Isolation of anti-inflammatory compounds from Sambucus ebulus leaves through in vitro activity-guided fractionation

• Published in: Planta Medica
• Main Authors: Atay, İ, İlter, AZ, Bağatur, Y, Telci, D, Gören, AC, Kırmızıbekmez, H, Yeşilada, E
• Format: Conference Proceeding
• Language: English
• Published: 25.11.2015
• ISSN: 0032-0943; 1439-0221
• DOI: 10.1055/s-0035-1565693

The in vitro anti-inflammatory effects of subextracts ( n -hexane, chloroform, ethyl acetate, n -butanol, remaining water) of the methanol extract of the leaves of Sambucus ebulus L. (Adoxaceae) were investigated for their inhibitory activities on the activation of Nuclear factor kappa B (NF-κB) on lipopolysaccharide induced Raw 264.7 cells. The n -hexane, chloroform and ethyl acetate subextracts inhibited NF-κB activation at 50, 100 and 100 µg/mL concentrations, respectively. Two flavonoid mixtures [quercetin-3- O -β-D-glucopyranoside, quercetin-3- O -β-D-galactopyranoside], two flavonoids [isorhamnetin-3- O -β-D-glucopyranoside ( 1 ), isorhamnetin-3- O -rutinoside ( 2 )] were isolated from ethyl acetate subextract. 10- O -acetylpatrinoside ( 3 ) and a new iridoid [Sambulin B ( 4 )] was obtained from chloroform and n -hexane subextracts respectively. Structures were elucidated by NMR and MS. The compounds exerted inhibitions between 30 – 80% on NF-κB. Flavonoids were applied to cells at 25, 50, 75 and 100 µg/mL concentrations. Sambulin B was applied at 6,25, 12,5, 25 and Sambulin A applied at 12,5, 25 and 50 µg/mL concentrations. The effects on nitric oxide (NO), prostaglandine E 2 (PGE 2 ), tumor necrosis factor (TNF α ) and interleukins (IL-1 α , IL-1 β , IL-2, IL-6) were investigated by Griess and ELISA. The effects on iNOS, COX-2 protein levels and phosphorylation levels of mitogen activated protein kinases and I kappa B alpha (IκB α ) were examined by Western Blotting. 1 , 2 , 3 and 4 inhibited NO productions (between 59 – 84%), iNOS levels were decreased. 2, 3 and 4 exerted inhibitions on PGE 2 between 39 – 84%, COX-2 protein levels were decreased. 1 and 2 prevented p38/IκB α phosphorylations while 3 inhibited JNK/p38. Compound 4 inhibited JNK phosphorylation. All compounds (except mixtures) inhibited TNF α more than 29% and only 4 inhibited IL-6. Acknowledgements: This study is supported by TUBITAK-SBAG (Project no: 110S197) research grant.