Inhibition Of Nitric Oxide Production Research Articles

Metabolites Profile of Extracts and Fractions of Erythroxylum mexicanum Kunth by UHPLC-QTOF-MS/MS and its Antibacterial, Cytotoxic and Nitric Oxide Inhibitory Activities.

The present study shows the untargeted metabolite profiling and in vitro antibacterial, cytotoxic, and nitric oxide (NO) inhibitory activities of the methanolic leaves extract (MLE) and methanolic stem extract (MSE) of Erythroxylum mexicanum, as well as the fractions from MSE. Using ultra-high performance liquid chromatography/quadrupole time-of-flight tandem mass spectrometry (UHPLC-QTOF-MS/MS), a total of 70 metabolites were identified; mainly alkaloids in the MLE, while the MSE showed a high abundance of diterpenoids. The MSE fractions exhibited differential activity against Gram-positive bacteria. Notably, the hexane fraction (HSF) against Streptococcus pyogenes ATCC 19615 (MIC=62.5 μg/mL) exhibited a bactericidal effect. The MSE fractions exhibited cytotoxicity against all cancer cell lines tested, with selectivity towards them compared to a noncancerous cell line. Particularly, the HSF and chloroform fraction (CSF) showed the highest cytotoxicity against prostate cancer (PC-3) cells, with IC50 values of 19.9 and 18.1 μg/mL and selectivity indexes of 3.8 and 4.2, respectively. Both the HSF and ethyl acetate (EASF) fractions of the MSE inhibited NO production in RAW 264.7 macrophages, with NO production percentages of 50.0 % and 51.7 %, respectively, at a concentration of 30 μg/mL. These results indicated that E. mexicanum can be a source of antibacterial, cytotoxic, and anti-inflammatory metabolites.

Alkaloids and Lignans from the Aerial Parts of Rauvolfia tetraphylla Inhibit NO Production in LPS-activated RAW 264.7 Cells.

Three previously undescribed compounds named rauvolphyllas A-C (1-3), along with thirteen known compounds, 18β-hydroxy-3-epi-α-yohimbine (4), yohimbine (5), α-yohimbine (6), 17-epi-α-yohimbine (7), (E)-vallesiachotamine (8), (Z)-vallesiachotamine (9), 16S-E-isositsirikine (10), Nb -methylisoajimaline (11), Nb -methylajimaline (12), ajimaline (13), (+)-lyoniresinol 3α-O-β-D-glucopyranoside (14), (+)-isolarisiresinol 3α-O-β-D-glucopyranoside (15), and (-)-lyoniresinol 3α-O-β-D-glucopyranoside (16) were isolated from the aerial parts of Rauvolfia tetraphylla L. Their chemical structures were elucidated based on the extensive spectroscopic interpretation of HR-ESI-MS, 1D and 2D NMR spectra. The absolute configurations of 2 and 3 were determined by experimental ECD spectra. Compounds 5, 6, 7, and 11-13 exhibited nitric oxide production inhibition activity in LPS-activated RAW 264.7 cells with the IC50 values of 79.10, 44.34, 51.28, 33.54, 37.67, and 28.56 μM, respectively, compared to that of the positive control, dexamethasone, which showed IC50 value of 13.66 μM. The other isolates were inactive with IC50 values over 100 μM.

Anti-Herpes Simplex Virus and Anti-Inflammatory Activities of the Melittin Peptides Derived from Apis mellifera and Apis florea Venom.

Herpes simplex virus (HSV) is known to cause cold sores and various diseases in humans. Importantly, HSV infection can develop latent and recurrent infections, and it is also known to cause inflammation. These infections are difficult to control, and effective treatment of the disease remains a challenge. Thus, the search for new antiviral and anti-inflammatory agents is a necessity. Melittin is a major peptide that is present in the venom of the honeybee. It possesses a number of pharmacological properties. In this study, the effects of the melittin peptides from A. mellifera (MEL-AM) and A. florea (MEL-AF) against HSV-1 and HSV-2 were evaluated at different stages during the viral multiplication cycle in an attempt to define the mode of antiviral action using plaque reduction and virucidal assays. The results revealed a new finding that melittin at 5 µg/mL demonstrated the highest inhibitory effect on HSV through the direct inactivation of viral particles, and MEL-AF displayed a greater virucidal activity. Moreover, melittin was also observed to interfere with the process of HSV attachment to the host cells. MEL-AM exhibited anti-HSV-1 and anti-HSV-2 effects with EC50 values of 4.90 ± 0.15 and 4.39 ± 0.20 µg/mL, while MEL-AF demonstrated EC50 values of 4.47 ± 0.21 and 3.95 ± 0.61 µg/mL against HSV-1 and HSV-2, respectively. However, non-cytotoxic concentrations of both types of melittin produced only slight degrees of HSV-1 and HSV-2 inhibition after viral attachment, but melittin at 5 µg/mL was able to reduce the plaque size of HSV-2 when compared to the untreated group. In addition, MEL-AM and MEL-AF also exhibited anti-inflammatory activity via the inhibition of nitric oxide production in LPS-stimulated RAW 264.7 macrophage cells, and they were also found to down-regulate the expressions of the iNOS, COX-2 and IL-6 genes. The highest inhibition of IL-6 mRNA expression was found after treatment with 10 µg/mL of MEL-AM and MEL-AF. Therefore, melittin peptides have displayed strong potential to be used as an alternative treatment for HSV infection and inflammatory diseases in the future.

Metabolites of Culinary-medicinal Mushroom Hericium erinaceus Showed Anti-neuroinflammatory Activity in BV2 Cells and Anti-H2O2-induced Oxidative Stress Activity in SH-SY5Y Cells

Background: Hericium erinaceus is a versatile mushroom with potential therapeutic benefits. A variety of bioactive compounds have been found in this mushroom that have anti-ulcer, antiinflammatory, immunomodulatory, and blood sugar and lipid regulating effects. Objectives: The aim is to isolate and characterize compounds from the rice medium of H. erinaceus and evaluate their potential therapeutic effects on oxidative stress-associated diseases. Methods: Fifteen compounds were isolated and characterized using NMR spectra and ESI-MS. The effects of compounds 3 and 7 on LPS-induced nitric oxide (NO) generation in BV2 cells and H2O2- induced cytotoxicity in SH-SY5Y cells were investigated. The regulation of nitric oxide production and apoptosis-associated proteins Bcl-2 and Bax were also examined. Results: Compounds 3 and 7 exhibited potent inhibitory effects on LPS-induced NO generation in microglial cells and significant resistance to H2O2-induced cytotoxicity in neuronal cells. Molecular docking simulations demonstrated the interaction of compounds 3 and 7 with iNOS and Bax, respectively. Western blot analysis revealed that 7 inhibited NO production and regulated the expression of Bax and Bcl-2, suggesting a protective effect against H2O2-induced apoptosis. Conclusion: Compounds 3 and 7 isolated from H. erinaceus are potential therapeutic candidates for oxidative stress-associated diseases, possibly through their regulation of nitric oxide production and apoptosis-related proteins. Further investigations are warranted to elucidate their mechanisms of action and therapeutic potential.

Four New Steroidal Saponins from the Roots of Dracaena cambodiana with NO Production Inhibition Activity in LPS Activated RAW 264.7 Cells.

Seven steroidal saponins including three new 16,23-cyclocholestanes (1-3) and one new pregane (4) were isolated from the roots of Dracaena cambodiana Pierre ex Gagnep. Their chemical structures were elucidated to be (23R,25R)-26-O-β-D-glucopyranosyl-16,23-cyclocholesta-5,17(20)-dien-22-one-3β,16α,26-triol-3-O-α-L-rhamnopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→3)]-β-D-glucopyranoside (1), (23R,25R)-26-O-β-D-glucopyranosyl-16,23-cyclocholesta-5,17,20(22)-trien-3β,22,26-triol-3-O-α-L-rhamnopyranosyl-(1→3)-β-D-glucopyranoside (2), (23R,25R)-16,23-cyclocholesta-5,16,20(22)-trien-3β,22,26-triol-3-O-α-L-rhamnopyranosyl-(1→3)-β-D-glucopyranoside (3), 3β-[(O-α-L-rhamnopyranosyl-(1→3)-[α-L-rhamnopyranosyl-(1→2)]-β-D-gluco-pyranosyl)oxy]-pregna-5,17(20)-diene-16-one-20-carboxylic acid 4''''-O-β-D-glucopyranosylisopentyl ester (4), cambodianoside A (5), diosbulbiside C (6), and diosbulbiside D (7), by IR, HR-ESI-MS, 1D and 2D NMR spectra. Compounds 1 and 4-7 inhibited nitric oxide (NO) production in lipopolysaccharide activated RAW 264.7 cells with IC50 values ranging from 19.03±1.84 to 67.92±3.81 μM, whereas compounds 2 and 3 were inactive with IC50 values over 100 μM.

Nitric oxide production inhibitors from Polygonum multiflorum

This study focuses on the anti-inflammatory activity-guided fractionation of the root of Vietnamese Polygonum multiflorum, resulting in the isolation of ten compounds (1–10). These compounds were identified as trans-resveratrol (1), methyl protocatechuate (2), methyl gallate (3), catechin (4), epicatechin (5), 4,6-dihydroxy-2-O-(β- D-glucopyranosyl)acetophenone (6), quercetin (7), apigenin (8), luteolin (9), and tricin (10). Compounds 2, 3, and 6 were isolated from P. multiflorum for the first time. All isolated compounds were tested against lipopolysaccharide (LPS)-induced nitric oxide (NO) production in macrophage RAW264.7 cells to assess their anti-inflammatory potential. In vitro results revealed that compounds 7−9 exhibited significant NO production inhibition, with IC50 values of 12.0 ± 0.8, 17.8 ± 0.6, and 7.6 ± 0.3 μM, respectively. N(G)-monomethyl L-arginine (L-NMMA), a positive inhibitor, also effectively reduced LPS-induced NO production, with an IC50 value of 22.1 ± 0.1 μM. These findings indicate that compounds 7−9 isolated from P. multiflorum hold promise for further research and development of anti-inflammatory agents

Streptococcus mutans와 Porphyromonas gingivalis에 대한 상백피(Morus alba root bark) 에탄올 추출물의 항균 효과

This study aimed to analyze the antibacterial activity of the ethanol extract of Morus alba root bark (MAEE) against two critical oral pathogens, Streptococcus mutans and Porphyromonas gingivalis. The Griess reaction and WST-1 assays were used to analyze the nitric oxide (NO) inhibitory and cytotoxic effects of MAEE on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. The antibacterial activity of MAEE was evaluated using a disk diffusion assay and a growth inhibition assay by determining minimum inhibition concentrations (MICs) and minimum bactericidal concentrations (MBCs). MAEE treatment dose-dependently inhibited NO production and had no cytotoxic effect on LPS-stimulated RAW 264.7 cells. Furthermore, MAEE exhibited potent antibacterial activity and more potently generated a clear zone in the presence of S. mutans than P. gingivalis. The MICs of MAEE for S.mutans and P. gingivalis were 0.4 and 0.4~0.8 mg/mL, and its MBCs were 0.4 and 0.8 mg/mL, respectively. In addition, MAEE significantly inhibited the growths of both pathogens, but no dose-dependency was observed. The OD600 of S. mutans treated with MAEE at 3.2 mg/mL for 24 h was 0.146 compared to a control OD600 of 0.509. Absorbance at 600 nm for P. gingivalis treated with MAEE at 3.2 mg/mL for 24 h was 0.154 compared to a control OD600 of 0.486. In conclusion, MAEE showed potent antibacterial activity against both oral pathogens but affected S.mutans stronger than P. gingivalis. The study indicates that MAEE might help prevent dental caries and periodontal diseases.

Poly (vinyl alcohol)-gelatin-sericin copolymerized film fortified with vesicle-entrapped demethoxycurcumin/bisdemethoxycurcumin for improved stability, antibacterial, anti-inflammatory, and skin tissue regeneration

Vesicle delivery carriers, used to stabilize hydrophobic drugs, are characterized by the propensity to aggregate, and fuse, limiting its applications. Fortifying vesicle-entrapped drugs within a biodegradable polymeric film constitutes a promising solution. In this study, biodegradable poly (vinyl alcohol) copolymerized with gelatin-sericin film and integrated alongside vesicle-entrapped demethoxycurcumin (DMC) or bisdemethoxycurcumin (BDMC) was developed, extensively characterized for improve efficacy, and compared. Vesicle-entrapped DMC or BDMC was spherical in shape with no changes in size, zeta-potential, and morphology after storing at 4 °C for 30 days. Antibacterial activity of vesicle-entrapped DMC formulations against Acinetobacter baumannii and Staphylococcus epidermidis was more effective than that of its free form. DMC and BDMC demonstrated dose dependent reduction in lipopolysaccharides (LPS)-induced nitric oxide (NO) levels either in free or in entrapped form. Moreover, vesicle-entrapped DMC/BDMC suppressed NO production at lower concentrations, compared with that of their free form and significantly improved the viability of RAW264.7 and HaCaT cells. Furthermore, functionalized film with vesicle-entrapped DMC/BDMC demonstrated excellent radical scavenging, biocompatibility, and cell migration efficacy. Thus, incorporating vesicle, entrapped DMC/BDMC within biodegradable polymeric film may comprised a promising strategy for improving stability, wound healing, and inflammation attenuation efficacy.

3-Hydroxy-3-(2-oxopropyl)indolin-2-one, a product of a human-derived Enterocloster strain, is an inhibitor of nitric oxide production.

When cultured anaerobically, Enterocloster sp. RD014215 was found to produce 1. Using nuclear magnetic resonance and mass spectroscopy, the planar structure of 1 was determined to be 3-hydroxy-3-(2-oxopropyl)indolin-2-one. The chirality of 1 was implied as S by comparing the optical rotation value of 1 with literature reports of the synthesized compounds. To our knowledge, this work represents the first discovery of the metabolite produced by Enterocloster strain. 1 exhibited inhibition of nitric oxide (NO) production, demonstrating a 50% inhibitory activity (IC50) of 34 µm for NO production by murine macrophage cells subjected to lipopolysaccharide stimulation.

Polar Lipids of Marine Microalgae Nannochloropsis oceanica and Chlorococcum amblystomatis Mitigate the LPS-Induced Pro-Inflammatory Response in Macrophages.

Microalgae are recognized as a relevant source of bioactive compounds. Among these bioactive products, lipids, mainly glycolipids, have been shown to present immunomodulatory properties with the potential to mitigate chronic inflammation. This study aimed to evaluate the anti-inflammatory effect of polar lipids isolated from Nannochloropsis oceanica and Chlorococcum amblystomatis. Three fractions enriched in (1) digalactosyldiacylglycerol (DGDG) and sulfoquinovosyldiacylglycerol (SQDG), (2) monogalactosyldiacylglycerol (MGDG), and (3) diacylglyceryl-trimethylhomoserine (DGTS) and phospholipids (PL) were obtained from the total lipid extracts (TE) of N. oceanica and C. amblystomatis, and their anti-inflammatory effect was assessed by analyzing their capacity to counteract nitric oxide (NO) production and transcription of pro-inflammatory genes Nos2, Ptgs2, Tnfa, and Il1b in lipopolysaccharide (LPS)-activated macrophages. For both microalgae, TE and Fractions 1 and 3 strongly inhibited NO production, although to different extents. A strong reduction in the LPS-induced transcription of Nos2, Ptgs2, Tnfa, and Il1b was observed for N. oceanica and C. amblystomatis lipids. The most active fractions were the DGTS-and-PL-enriched fraction from N. oceanica and the DGDG-and-SQDG-enriched fraction from C. amblystomatis. Our results reveal that microalgae lipids have strong anti-inflammatory capacity and may be explored as functional ingredients or nutraceuticals, offering a natural solution to tackle chronic inflammation-associated diseases.

Microcurrent Cloth-Assisted Transdermal Penetration and Follicular Ducts Escape of Curcumin-Loaded Micelles for Enhanced Wound Healing.

Larger nanoparticles of bioactive compounds deposit high concentrations in follicular ducts after skin penetration. In this study, we investigated the effects of microcurrent cloth on the skin penetration and translocation of large nanoparticle applied for wound repair applications. A self-assembly of curcumin-loaded micelles (CMs) was prepared to improve the water solubility and transdermal efficiency of curcumin. Microcurrent cloth (M) was produced by Zn/Ag electrofabric printing to facilitate iontophoretic transdermal delivery. The transdermal performance of CMs combined with M was evaluated by a transdermal system and confocal microscopy. The CMs/iontophoretic combination effects on nitric oxide (NO) production and inflammatory cytokines were evaluated in Raw 264.7 cells. The wound-healing property of the combined treatment was assessed in a surgically created full-thickness circular wound mouse model. Energy-dispersive X-ray spectroscopy confirmed the presence of Zn/Ag on the microcurrent cloth. The average potential of M was measured to be +214.6 mV in PBS. Large particle CMs (CM-L) prepared using surfactant/cosurfactant present a particle size of 142.9 nm with a polydispersity index of 0.319. The solubility of curcumin in CM-L was 2143.67 μg/mL, indicating 250-fold higher than native curcumin (8.68 μg/mL). The combined treatment (CM-L+M) demonstrated a significant ability to inhibit NO production and increase IL-6 and IL-10 secretion. Surprisingly, microcurrent application significantly improved 20.01-fold transdermal performance of curcumin in CM-L with an obvious escape of CM-L from follicular ducts to surrounding observed by confocal microscopy. The CM-L+M group also exhibited a better wound-closure rate (77.94% on day 4) and the regenerated collagen intensity was approximately 2.66-fold higher than the control group, with a closure rate greater than 90% on day 8 in vivo. Microcurrent cloth play as a promising iontophoretic transdermal drug delivery accelerator that enhances skin penetration and assists CMs to escape from follicular ducts for wound repair applications.

배, 맥아, 대추, 익모초, 황금 및 이들 혼합물의 생리활성

This study investigated the anti-inflammatory, trypsin activity, and antioxidant effects of 11 kinds of plant extracts to discover materials for developing optimal mixtures that improve inflammation and help digestion. Ziziphus jujuba Mill. (ZJ), Leonurus japonicus Houtt. (LJ), Scutellaria baicalensis (SB), Platycodon grandiflorum, and Aster scaber extracts had excellent anti-inflammatory effects by reducing excessive nitric oxide (NO) and tumor necrosis factor-α content in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. The malt (MA), Pyrus pyrifolia Nakai (PP), Raphanus sativus L., Platycodon grandiflorum extracts among the 11 kinds of plant extracts had high trypsin activity. The antioxidant activity of the plant extracts was examined by the DPPH radical scavenging activity, and the SB, PE, JU, and MA extracts had high antioxidant activity. Therefore, PP, MA, ZJ, LJ, and SB were selected to develop optimal mixtures that improve inflammation and help digestion. The extract of plant mixture containing PP, MA, ZJ, LJ, and SB in the ratio 1:1:2:1:2 (w/w) significantly inhibited NO production than the extract of PP, MA, ZJ, LJ, and SB, respectively, in LPS-stimulated RAW 264.7 macrophages. The DPPH radical scavenging activity of the mixture extract was significantly higher than the extract of PP, MA, ZJ, and LJ, respectively.

Optimization of curcumin extraction from Curcuma domestica Vahl. rhizome with microwave-assisted extraction technique in inhibiting nitric oxide production and cell viability

Curcumin is a yellow dye derived from the turmeric plant (Curcuma domestica Vahl.). In turmeric rhizome, there are active compounds that are often used as traditional medicine in the form of curcuminoids. The microwave-Assisted Extraction (MAE) technique is an extraction technique that utilizes microwave radiation to heat the solvent quickly and efficiently and is also very suitable for extracting compounds that are not resistant to heat. This research was conducted to determine the level of curcumin from C. domestica Vahl. Extract with MAE using virgin coconut oil (VCO) as solvent. Extraction of curcumin was carried out using C. domestica Vahl powder with various weights (1 g, 1.5 g, 2 g, and 2.5 g) and VCO as solvent (mL), with an extraction time of 10 mins and power of 270 W. Curcumin from the extract was measured using High-Performance Liquid Chromatography (HPLC). The results showed that the method has a limit of detection (LOD), which is 3.57±0.01 µg/mL, a limit of quantification (LOQ) of 11.91±0.04 ppm, and good linearity with a correlation coefficient (r) of 0.9993. The curcumin level was determined with HPLC from each various weight (1.27±0.03 mg/g); 1.5 g (1.20±0.02 mg/ g); 2 g (3.45±0.02 mg/g) and 2.5 g (3.44±0.01 mg/g) respectively. The results showed that the optimum weight for curcumin extraction with MAE is 2 g with 20 mL of VCO with 10 mins extraction time and power (270 W). The best results were shown at a 1.56 µg/mL concentration and had the highest % of living cells, cell viability testing showed that curcumin was not toxic to RAW 264.7 cells. The nitric oxide (NO) production results show that cells stimulated with LPS had higher nitrite levels at 31.25 g/mL and lower nitrite levels at 62.5 µg/mL; 125 µg/mL; 250 µg/mL, implying that curcumin MAE extract can inhibit NO production, whereas extract-stimulated cells showed a decrease in nitrite levels.

Inhibitory efficiency of Andrographis paniculata extract on viral multiplication and nitric oxide production.

Andrographis paniculata (Burm. F.) Nees is a medicinal plant previously reported with broad-spectrum antivirals but the mode of inhibition remains elusive. The objective of this study was to identify the most active fraction from A. paniculata ethanol extract (APE, APE-2A, APE-2B and APE-2C) and dry powder extract (APSP) against influenza A (H3N2), representing RNA viruses, and herpes simplex virus-1 (HSV-1), representing DNA viruses. The results showed that the fractions APSP, APE, APE-2B, and APE-2C directly neutralized the HSV-1 and influenza A (H3N2) when incubated at room temperature for 60min before infecting the cells. The results also showed that the additional APE-2A fraction also directly neutralized the influenza A (H3N2), but not the HSV-1. The APE, APE-2B and APE-2C inhibited the HSV-1 by more than 0.5 log when the fractions were introduced after infection. Similarly, the APSP and APE inhibited the influenza A (H3N2) more than 0.5 log after infection. Only 50μg/mL APE-2C inhibited the viruses greater than 0.5 log. In addition, A. paniculata extracts were also evaluated for their interfering capacities against nitric oxide (NO) production in LPS-activated RAW 264.7 macrophages. As well, APE-2C potently inhibited NO production at the IC50 of 6.08μg/mL. HPLC and LC-MS analysis indicated that the most actively antiviral fractions did not contain any andrographolide derivatives, whereas the andrographolide-rich fractions showed moderate activity.

α-Mangostin and lawsone methyl ether in tooth gel synergistically increase its antimicrobial and antibiofilm formation effects in vitro

Objectivesα-Mangostin (α-MG) and lawsone methyl ether (LME) show antimicrobial and anti-biofilm activities. The objectives of this study were to develop a herbal tooth gel containing α-MG and LME plus fluoride and determine its antimicrobial, anti-biofilm formation, anti-cancer, anti-inflammatory, wound healing, and enamel microhardness effects.MethodsAntimicrobial assays against Streptococcus mutans, Porphyromonas gingivalis, and Candida albicans were performed. The microbes’ ultrastructural morphology was assessed using Transmission Electron Microscopy. The effect on microbial biofilm formation was tested by a broth microdilution. Cell viability was assessed with MTT assay. The anti-inflammatory effect was investigated by measuring inhibition of nitric oxide production. Enamel microhardness was measured via Vickers microhardness testing. The enamel chemical composition was investigated with Fourier Transform Spectrometer. The enamel surface morphology and fluoride content were examined by Scanning Electron Microscopy and Energy Dispersive X-ray Spectroscopy.ResultsThe results show synergistic effects of α-MG and LME on antimicrobial activity and antibiofilm formation without cytotoxicity at a therapeutic dose. At a higher dose, the tooth gel inhibited proliferation of cancer cell line. Enamel microhardness was increased after brushing with the tooth gel plus fluoride. A large amount of fluoride was detected on the enamel surface.ConclusionThe tooth gel containing α-MG and LME synergized its antimicrobial activity and antibiofilm formation and inhibited oral cancer cell proliferation. Incorporating fluoride into the tooth gel increased enamel microhardness. Thus, the herbal tooth gel containing α-MG and LME plus fluoride may be useful for preventing dental caries and promoting oral health.

GC-MS Characterization, in Vitro Antioxidant and Anti-Inflammatory Activities of Essential oil from the Leaves of Litsea balansae Lecomte

Objective: This study aimed to analyze the essential oil obtained from Litsea balansae Lecomte leaves and investigate its potential as an anti-inflammatory and antioxidant agent. Methods: Fresh L balansae leaves were collected in Quangtri, Vietnam, and subjected to hydro-distillation using a Clevenger-type apparatus to extract the essential oil. The essential oil composition was analyzed using gas chromatography–mass spectrometry (GC-MS). Antioxidant capacity was evaluated through the 2,2-Diphenyl-1-picrylhydrazyl assay, while the inhibition of nitric oxide production was assessed in lipopolysaccharide-activated RAW264.7 cells. Results: GC-MS analysis identified 71 compounds, constituting 99.3% of the essential oil. Key constituents included carvacrol (20.8%), caryophyllene oxide (11.0%), cis-β-guaiene (6.4%), khusimone (3.9%), bicycloelemene (3.7%), and spathulenol (3.7%). The essential oil exhibited a moderate antioxidant effect (half maximal inhibitory concentration [IC50] = 208.37 ± 10.67 µg/mL) and significant anti-inflammatory activity (IC50 = 28.09 ± 1.53 µg/mL). Conclusion: The essential oil derived from L balansae leaves showcases potential as an anti-inflammatory and antioxidant agent, as indicated by its chemical composition and biological activities. These findings suggest promising avenues for further research in the development of therapeutic agents from natural sources.

Zebrafish nos2a benefits bacterial proliferation via suppressing ROS and inducing NO production to impair the expressions of inflammatory cytokines and antibacterial genes

The enzyme nitric oxide synthase 2 or inducible NOS (NOS2), reactive oxygen species (ROS) and nitric oxide (NO) are important participants in various inflammatory and immune responses. However, the functional significances of the correlations among piscine NOS2, ROS and NO during pathogen infection remain unclear. In teleost, there are two nos2 genes (nos2a and nos2b). It has been previously reported that zebrafish nos2a behaves as a classical inducible NOS, and nos2b exerts some functions similar to mammalian NOS3. In the present study, we reported the functional characterization of zebrafish nos2a during bacterial infection. We found that zebrafish nos2a promoted bacterial proliferation, accompanied by an increased susceptibility to Edwardsiella piscicida infection. The nagative regulation of zebrafish nos2a during E. piscicida infection was characterized by the impaired ROS levels, the induced NO production and the decreased expressions of proinflammatory cytokines, antibacterial genes and oxidant factors. Furthermore, although both inducing ROS and inhibiting NO production significantly inhibited bacterial proliferation, only inhibiting NO production but not inducing ROS significantly increased resistance to E. piscicida infection. More importantly, ROS supplementation and inhibition of NO completely abolished this detrimental consequence mediated by zebrafish nos2a during E. piscicida infection. All together, these results firstly demonstrate that the innate response mediated by zebrafish nos2a in promoting bacterial proliferation is dependent on the lower ROS level and higher NO production. The present study also reveals that inhibition of NO can be effective in the protection against E. piscicida infection.

PolyQ-Expansion Causes Mitochondria Fragmentation Independent of Huntingtin and Is Distinct from Traumatic Brain Injury (TBI)/Mechanical Stress-Mediated Fragmentation Which Results from Cell Death.

Mitochondrial dysfunction has been reported in many Huntington's disease (HD) models; however, it is unclear how these defects occur. Here, we test the hypothesis that excess pathogenic huntingtin (HTT) impairs mitochondrial homeostasis, using Drosophila genetics and pharmacological inhibitors in HD and polyQ-expansion disease models and in a mechanical stress-induced traumatic brain injury (TBI) model. Expression of pathogenic HTT caused fragmented mitochondria compared to normal HTT, but HTT did not co-localize with mitochondria under normal or pathogenic conditions. Expression of pathogenic polyQ (127Q) alone or in the context of Machado Joseph Disease (MJD) caused fragmented mitochondria. While mitochondrial fragmentation was not dependent on the cellular location of polyQ accumulations, the expression of a chaperone protein, excess of mitofusin (MFN), or depletion of dynamin-related protein 1 (DRP1) rescued fragmentation. Intriguingly, a higher concentration of nitric oxide (NO) was observed in polyQ-expressing larval brains and inhibiting NO production rescued polyQ-mediated fragmented mitochondria, postulating that DRP1 nitrosylation could contribute to excess fission. Furthermore, while excess PI3K, which suppresses polyQ-induced cell death, did not rescue polyQ-mediated fragmentation, it did rescue fragmentation caused by mechanical stress/TBI. Together, our observations suggest that pathogenic polyQ alone is sufficient to cause DRP1-dependent mitochondrial fragmentation upstream of cell death, uncovering distinct physiological mechanisms for mitochondrial dysfunction in polyQ disease and mechanical stress.

New Monoterpene Glycoside Paeoniflorin Derivatives as NO and IL-1β Inhibitors: Synthesis and Biological Evaluation.

Several monoterpene glycoside compounds were extracted from Paeonia lactiflora Pall. Among them, paeoniflorin, a water-soluble monoterpene glycoside found in the root of Paeonia lactiflora Pall, exhibits excellent antioxidant pharmacological functions. Initially, Sc(CF3SO3)3 was employed as the catalyst for paeoniflorin's dehydration and rearrangement reactions with alcohols. Subsequently, structural modifications were performed on paeoniflorin through a series of responses, including acetylation, deacetylation, and debenzoylation, ultimately yielding 46 monoterpene glycoside derivatives. The potential inhibitory effects on the pro-inflammatory mediators interleukin-1 beta (IL-1β) and nitric oxide (NO) were assessed in vitro. The results revealed that compounds 29 and 31 demonstrated notable inhibition of NO production, while eight derivatives (3, 8, 18, 20, 21, 29, 34, and 40) displayed substantial inhibitory effects on the secretion of IL-1β. Computational research was also undertaken to investigate the binding affinity of the ligands with the target proteins. Interactions between the proteins and substrates were elucidated, and corresponding binding energies were calculated accordingly. The findings of this study could provide valuable insights into the design and development of novel anti-inflammatory agents with enhanced pharmacological properties.

Exosome-like nanoparticles derived from Allium tuberosum prevent neuroinflammation in microglia-like cells.

Exosome-like nanoparticles (ELNs), which are plant-derived extracellular membrane vesicles, can regulate mammalian gene expression. ELNs can cross the blood-brain barrier, making them potential therapeutic agents or drug-delivery carriers for neuroinflammation-related diseases. Here, we investigated the anti-neuroinflammatory potential of ELNs extracted from Allium tuberosum (A-ELNs). A-ELNs were extracted, and their miRNA profile was characterized. A-ELNs were also applied to BV-2 microglial and MG-6 cells derived from C57/BL6 mice stimulated with lipopolysaccharide (LPS), followed by an examination of levels of inflammatory-related factors. To test their drug-carrying potential, A-ELNs were mixed with dexamethasone, an anti-inflammatory drug, to prepare dexamethasone-incorporated A-ELNs (Dex-A-ELNs). A-ELNs showed a particle size of 145 ± 2 nm and characteristic miRNAs. A-ELNs significantly decreased the LPS-induced nitric oxide (NO) and inflammatory cytokines levels in BV-2 and MG-6 cells. The mRNA expression of heme oxygenase-1 was significantly increased, and that of inducible NO synthase and inflammatory cytokines was significantly decreased by A-ELNs in BV-2 cells. Dex-A-ELNs inhibited NO production in BV-2 cells more potently than either A-ELNs or dexamethasone alone. A-ELNs can alleviate microglial inflammation. Their effects can be potentiated by incorporating anti-inflammatory drugs, such as dexamethasone, making them potential therapeutic agents or drug-delivery carriers for neuroinflammation.