The inhibition of inducible nitric oxide production in lipopolysaccharide-stimulated rat macrophages and in silico studies by flavonoids from Iris spuria L. rhizomes

Abstract

Ethnopharmacological relevanceIris is the largest genus in the family Iridaceae. Iris plants are distributed in tropical regions of the world. They are used as ornamentals and traditionally used to treat a variety of ailments. AimThis study aimed to evaluate the anti-inflammatory effect of flavonoids isolated from Iris spuria L. Materials and methodsThe isolated flavonoids (1-4) were identified on the basis of different spectroscopic methods (1D- and 2D-NMR) and co-TLC with authentic samples. The anti-inflammatory effect was tested on lipopolysaccharide (LPS)-induced nitric oxide (NO) production from rat-isolated peritoneal macrophages. Modeling and docking simulations of the compounds were performed using Molecular Operating Environment software and the crystal structure of the murine inducible nitric oxide synthase (iNOS). ResultsFour flavonoids (1-4) had been isolated from the rhizomes of Iris spuria L. (Hocka Hoona) for the first time. They were characterized as 5,7,2'-trihydroxy-6-methoxyflavanone (1), tectorigenin 7-O-β-D-glucopyranoside (2), tectorigenin 4'-O-β-D-glucopyranoside (3), and tectorigenin 4'-O-[β-D-glucopyranosyl(1 → 6)-β-D-glucopyranoside] (4). The selective inducible NO synthase inhibitor; aminoguanidine was used as a positive control. The production of nitric oxide (NO) was inhibited in a dose-dependent manner of the isolated compounds along with isoflavonoids (5-9) previously isolated from Iris spuria L. (Calizona). A concentration of 60 μg/ml of all tested compounds showed a significant inhibitory effect compared to media with LPS. Molecular modeling experiments supported the obtained biological data. ConclusionOur results reveal that flavonoids isolated from I. spuria L. (Hocka Hoona) and I. spuria L. (Calizona) appear to have a potential anti-inflammatory effect via inhibition of iNOS.