ABSTRACT
Scorzonera species are used in different folk medicines to combat many diseases, including the illnesses connected with inflammation. Previous experiments showed anti-inflammatory activity of Scorzonera extracts in vivo. S. latifolia, S. cana var. jacquiniana, S. tomentosa, S. mollis ssp. szowitsii, S. eriophora, S. incisa, S. cinerea, and S. parviflora extracts were, therefore, evaluated for their inhibitory activities of TNF-α and IL-1ß production, and NF-κB nuclear translocation in THP-1 macrophages. The HPLC analysis was carried out to elucidate and to compare the composition of these extracts. Major compounds of the tested extracts have been isolated using different chromatographic techniques and further tested for their inhibitory activities on TNF-α and IL-1ß production. Several extracts showed promising anti-inflammatory activity in these in vitro tests. Results of HPLC analysis revealed chlorogenic acid as a compound present in all tested extracts. Hyperoside, quercetin-3-O-ß-d-glucoside and rutin were also present in varying amount in some Scorzonera species analyzed. Furthermore, eight phenolics which were identified as quercetin-3-O-ß-d-glucoside (1), hyperoside (2), hydrangenol-8-O-glucoside (3), swertisin (4), 7-methylisoorientin (5), 4,5-O-dicaffeoyl-quinic acid (6), 3,5-di-O-caffeoyl-quinic acid (7), and chlorogenic acid (8) have been isolated as major phenolic compounds of the tested extracts and, together with eight terpenoids (9-16) previously obtained from different Scorzonera species, have been tested for the inhibition of TNF-α production, unfortunately with no activity comparable with standard.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Cytokines/metabolism , NF-kappa B/metabolism , Plant Extracts/analysis , Scorzonera/chemistry , Anti-Inflammatory Agents/chemistry , Cell Line , Gene Expression Regulation/drug effects , Humans , In Vitro Techniques , Macrophages/drug effects , Macrophages/immunology , Phenols/chemistry , Phenols/pharmacology , Plant Extracts/pharmacology , Signal Transduction/drug effects , TurkeyABSTRACT
Juglone, 5-hydroxy-1,4-naphthoquinone, is the plant secondary metabolite with allelopathic properties, which was isolated especially from the plant species belonging to family Juglandaceae A. Rich. ex Kunth (walnut family). The mechanism of phytotoxic action of juglone was investigated on lettuce seedlings Lactuca sativa L. var. capitata L. cv. Merkurion by determining its effect at different levels. We have found that juglone inhibits mitosis (mitotic index 8.5 ± 0.6% for control versus 2.2 ± 0.9% for 250 µM juglone), changes mitotic phase index with accumulation of the cells in prophase (56.5 ± 2.6% for control versus 85.3 ± 5.0% for 250 µM juglone), and decreases meristematic activity in lettuce root tips (51.07 ± 3.62% for control versus 5.27 ± 2.29% for 250 µM juglone). In addition, juglone induced creation of reactive oxygen species and changed levels of reactive nitrogen species. Amount of malondialdehyde, a product of lipid peroxidation, increased from 24.0 ± 4.0 ng g(-1) FW for control to 55.5 ± 5.4 ng g(-1) FW for 250 µM juglone. We observed also changes in cellular structure, especially changes in the morphology of endoplasmic reticulum. Reactive oxygen species induced damage of plasma membrane. All these changes resulted in the disruption of the mitochondrial membrane potential, increase in free intracellular calcium ions, and DNA fragmentation and programmed cell death that was revealed by two methods, TUNEL test and DNA electrophoresis. The portion of TUNEL-positive cells increase from 0.96 ± 0.5% for control to 7.66 ± 1.5% for 250 µM juglone. Results of the study indicate complex mechanism of phytotoxic effect of juglone in lettuce root tips and may indicate mechanism of allelopathic activity of this compound.
