İrem Atay Balkan1, Ayca Zeynep İlter Akülke2, Yeşim Bağatur3, Dilek Telci4, Ahmet Ceyhan Gören5, Hasan Kırmızıbekmez6, Erdem Yesilada7. 1. Yeditepe University, Faculty of Pharmacy, Department of Pharmacognosy and Phytotherapy, 34755 Ataşehir, İstanbul, Turkey. Electronic address: irematay@yahoo.com. 2. Yeditepe University, Faculty of Engineering, Department of Genetics and Bioengineering, 34755 Ataşehir, İstanbul, Turkey. Electronic address: aycazeynep@gmail.com. 3. Yeditepe University, Faculty of Engineering, Department of Genetics and Bioengineering, 34755 Ataşehir, İstanbul, Turkey. Electronic address: yesimbagatur@gmail.com. 4. Yeditepe University, Faculty of Engineering, Department of Genetics and Bioengineering, 34755 Ataşehir, İstanbul, Turkey. Electronic address: dtelci@gmail.com. 5. TUBITAK National Metrology Institute, Chemistry Group Laboratories, 41470 Gebze, Kocaeli, Turkey. Electronic address: ahmetceyhan.goren@tubitak.gov.tr. 6. Yeditepe University, Faculty of Pharmacy, Department of Pharmacognosy and Phytotherapy, 34755 Ataşehir, İstanbul, Turkey. 7. Yeditepe University, Faculty of Pharmacy, Department of Pharmacognosy and Phytotherapy, 34755 Ataşehir, İstanbul, Turkey. Electronic address: yesilada@yeditepe.edu.tr.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: The leaves of Sambucus ebulus L. (Adoxaceae) are widely used in Turkish folk medicine particularly against inflammatory disorders. The fresh leaves after wilted over fire or the poultices prepared are directly applied externally to heal burns, edema, eczema, urticarial and abscess. Two iridoids were recently isolated (sambulin A, sambulin B) from the leaves of S. ebulus. AIM OF THE STUDY: This study aims to investigate the in vitro anti-inflammatory activities of these iridoids on LPS-induced RAW 264.7 macrophages. MATERIALS AND METHODS: Raw 264.7 macrophages were treated with 12.5, 25 and 50µg/ml Sambulin A and 6.25, 12.5 and 25µg/ml Sambulin B and induced with 1µg/ml lipopolysaccaharides (LPS). Effect of the compounds on nitric oxide (NO) production and cytokines (TNFα, IL-6) were determined by Griess and ELISA assays respectively. iNOS and the phosphorylation levels of MAPKs (ERK, JNK) were examined by Western Blot. RESULTS: Sambulin A and sambulin B inhibited 52.82% and 72.88% of NO production at 50 and 25µg/ml concentrations respectively. The levels of iNOS were significantly decreased by both molecules, sambulin B at 25µg/ml almost completely decreased iNOS levels (97.53%). Both molecules significantly inhibited TNFα productions. However, only sambulin B inhibited IL-6 production. Consequently, it was shown that sambulin B exerted its effect through the inhibition of ERK and JNK phosphorylations. CONCLUSION: The prominent bioactivities exerted by two iridoids will contribute to explanation of the usage of S. ebulus in traditional medicine against rheumatoid diseases.
ETHNOPHARMACOLOGICAL RELEVANCE: The leaves of Sambucus ebulus L. (Adoxaceae) are widely used in Turkish folk medicine particularly against inflammatory disorders. The fresh leaves after wilted over fire or the poultices prepared are directly applied externally to heal burns, edema, eczema, urticarial and abscess. Two iridoids were recently isolated (sambulin A, sambulin B) from the leaves of S. ebulus. AIM OF THE STUDY: This study aims to investigate the in vitro anti-inflammatory activities of these iridoids on LPS-induced RAW 264.7 macrophages. MATERIALS AND METHODS: Raw 264.7 macrophages were treated with 12.5, 25 and 50µg/ml Sambulin A and 6.25, 12.5 and 25µg/ml Sambulin B and induced with 1µg/ml lipopolysaccaharides (LPS). Effect of the compounds on nitric oxide (NO) production and cytokines (TNFα, IL-6) were determined by Griess and ELISA assays respectively. iNOS and the phosphorylation levels of MAPKs (ERK, JNK) were examined by Western Blot. RESULTS:Sambulin A and sambulin B inhibited 52.82% and 72.88% of NO production at 50 and 25µg/ml concentrations respectively. The levels of iNOS were significantly decreased by both molecules, sambulin B at 25µg/ml almost completely decreased iNOS levels (97.53%). Both molecules significantly inhibited TNFα productions. However, only sambulin B inhibited IL-6 production. Consequently, it was shown that sambulin B exerted its effect through the inhibition of ERK and JNK phosphorylations. CONCLUSION: The prominent bioactivities exerted by two iridoids will contribute to explanation of the usage of S. ebulus in traditional medicine against rheumatoid diseases.