CULTURED HUMAN SYNOVIAL FIBROBLASTS RAPIDLY METABOLIZE KININS AND NEUROPEPTIDES

Kinins and substance P have been implicated in the pathogenesis sis of inflammatory arthritis by virtue of their abilities to induce vasodilation, edema, and pain. The relative biological potencies of these peptides in vivo would depend at least in part upon their rates of catabolism in the joint. We hypothesized that human synovial lining cells may regulate intraarticular levels of kinins and neuropeptides via degradation by cell surface-associated peptidases. We exposed intact human synovial fibroblasts to kinins and substance P, in the presence or absence of specific peptidase inhibitors, and measured the amount of intact substrate remaining and degradation product(s) generated over time. Aminopeptidase M (AmM; EC 3.4.11.2), neutral endopeptidase-24.11 (NEP-24.11; EC 3.4.24.11), and dipeptidyl(amino)peptidase IV (DAP IV; EC 3.4.14.5) were identified on the cell surface of synovial cells. Bradykinin degradation was due entirely to NEP-24.11 (1.39+/-0.29 nmol/min per well). Lysylbradykinin was also degraded by NEP-24.11 (0.80+/-0.19 nmol/min per well); however, in the presence of phosphoramidon, AmM-mediated conversion to bradykinin (3.74+/-0.46 nmol/min per well) could be demonstrated. The combined actions of NEP-24.11 (0.93+/-0.15 nmol/min per well) and DAP IV (0.84+/-0.18 nmol/min per well) were responsible for the degradation of substance P. AmM (2.44+/-0.33 nmol/min per well) and NEP-24.11 (1.30+/-0.45 nmol/min per well) were responsible for the degradation of the opioid peptide, [Leu5]enkephalin. The identity of each of the three peptidases was confirmed via synthetic substrate hydrolysis, inhibition profile, and immunological identification. The profiles of peptidase enzymes identified in cells derived from rheumatoid and osteoarthritic joints were identical. These data demonstrate the human synovial fibroblast to be a rich source of three specific peptidases and suggest that it may play a prominent role in regulating peptide levels in the joint.