinases, thereby favoring a broader attack on extracellular matrix (ECM) constituents, and (2) based on a recent longitudinal double-blind study using the techniques described above for alpha 1-PI analysis, that low-dose doxycycline administration to humans with adult periodontitis can inhibit this broad cascade of ECM degradation.
authors.affiliation: Department of Oral Biology and Pathology, School of Dental Medicine, SUNY at Stony Brook 11794, USA.
authors.forename: H M
authors.initials: HM
authors.lastname: Lee
authors.forename: L M
authors.initials: LM
authors.lastname: Golub
authors.forename: D
authors.initials: D
authors.lastname: Chan
authors.forename: M
authors.initials: M
authors.lastname: Leung
authors.forename: K
authors.initials: K
authors.lastname: Schroeder
authors.forename: M
authors.initials: M
authors.lastname: Wolff
authors.forename: S
authors.initials: S
authors.lastname: Simon
authors.forename: R
authors.initials: R
authors.lastname: Crout
mesh_terms: D000328:Adult; D000704:Analysis of Variance; D000900:Anti-Bacterial Agents / Q000378:metabolism*; D017364:Collagenases / Q000378:metabolism; D004318:Doxycycline / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005883:Gingival Crevicular Fluid / Q000201:enzymology*; D006801:Humans; D015151:Immunoblotting; D019272:Leukocyte Elastase / Q000378:metabolism; D008297:Male; D009504:Neutrophils / Q000201:enzymology; D010510:Periodontal Diseases / Q000201:enzymology*; D014361:Trypsin Inhibitors / Q000032:analysis / Q000378:metabolism*; D000515:alpha 1-Antitrypsin / Q000032:analysis / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D014361:Trypsin Inhibitors; D000515:alpha 1-Antitrypsin; D019272:Leukocyte Elastase; D017364:Collagenases; D004318:Doxycycline