Studies in anti-inflammatory effect of spinorphin, an endogenous factor

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It has been known that spinorphin has the inhibitory activity toward various enkephalin-degrading enzymes, such as neutral endopeptidase, aminopeptidase and dipeptidyl peptidase III (DPPIII) in central nerve system. In this study, the inhibitory activity toward DPPIII on human PMNs was found using a specific substrate (Arg-Arg-β-naphtylamide) and inhibitors.To characterize anti-inflammatory effect of spinorphin, an endogenous peptide purified from bovine spinal cord, priming of spinorphin on human neutrophils (PMNs) and quantitative method of spinorphin in human serum were investigates.(1) Priming effect of superoxide generation by stimulant-induced PMNsSpinorphin (1μM) alone did not induce superoxide generation. However, by secondary stimulation with FMLP, an inflammatory stimulant, the activity was markedly augmented compared to that of nontreated control. From FACS analysis, expression of CD11b and CD18, adhesion molecules was significantly increased compared with that of control, suggesting that the priming effect of spinorphin is in part due to increase of expression of adhesion molecules.(2) Effect of apinorphin on dipeptidyl peptidase III enzymatic activity(3) Quantitative method of spinorphin by ELISAAnti-spinorphin antibody was produced in rabbits immunized with keyhole limpet hemocyanin (KLH)-conjugated spinorphin. Quantitative method of spinorphin by ELISA was established. Anti-spinorphin antibody did not complete with its analogues in this method. Base levels of spinorphin in human serum were 0.23±0.25 pmol/ml.From these results, spinorphin may be a peptide regulator in inflammation.