Inhibition of Nitric Oxide Production by Ethyl Digallates Isolated from Galla Rhois in RAW 264.7 Macrophages

Abstract

Galla Rhois and its components are known to possess anti-infl ammatory properties. In the present study, we prepared equilibrium mixture of ethyl m-digallate and ethyl p-digallate isomers (EDG) from Galla Rhois and examined its effect on nitric oxide (NO) production in murine macrophage cell line. Treatment of RAW264.7 macrophages with EDG signifi cantly inhibited NO production and inducible nitric oxide synthase (iNOS) expression stimulated by LPS, as assessed by Western blot and quantitative RT-PCR analyses. We also demonstrated that EDG treatment led to an increase in heme oxygenase-1 (HO-1) mRNA and protein expression. EDG treatment also enhanced expression level of nuclear factor-erythroid 2-related factor 2 (Nrf2) in nucleus, which is critical for transcriptional induction of HO-1. Treatment with SnPP (tin protoporphyrin IX), a selective HO-1 inhibitor, reversed EDG-mediated inhibition of nitrite production, suggesting that HO-1 plays an important role in the suppression of NO production by EDG. Taken together, these results indicate that EDG isolated from Galla Rhois suppresses LPS-stimulated NO production in RAW 264.7 macrophages via HO-1 induction.

20S-dihydroprotopanaxadiol, a ginsenoside derivative, boosts innate immune responses of monocytes and macrophages

“…RAW264.7 (5×10 4 ) cells treated with 2H-PPD (0 to 40 μM) were resuspended in 100 μL PBS containing 1% human AB serum and incubated with fluorescein isothiocyanate (FITC)-dextran (1 mg/mL) at 37℃ for 2 h. The incubations were stopped by adding 2 mL ice-cold phosphate-buffered saline (PBS) containing 1% human serum and 0.02% sodium azide. The cells were then washed three times with cold PBS-azide and analyzed on a FACScan flow cytometer, as reported previously [21].…”

“…After incubation, cells were then incubated with SNP (0.25 mM), an inducer of ROS production, at 37℃ for 2 h. Cells were incubated with 20 μM of the fluorescent probe DHR123 for 1 h at 37℃. The degree of fluorescence, corresponding to intracellular ROS, was determined using a FACScan flow cytometer (Beckton-Dikinson, San Jose, CA, USA), as reported previously [21].…”

In Vitroantioxidative and anti-inflammatory activities of the ethanol extract of eggplant (Solanum melongena)stalks in macrophage RAW 264.7 cells

“…The cells were treated with S. melongena stalk fractions, incubated for an hour, and stimulated with lipopolysaccharide (LPS) 100 ng/mL (Sigma-Aldrich). After additional 24-h incubation, culture supernatant was collected, and 100 μL of Griess reagent (0.1% naphthylethylenediamine and 1% sulfanilamide in 5% phosphoric acid solution) was added to 100 μL of each supernatant (Park et al, 2011). NO production was then measured at 540 nm and quantified based on a sodium nitrate (NaNO 3 ) standard reference curve.…”

Pathophysiological Role of S-Nitrosylation and Transnitrosylation Depending on S-Nitrosoglutathione Levels Regulated by S-Nitrosoglutathione Reductase

“…Nitric oxide (NO) is a free radical that is formed in numerous cell types, including endothelial, muscle, and neuronal cells. NO can play the role of a physiological or pathological effector depending on the target signaling pathway ( NamKoong and Kim, 2010 ; Park et al ., 2011 ; Eo et al ., 2013 ; Ryu et al ., 2015 ). There are three subtypes of NO synthases (NOSs) in mammalian organisms, namely neuronal NOS (nNOS) and endothelial NOS (eNOS), which are constitutive forms, and inductive NOS (iNOS).…”