Vascular Protective Effects of Malus toringoides (Rehd.) Hughes Extracts and their Mechanism in Diabetic Rats and HUVECs.

Malus toringoides (Rehd.) Hughes (Rosaceae) is used as a traditional folk medicine in the Tibet autonomous region of China to treat hypertension, hyperglycemia, and hyperlipidemia. However, few modern pharmacological data on the use of this plant against diabetic syndrome are available. In this study, we examined the vascular protection provided by a 70% ethanol extract of M. toringoides (EMT) in human umbilical vein endothelial cells (HUVECs) grown in high-glucose medium and in a high-fat diet/streptozotocin-induced rat diabetes model. EMT significantly suppressed the expression of cell adhesion molecules in both HUVECs and diabetic rats. EMT also inhibited activation of the CX3CL1/CX3CR1 axis and the nuclear factor kappa B (NF-κB) signaling pathway in vivo and in vitro. The results provide a significant information on the vasoprotective properties of M. toringoides that may contribute to the development and application of related herbal medicines.

Kuwanon T and Sanggenon a Isolated from Morus alba Exert Anti-Inflammatory Effects by Regulating NF-κB and HO-1/Nrf2 Signaling Pathways in BV2 and RAW264.7 Cells.

We previously investigated the methanolic extract of Morus alba bark and characterized 11 compounds from the extract: kuwanon G (1), kuwanon E (2), kuwanon T (3), sanggenon A (4), sanggenon M (5), sanggenol A (6), mulberofuran B (7), mulberofuran G (8), moracin M (9), moracin O (10), and norartocarpanone (11). Herein, we investigated the anti-inflammatory effects of these compounds on microglial cells (BV2) and macrophages (RAW264.7). Among them, 3 and 4 markedly inhibited the lipopolysaccharide (LPS)-induced production of nitric oxide in these cells, suggesting the anti-inflammatory properties of these two compounds. These compounds inhibited the production of prostaglandin E2, interleukin-6, and tumor necrosis factor-α, and the expression of inducible nitric oxide synthase and cyclooxygenase-2 following LPS stimulation. Pretreatment with 3 and 4 inhibited the activation of the nuclear factor kappa B signaling pathway in both cell types. The compounds also induced the expression of heme oxygenase (HO)-1 through the activation of nuclear factor erythroid 2-related factor 2. Suppressing the activity of HO-1 reversed the anti-inflammatory effects caused by pretreatment with 3 and 4, suggesting that the anti-inflammatory effects were regulated by HO-1. Taken together, 3 and 4 are potential candidates for developing therapeutic and preventive agents for inflammatory diseases.

Anti-Inflammatory Effects of Compounds from Cudrania tricuspidata in HaCaT Human Keratinocytes.

The root bark of Cudrania tricuspidata has been reported to have anti-sclerotic, anti-inflammatory, antioxidant, neuroprotective, hepatoprotective, and cytotoxic activities. In the present study, the effect of 16 compounds from C. tricuspidata on tumor necrosis factor-α+interferon-γ-treated HaCaT cells were investigated. Among these 16 compounds, 11 decreased IL-6 production and 15 decreased IL-8 production. The six most effective compounds, namely, steppogenin (2), cudraflavone C (6), macluraxanthone B (12), 1,6,7-trihydroxy-2-(1,1-dimethyl-2-propenyl)-3- methoxyxanthone (13), cudraflavanone B (4), and cudratricusxanthone L (14), were selected for further experiments. These six compounds decreased the expression levels of chemokines, such as regulated on activation, normal T cell expressed and secreted (RANTES) and thymus and activation-regulated chemokine (TARC), and downregulated the protein expression levels of intercellular adhesion molecule-1. Compounds 2, 6, 12, 4, and 14 inhibited nuclear factor-kappa B p65 translocation to the nucleus; however, compound 13 showed no significant effects. In addition, extracellular signal regulatory kinase-1/2 phosphorylation was only inhibited by compound 14, whereas p38 phosphorylation was inhibited by compounds 13 and 4. Taken together, the compounds from C. tricuspidata showed potential to be further developed as therapeutic agents to suppress inflammation in skin cells.

Anti-Neuroinflammatory and Anti-Inflammatory Activities of Phenylheptatriyne Isolated from the Flowers of Coreopsis lanceolata L. via NF-κB Inhibition and HO-1 Expression in BV2 and RAW264.7 Cells.

Aging is associated with immune disregulation and oxidative stress which lead to inflammation and neurodegenerative diseases. We have tried to identify the anti-neuroinflammatory and anti-inflammatory components of Coreopsis lanceolata L. The dried flowers of C. lanceolata were extracted with 70% EtOH, and the obtained extract was divided into CH2Cl2, EtOAc, n-BuOH, and H2O fractions. The CH2Cl2 fraction was separated using silica gel and C-18 column chromatography to yield phenylheptatriyne (1), 2'-hydroxy-3,4,4'-trimethoxychalcone (2), and 4',7-dimethoxyflavanone (3). Additionally, the EtOAc fraction was subjected to silica gel, C-18, and Sephadex LH-20 column chromatography to yield 8-methoxybutin (4) and leptosidin (5). All the compounds isolated from C. lanceolata inhibited the production of nitric oxide (NO) in LPS-induced BV2 and RAW264.7 cells. In addition, phenylheptatriyne and 4',7-dimethoxyflavanone reduced the secretion of inflammatory cytokines, tumor necrosis factor alpha (TNF-α), and interleukin (IL)-6. Among them, phenylheptatriyne was significantly downregulated in the expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2). Subsequently, phenylheptatriyne also effectively inhibited nuclear factor-kappa B (NF-κB) activation in LPS-stimulated BV2 and RAW264.7 cells. Based on these results, the anti-neuroinflammatory effect of phenylheptatriyne isolated from C. lanceolata was confirmed, which may exert a therapeutic effect in treatment of neuroinflammation-related diseases.

The Anti-Oxidative and Anti-Neuroinflammatory Effects of Sargassum horneri by Heme Oxygenase-1 Induction in BV2 and HT22 Cells.

Sargassum horneri is used as a traditional medicinal agent and exhibits various pharmacological effects. In this study, we found that the 70% EtOH extract contained 34.37 ± 0.75 µg/mg fucosterol. We tested the antioxidant activities of the 70% EtOH extracts and their fractions. The CH2Cl2-soluble fraction showed the strongest DPPH and ABTS radical scavenging activities. Next, we evaluated the anti-neuroinflammatory effects of S. horneri on lipopolysaccharide (LPS)-stimulated BV2 cells. Pretreatment with the extract and fractions suppressed LPS-induced production of nitric oxide (NO) in BV2 cells. The 70% EtOH, CH2Cl2-soluble fraction, and water-soluble fraction inhibited the production of prostaglandin E2, interleukin-6, and tumor necrosis factor-α, as well as markedly blocking LPS-induced expression of inducible NO synthase and cyclooxygenase-2 via inactivation of the nuclear factor-kappa B pathway. In addition, the CH2Cl2-soluble fraction showed the most remarkable heme oxygenase (HO)-1 expression effects and increased nuclear erythroid 2-related factor translocation in the nucleus. In HT22 cells, the CH2Cl2-soluble fraction inhibited cell damage and ROS production caused by glutamate via the regulation of HO-1. Therefore, CH2Cl2-soluble fractions of S. horneri can attenuate oxidative action and neuroinflammatory responses via HO-1 induction, demonstrating their potential in the treatment of neuroinflammatory diseases.

Cudraflavanone B Isolated from the Root Bark of Cudrania tricuspidata Alleviates Lipopolysaccharide-Induced Inflammatory Responses by Downregulating NF-κB and ERK MAPK Signaling Pathways in RAW264.7 Macrophages and BV2 Microglia.

A prenylated flavonoid, cudraflavanone B, is isolated from Cudrania tricuspidata. In this study, we investigated its anti-inflammatory and anti-neuroinflammatory effects in lipopolysaccharide (LPS)-induced RAW264.7 and BV2 cells. In our initial study of the anti-inflammatory effects of cudraflavanone B the production of nitric oxide and prostaglandin E2 was attenuated in LPS-stimulated RAW264.7 and BV2 cells. These inhibitory effects were related to the downregulation of inducible nitric oxide synthase and cyclooxygenase-2. In addition, cudraflavanone B suppressed the production of pro-inflammatory cytokines such as interleukin-6 and tumor necrosis factor-α in LPS-induced RAW264.7 and BV2 cells. Moreover, the evaluation of the molecular mechanisms underlying the anti-inflammatory effects of cudraflavanone B revealed that the compound attenuated the nuclear factor-kappa B signaling pathway in LPS-induced RAW264.7 and BV2 cells. In addition, cudraflavanone B inhibited the phosphorylation of extracellular signal-regulated kinase mitogen-activated protein kinase signaling pathways in these LPS-stimulated cells. Thus, cudraflavanone B suppressed nuclear factor-κB, and extracellular signal-regulated kinase mitogen-activated protein kinase mediated inflammatory pathways, demonstrating its potential in the treatment of neuroinflammatory conditions.

Apigenin promotes antibacterial activity via regulation of nitric oxide and superoxide anion production.

Apigenin is a naturally occurring flavone isolated from the medicinal herb, Aster yomena. The present study was designed to elucidate the apoptosis-like antibacterial mechanism of apigenin in Escherichia coli. Administration of apigenin resulted in a rapid increase in intracellular calcium accompanied by an increase in reactive nitrogen species (RNS) and nitric oxide (NO) levels. Furthermore, apigenin increases reactive oxygen species (ROS), superoxide anion (O2 - ) which left E. coli with no ability to activate superoxide dismutase. Finally, we found that perturbance of the membrane lipid bilayer leading to glutathione oxidation and formation 8-hydroxy-2'-deoxyguanosine occurred during the process and apoptosis-like death hallmarks were further observed. Furthermore, we applied the NO synthase inhibitor ( l-NAME) and the O2 - scavenger (Tiron) and observed attenuation in apoptotic markers under their presence. Taken together, these results suggest that apigenin induces bacterial apoptosis via activation of cellular oxidative pathways dependent on the production and accumulation of RNS/ROS.

Neuroprotective and Anti-Inflammatory Effects of Kuwanon C from Cudrania tricuspidata Are Mediated by Heme Oxygenase-1 in HT22 Hippocampal Cells, RAW264.7 Macrophage, and BV2 Microglia.

Heme oxygenase (HO)-1 is a detoxifying phase II enzyme that plays a role in both inflammatory and oxidative stress responses. Curdrania tricuspidata is widespread throughout East Asia and is used as a therapeutic agent in traditional medicine. We investigated whether treatment with sixteen flavonoid or xanthone compounds from C. tricuspidata could induce HO-1 expression in HT22 hippocampal cells, RAW264.7 macrophage, and BV2 microglia. In these compounds, kuwanon C showed the most remarkable HO-1 expression effects. In addition, treatment with kuwanon C reduced cytoplasmic nuclear erythroid 2-related factor (Nrf2) expression and increased Nrf2 expression in the nucleus. Significant inhibition of glutamate-induced oxidative injury and induction of reactive oxygen species (ROS) occurred when HT22 hippocampal cells were pretreated with kuwanon C. The levels of inflammatory mediator and cytokine, which increased following lipopolysaccharide (LPS) stimulation, were suppressed in RAW264.7 macrophage and BV2 microglia after kuwanon C pretreatment. Kuwanon C also attenuated p65 DNA binding and translocation into the nucleus in LPS-induced RAW264.7 and BV2 cells. The anti-inflammatory, anti-neuroinflammatory, and neuroprotective effects of kuwanon C were reversed when co-treatment with HO-1 inhibitor of tin protoporphyrin-IX (SnPP). These results suggest that the neuroprotective and anti-inflammatory effects of kuwanon C are regulated by HO-1 expression.

Brassicaphenanthrene A from Brassica rapa protects HT22 neuronal cells through the regulation of Nrf2­mediated heme oxygenase­1 expression.

Brain cell damage that results from oxidative toxicity contributes to neuronal degeneration. The transcription factor nuclear factor­E2­related factor 2 (Nrf2) regulates the expression of heme oxygenase (HO)­1 and glutathione (GSH), and serves a key role in the pathogenesis of neurological diseases. Brassica rapa is a turnip that is unique to Ganghwa County, and is used mainly for making kimchi, a traditional Korean food. In the current study, brassicaphenanthrene A (BrPA) from B. rapa was demonstrated to exhibit protective effects against neurotoxicity induced by glutamate via Nrf2­mediated HO­1 expression. Similarly, BrPA increased the expression of cellular glutathione and glutamine­cysteine ligase genes. Furthermore, BrPA caused the nuclear translocation of Nrf2 and increased antioxidant response element (ARE) promoter activity. Nrf2 also mediated HO­1 induction by BrPA through the PI3K/Akt and JNK regulatory pathways. The results of the present study indicated the neuroprotective effect of BrPA, a natural food component from B. rapa.

Standardized microwave extract of Sappan Lignum exerts anti­inflammatory effects through inhibition of NF­κB activation via regulation of heme oxygenase­1 expression.

The extract of Sappan Lignum, the heartwood of Caesalpinia sappan L., has been used in medicine to improve blood circulation. Recently, the application of microwave extraction methods has been a major focus of research into the extraction of components from natural sources. In this experiment, we compared the anti­inflammatory effects of Sappan Lignum prepared by heat­70% EtOH extraction (CSE­H­70E) and microwave­70% EtOH extraction (CSE­MW­70E). High­performance liquid chromatography analysis was used to identify the compounds in these extracts. The heat­70% EtOH and microwave­70% EtOH extracts of Sappan Lignum had different chromatograms. CSE­MW­70E significantly inhibited the protein expression of iNOS and COX­2, PGE2, TNF­α, and reduced NO and IL­1ß production in macrophages exposed to LPS, whereas, only high concentrations of CSE­H­70E (20 µg/ml) resulted in any effects. Furthermore, CSE­MW­70E upregulated heme oxygenase­1 (HO­1) expression. In addition, the use of tin protoporphyrin, an inhibitor of HO­1, confirmed the inhibitory effects of CSE­MW­70E on pro­inflammatory mediators. These results suggested that the CSE­MW­70E­mediated upregulation of HO­1 played an important role in the anti­inflammatory effects of macrophages. Therefore, these findings showed that microwave extraction can be utilized to improve the extraction efficiency and biological activity of Sappan Lignum.

Synergistic Antifungal Activity of Isoquercitrin: Apoptosis and Membrane Permeabilization Related to Reactive Oxygen Species in Candida albicans.

Isoquercitrin (ISO), one of phytochemical isolated from aerial parts of Aster yomena, has been reported to have antifungal activity. However, the synergistic effect and the mechanism of ISO in combination with conventional antifungal agents are poorly understood. Therefore, synergistic antifungal effect between ISO and conventional antifungal agents was investigated. ISO at non-antifungal concentration interacts synergistically with amphotericin B (AMB) and fluconazole (FLC), but the combination with flucytosine (5-FC) showed no interaction. ISO disrupted an antioxidant system by inhibiting the activity of superoxide dismutase. This redox imbalance was shown to induce intracellular reactive oxygen species (ROS) accumulation and oxidative stress. ISO combined with FLC caused metacaspase activation and DNA condensation, markers of apoptosis, higher than the combination with ISO/AMB. In contrast, ISO with AMB synergistically stimulated membrane permeabilization compared to ISO/FLC. Scavenging ROS consequently reduced the synergy-induced apoptosis and membrane permeabilization, indicating combinations induced ROS were associated with the synergy effect of ISO. In conclusion, AMB and FLC enhanced the antifungal potency of ISO through oxidative stress when used in synergy. © 2018 IUBMB Life, 71(1):283-292, 2019.

Effect of apigenin isolated from Aster yomena against Candida albicans: apigenin-triggered apoptotic pathway regulated by mitochondrial calcium signaling.

ETHNOPHARMACOLOGICAL RELEVANCE: Aster yomena, a perennial herb that grows mainly in South Korea, has been employed in the traditional temple food for antibiotic efficacy. Recently, it was reported that apigenin isolated from A. yomena has a physical antifungal mechanism targeting membrane against Candida albicans. AIM OF THE STUDY: Our study aimed to investigate the biochemical responses underlying the antifungal activity of apigenin isolated from A. yomena due to lack studies reporting the investigation of intracellular responses of apigenin in C. albicans. MATERIALS AND METHODS: Apigenin was isolated from the aerial parts of A. yomena. To evaluate apigenin-induced inhibitory effects and membrane damages, the measurement of the cell viability assay and the flux of cytosolic components were performed with at various concentrations. Intracellular external potassium and calcium levels were assayed by an ion-selective electrode meter, Fura2-AM and Rhod2-AM, respectively. Mitochondrial dysfunctions were analyzed by using JC-1, Mitotracker Green FM, and MitoSOX Red dye. H2DCFDA, glutathione, and MDA assay were used to detect oxidative damage. Also, flow cytometry was carried out to detect apoptotic hallmarks using Annexin V-PI, TUNEL, and FITC-VAD-FMK staining. Tetraethylammoniumchloride (TEA), Ruthenium red (RR), and N-acetylcysteine (NAC) were used as a potassium channel blocker, mitochondrial calcium uptake inhibitor, and reactive oxygen species (ROS) scavenger, respectively. RESULTS: We confirmed that there was no decrease of cell survival percentages in crude extracts of A. yomena treatment, however, only isolated apigenin has the antifungal effect in C. albicans. Apigenin triggered a dose-dependent mitochondrial calcium uptake followed by mitochondrial dysfunction, loss of the membrane potential and an increase in the mitochondrial mass and ROS. Apigenin also induced intracellular redox imbalance as indicated by the ROS accumulation, glutathione oxidation, and lipid peroxidation. Interestingly, NAC failed the restore the mitochondrial calcium levels and thus alleviate the mitochondrial damages, however, RR reduced the apigenin-induced redox imbalance. Furthermore, apigenin induced apoptosis activation marked by the phosphatidylserine exposure, DNA fragmentation, and caspase activation. The pro-apoptotic effect of apigenin was counteracted by RR and NAC pretreatment. In particular, RR significantly reduced the pro-apoptotic responses. CONCLUSIONS: Apigenin isolated from A. yomena induced mitochondrial-mediated apoptotic pathway, and mitochondrial calcium signaling is main factor in its pathway in C. albicans.

Apigenin induces cell shrinkage in Candida albicans by membrane perturbation.

Apigenin, a natural flavone, has been well characterized for its their anticarcinogenic property; however, its bioactivity against pathogenic fungi has not been investigated in detail. In this study, we examined the antifungal activity and mode of action of apigenin. Apigenin inhibited the growth of fungal pathogens, which induced superficial infection and reduced biofilm mass. Three-dimensional flow cytometric analysis demonstrated that apigenin induced morphological changes, especially cell shrinkage, in Candida albicans. We investigated the cause of cell shrinkage using the cyanine dye 3,3΄-dipropylthiacarbocyanine iodide. Results revealed that apigenin altered the cell membrane potential. Apigenin also induced membrane dysfunction, and increased cell permeability to 1,6-diphenyl-1,3,5-hexatriene and propidium iodide. We observed the influx and efflux of fluorescent molecules of varying molecular weights and radii across large unilamellar vesicles and live cells that had been treated with apigenin. Membrane disruption facilitates the release of small intracellular constituents such as ions and sugars, but not proteins. These findings suggested that apigenin exerted an antifungal activity by inducing membrane disturbances, which led to cell shrinkage and leakage of intracellular components.

Effect of isoquercitrin on membrane dynamics and apoptosis-like death in Escherichia coli.

Minimum inhibitory concentration (MIC) is defined as the lowest concentration of a compound that completely inhibits microbial growth. Antibacterial mechanisms of compounds have been investigated at their sub-MICs as well as at their MIC. In this study, the effects of sub-MIC and MIC of isoquercitrin on Escherichia coli were investigated. The antibacterial effect of isoquercitrin was tested using the microdilution method. Sub-MICs of isoquercitrin induced the production of reactive oxygen species and depletion of glutathione. The oxidative effects induced by sub-MICs of isoquercitrin could be prolonged, finally resulting in apoptosis-like death. DNA fragmentation and phosphatidylserine externalization, which are regarded as the hallmarks of apoptosis, were evaluated using the TUNEL assay and Annexin V staining, respectively. Furthermore, isoquercitrin induced the peroxidation of membrane lipids and inner membrane permeabilization at both its sub-MIC and MIC. This suggested membrane damage in response to lipid oxidation. The uptake of membrane impermeable dyes, propidium iodide and calcein, demonstrated that isoquercitrin damaged the cell membrane at concentrations higher than its MIC. Thus, isoquercitrin induced apoptosis-like death and dysregulation of cell membrane dynamics.

Acerogenin C from Acer nikoense exhibits a neuroprotective effect in mouse hippocampal HT22 cell lines through the upregulation of Nrf-2/HO-1 signaling pathways.

Oxidative stress contributes to neuronal death in the brain, and neuronal death can cause aging or neurodegenerative disease. Heme oxygenase 1 (HO-1) serves a vital role in the regulation of biological reactions, including oxidative stress associated with reactive oxygen species. In the present study, acerogenin C isolated from the Aceraceae plant Acer nikoense, which is used as a Japanese folk medicine for hepatic disorders and eye diseases. However, there have been no studies on the mechanisms underlying the antineurodegenerative biological activities of acerogenin C. In the present study, acerogenin C demonstrated neuroprotective action against glutamate­induced cell death in hippocampal HT22 cells through the upregulation of HO­1 expression. These effects were also associated with nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) and the activation of phosphoinositide 3­kinase/protein kinase B. Taken together of the efficacy researches, this study determines that the Nrf2/HO­1 pathways denotes a biological mark and that acerogenin C might contribute to prevention of neurodegenerative disorders.

Oleanane triterpenoids from Akebiae Caulis exhibit inhibitory effects on Aß42 induced fibrillogenesis.

Previous phytochemical investigations of Akebiae Caulis resulted in the isolation of triterpenes, triterpene glycosides, phenylethanoid glycosides and megastigmane glycoside. Amyloid beta (Aß), the main component of the senile plaques detected in Alzheimer's disease, induces cell death. However, only a limited number of studies have addressed the biological and pharmacological effects of Akebiae Caulis. In particular, the inhibitory activity of Akebiae Caulis against Aß42 fibrillogenesis remains unclear. Herein, a new triterpene glycoside, akequintoside F (1), along with nine known compounds pulsatilla saponin A (2), collinsonidin (3), akebonic acid (4), hederagenin (5), 1-(3',4'-dihydroxycinnamoyl) cyclopentane-2,3-diol (6), asperosaponin C (7), leontoside A (8), quinatic acid (9), and quinatoside A (10) were isolated from Akebiae Caulis using repeated column chromatography with silica gel, LiChroprep RP-18, and MCI gel. The chemical structures of compounds 1-10 were illustrated based on 1D and 2D NMR spectroscopy, including 1H-1H COSY, HSQC, HMBC and NOESY spectroscopic analyses. Compound 1 a novel compound and known compounds 6 and 7 were isolated for the first time from this plant. Among these compounds, 1, 3, 4, 5 and 7 displayed significant inhibitory effects on Aß42 induced fibrillogenesis. We present the first report of new compound 1 and the inhibitory effects of components from Akebiae Caulis on Aß42 fibrillogenesis.

Phytol has antibacterial property by inducing oxidative stress response in Pseudomonas aeruginosa.

Phytol, isolated from Aster yomena, is widely distributed as a constituent of chlorophyll. In the present study, we confirmed the antibacterial activity of phytol and its mechanism inducing oxidative cell death in Pseudomonas aeruginosa. In phytol-treated cells, elevated level of intracellular reactive oxygen species (ROS) and transient NADH depletion were observed. These results demonstrated that phytol induced ROS accumulation and that the electron transport chain was involved in increase of ROS. Due to this ROS generation, the imbalance developed between intracellular ROS and the antioxidant defense system, leading to decrease of reduced glutathione (GSH). Moreover, severe DNA damage was shown after treatment with phytol. DNA electrophoresis and a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay were conducted with pretreatment with the antioxidant N-acetylcysteine (NAC) to evaluate the cause of DNA damage. In NAC-pretreated cells, alleviated damage was confirmed and it supports that phytol induces oxidative stress-mediated DNA damage. In conclusion, phytol exerts the antibacterial property via inducing oxidative stress response in P. aeruginosa.

Antibacterial Activity of Hibicuslide C on Multidrug-Resistant Pseudomonas aeruginosa Isolates.

Pseudomonas aeruginosa is a gram-negative bacterium that is frequently related to natural resistance to many drugs. In this work, the inhibition of growth against P. aeruginosa and multidrug-resistant P. aeruginosa (MDRPA) isolated from patients at Kyungpook National University was confirmed for hibicuslide C, essential oil components from Abutilon theophrasti. Hibicuslide C has antifungal activity with membrane disruption and apoptotic response against Candida albicans. However, its antibacterial activity was not reported yet. Cells treated with hibicuslide C was showed that its antipseudomonal activity is related to gDNA fragmentation and damage by TUNEL and gDNA electrophoresis. Furthermore, hibicuslide C worked synergistically with fluoroquinolones and rifampicin against MDRPA regardless of the ATP-associated mechanism. The antibiofilm activity possessed sole-resulting tissue culture plate method; besides that, the antibiofilm activity of other antibiotics was supported in particular MDRPA. The essential oil components like hibicuslide C may have antipseudomonal activity and, furthermore, increase in bacterial antibiotic susceptibility.

Moracin M inhibits airway inflammation by interrupting the JNK/c-Jun and NF-κB pathways in vitro and in vivo.

The therapeutic effectiveness of moracins as 2-arylbenzofuran derivatives against airway inflammation was examined. Moracin M, O, and R were isolated from the root barks of Morus alba, and they inhibited interleukin (IL)-6 production from IL-1ß-treated lung epithelial cells (A549) at 101-00µM. Among them, moracin M showed the strongest inhibitory effect (IC50=8.1µM). Downregulation of IL-6 expression by moracin M was mediated by interrupting the c-Jun N-terminal kinase (JNK)/c-Jun pathway. Moracin derivatives inhibited inducible nitric oxide synthase (iNOS)-catalyzed NO production from lipopolysaccharide (LPS)-treated alveolar macrophages (MH-S) at 50-100µM. In particular, moracin M inhibited NO production by downregulating iNOS. When orally administered, moracin M (20-60mg/kg) showed comparable inhibitory action with dexamethasone (30mg/kg) against LPS-induced lung inflammation, acute lung injury, in mice with that of dexamethasone (30mg/kg). The action mechanism included interfering with the activation of nuclear transcription factor-κB in inflamed lungs. Therefore, it is concluded that moracin M inhibited airway inflammation in vitro and in vivo, and it has therapeutic potential for treating lung inflammatory disorders.

A new indole glycoside from the seeds of Raphanus sativus.

A new indole glycoside, ß-D-glucopyranosyl 2-(methylthio)-1H-indole-3-carboxylate, named raphanuside A (1), as well as eight known compounds, ß-D-fructofuranosyl-(2 â†’ 1)-(6-O-sinapoyl)-α-D-glucopyranoside (2), (3-O-sinapoyl)-ß-D-fructofuranosyl-(2 â†’ 1)-α-D-glucopyranoside (3), (3-O-sinapoyl)-ß-D-fructofuranosyl-(2 â†’ 1)-(6-O-sinapoyl)-α-D-glucopyranoside (4), (3,4-O-disinapoyl)-ß-D-fructofuranosyl-(2 â†’ 1)-(6-O-sinapoyl)-α-D-glucopyranoside (5), isorhamnetin 3,4'-di-O-ß-D-glucoside (6), isorhamnetin 3-O-ß-D-glucoside-7-O-α-L-rhamnoside (7), isorhamnetin 3-O-ß-D-glucoside (8) and 3'-O-methyl-(-)-epicatechin 7-O-ß-D-glucoside (9) were isolated from the seeds of Raphanus sativus. Furthermore, compounds 1-3 and 6-9, were isolated from this plant for the first time. The structures of compounds 1-9 were identified using 1D and 2D NMR, including (1)H-(1)H COSY, HSQC, HMBC and NOESY spectroscopic analyses. The inhibitory activity of these isolated compounds against interleukin-6 (IL-6) production in TNF-α stimulated MG-63 cells was also examined.