Inhibitory Potencies of Several Iridoids on Cyclooxygenase-1, Cyclooxygnase-2 Enzymes Activities, Tumor Necrosis Factor-α and Nitric Oxide Production In Vitro

Abstract

To verify the anti-inflammatory potency of iridoids, seven iridoid glucosides (aucubin, catalpol, gentiopicroside, swertiamarin, geniposide, geniposidic acid and loganin) and an iridoid aglycone (genipin) were investigated with in vitro testing model systems based on inhibition of cyclooxygenase (COX)-1/-2 enzymes, the tumor necrosis factor-α (TNF-α) formation and nitric oxide (NO) production. The hydrolyzed-iridoid products (H-iridoid) with β-gludosidase treatment only showed inhibitory activities, and revealed different potencies, depending on their chemical structures. Without the β-gludosidase treatment, no single iridoid glycoside exhibited any activities. The aglycone form (genipin) also did not show inhibitory activities. To compare anti-inflammatory potency, the inhibitory concentrations (IC₅₀) in each testing system were measured. The hydrolyzed-aucubin product (H-aucubin) with β-gludosidase treatment showed a moderate inhibition on COX-2 with IC₅₀ of 8.83 μM, but much less inhibition (IC₅₀, 68.9 μM) on COX-1 was noted. Of the other H-iridoid products, the H-loganin and the H-geniposide exhibited higher inhibitory effects on COX-1, revealing IC₅₀ values of 3.55 and 5.37 μM, respectively. In the case of TNF-α assay, four H-iridoid products: H-aucubin, H-catalpol, H-geniposide and H-loganin suppressed the TNF-α formation with IC₅₀ values of 11.2, 33.3, 58.2 and 154.6 μM, respectively. But other H-iridoid products manifested no significant activity. Additional experiments on NO production were conducted. We observed that only the H-aucubin exhibited a significant suppression with IC₅₀ value of 14.1 μM. Genipin, an agycone form, showed no inhibitory effects on all testing models, implying the hydrolysis of the glycosidic bond of iridoid glycoside is a pre-requisite step to produce various biological activities.