Oxide Production In Lipopolysaccharide-stimulated RAW Research Articles

Spermacosides a and B, two new triterpenoid saponins from Spermacoce ocymoides burm.f.

Two new triterpenoid saponins, named spermacosides A–B (1 − 2), together with two known oleanane-type triterpenoid saponins, 3-O-β-D-xylopyranosyl-(1→3)-β-D-glucopyranosylbayogenin (3) and 3-O-β-D-glucopyranosylbayogenin (4), were isolated from the ethyl acetate extract of Spermacoce ocymoides Burm.f. in a phytochemical investigation. Their chemical structures were elucidated by spectroscopic data analysis (1D and 2D NMR, and HR-ESI-MS), as well as comparison with reported data. All these compounds were evaluated for inhibiting nitric oxide production in lipopolysaccharide-stimulated RAW 264.7 cells. Among them, 1 showed a slight effect with an IC50 value of 108.65 ± 7.91 µM, and compounds 2–4 were inactive.

Six unprecedented 2-(2-phenethyl)chromone dimers from agarwood of Aquilaria filaria

Six new dimeric 2-(2-phenylethyl)chromones (1–6) were successfully isolated from the ethanol extract of agarwood of Aquilaria filaria from Philippines under HPLC-MS guidance. Compounds 1–6 are all dimers formed by linking 5,6,7,8-tetrahydro-2-(2-phenylethyl)chromone and flindersia 2-(2-phenylethyl)chromone via a single ether bond, and the linkage site (C5-O-C8′′) of compound 2 is extremely rare. A variety of spectroscopic methods were used to ascertain their structures, including extensive 1D and 2D NMR spectroscopic analysis, HRESIMS, and comparison with literature. The in vitro tyrosinase inhibitory and anti-inflammatory activities of each isolate were assessed. Among these compounds, compound 2 had a tyrosinase inhibition effect with an IC50 value of 27.71 ± 2.60 μM, and compound 4 exhibited moderate inhibition of nitric oxide production in lipopolysaccharide-stimulated RAW264.7 cells with an IC50 value of 35.40 ± 1.04 μM.

Two unprecedented 2-(2-phenethyl)chromone dimers from red soil agarwood of Aquilaria crassna

Two unprecedented 2-(2-phenethyl)chromone dimers, aquicrassones A and B (1 and 2), were isolated by HPLC-MS guided method from the ethanol extract of red soil agarwood originating from Aquilaria crassna in Vietnam. Their structures were unambiguously elucidated by analysis of HR-ESI-MS, 1D and 2D NMR, CD data and by comparison with literature data. Compounds 1 and 2 were evaluated for their inhibition effect on nitric oxide production in lipopolysaccharide-stimulated RAW264.7 cells. Compound 2 displayed anti-inflammatory activity with an IC50 value of 20.36 ± 0.41 μM.

Phytochemical profile and anti-inflammatory activity of the hull of γ-irradiated wheat mutant lines (Triticum aestivum L.).

Wheat (Triticum aestivum Linn.; Poaceae) is the second most cultivated food crop among all global cereal crop production. The high carbohydrate content of its grains provides energy, multiple nutrients, and dietary fiber. After threshing, a substantial amount of wheat hull is produced, which serves as the non-food component of wheat. For the valorization of these by-products as a new resource from which functional components can be extracted, the hull from the seeds of cultivated wheat mutant lines bred after γ-irradiation were collected. Untargeted metabolite analysis of the hull of the original cultivar (a crossbreeding cultivar., Woori-mil × D-7) and its 983 mutant lines were conducted using ultra-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry technique. A total of 55 molecules were tentatively identified, including 21 compounds found in the Triticum species for the first time and 13 compounds not previously described. Among them, seven flavonolignans with a diastereomeric structure, isolated as a single compound from the hull of T. aestivum in our previous study, were used as the standards in the metabolite analysis. The differences in their collision cross-section values were shown to contribute to the clear distinction between tricine-lignan stereoisomers. To select functionally active agents with anti-inflammatory activity among the identified compounds, the wheat hull samples were evaluated for their inhibitory effect on nitric oxide production in lipopolysaccharide-stimulated RAW 264.7 cells. As a result of multivariate analysis based on the results of chemical and biological profiles of the wheat hull samples, 10 metabolites were identified as key markers, contributing to the distinction between active and inactive mutant lines. Considering that one of the four key markers attributed to anti-inflammatory activity has been identified to be a flavonolignan, the wheat hull could be a valuable source of diverse tricin-lignan type compounds and used as a natural health-promoting product in food supplements.

Effects of the drying method and extraction solvent on antioxidant and anti-inflammatory activity of Melosira nummuloides bioproducts

Melosira nummuloides is a marine diatom with potential use as food, fuel, and a dietary supplement. However, the efficacy of its extraction and drying techniques have not been explored. Here, M. nummuloides powders were prepared by two drying methods—hot-air drying (HAD) and freeze-drying (FD)—and extracted with hot water, ethanol, methanol, and chloroform:methanol (CM) at a ratio of 2:1 v/v. The antioxidant and anti-inflammatory activity of each extract was investigated. The CM extract had the greatest 2,2-diphenyl-1-picrylhydrazyl and 2,2ʹ-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity among the solvent extracts, and a slight difference in antioxidant activity was observed across the various drying methods. Compared to other extracts, both the FD-CM and HAD-CM extracts showed stronger anti-inflammatory effects by inhibiting nitric oxide production in lipopolysaccharide-stimulated RAW 264.7 cells. Furthermore, the FD-CM extract contained a wide range of lipophilic compounds. Notably, myristic acid (29.08 ± 0.45 mg/g dry weight powder extract (DW)), oleic acid (25.20 ± 0.92 mg/g DW), palmitoleic acid (10.77 ± 0.41 mg/g DW), eicosapentaenoic acid (12.53 ± 1.00 mg/g DW), neophytadiene (8.42 ± 0.51 mg/g DW), and α-linolenic acid (1.27 ± 0.005 mg/g DW) were among the prominent compounds identified. It is plausible to suggest that the abundance of these lipophilic compounds contributes to the remarkable antioxidative and anti-inflammatory potential exhibited by the FD-CM extract. Our results provide insights into the preferential drying methods and extraction solvents for producing M. nummuloides-based products with enhanced antioxidant and anti-inflammatory activity.

Essential oil from the stems of Croton kongensis Gagnep.: chemical composition, antimicrobial and anti-inflammatory activities

The current study describes the chemical compositions, antimicrobial and anti-inflammatory activities of the essential oil (EO) from the stems of Croton kongensis Gagnep., collected from two wild-growing populations in Thanh Hoa province, Vietnam, namely Nhu Xuan and Thuong Xuan. According to gas chromatography/mass spectrometry (GC/MS) analyses, the major components in the Nhu Xuan EO sample were (E)-caryophyllene (10.62%), bornyl acetate (9.81%), (E)-nerolidol (9.15%), and linalool (8.10%). In contrast, linalool (15.05%), bornyl acetate (9.52%), (E)-caryophyllene (7.91%), bicyclogermacrene (7.36%), 1,8-cineole (6.53%), and (E)-nerolidol (6.00%) were the dominant components in the Thuong Xuan EO sample. The two EO samples were screened for antimicrobial activity against a panel of pathogenic bacteria and fungi using a broth microdilution assay and showed promising antimicrobial activities with minimum inhibitory concentration (MIC) ≤ 200 μg/mL. Among them, the Nhu Xuan EO sample exhibited better activity against four bacteria and three fungi compared to the Thuong Xuan EO sample, which only showed activity against one bacterium and three fungi. Additionally, the anti-inflammatory study demonstrated that the EO samples from Nhu Xuan and Thuong Xuan inhibited nitric oxide production in lipopolysaccharide-stimulated RAW 264.7 macrophage cells, with IC50 values of 105.71 ± 0.96 μg/mL and 94.93 ± 1.31 μg/mL, respectively. In conclusion, this is the first report describing the antimicrobial and anti-inflammatory effects of the EO from C. kongensis stems. Our results suggest that the EO from C. kongensis stems in both regions could be potentially utilized as new natural antimicrobial and anti-inflammatory ingredients in the medicine and food industries.

Bioactivity-Guided High Performance Counter-Current Chromatography and Following Semi-Preparative Liquid Chromatography Method for Rapid Isolation of Anti-Inflammatory Lignins from Dai Medicinal Plant, Zanthoxylum acanthopodium var. timbor.

The development of Dai medicine is relatively slow, and Zanthoxylum has great economic and medicinal value. It is still difficult to obtain medicinal components from the low-polarity parts of Zanthoxylum belonging to Dai medicine. In this study, we introduced one simple and quick strategy of separating target compounds from the barks of Z. acanthopodium var. timbor by high-performance countercurrent chromatography (HPCCC) with an off-line anti-inflammatory activity screening mode. The development of this strategy was based on the TLC-based generally useful estimation of solvent systems (GUESS) method and HPCCC in combination. This paper presented a rapid method for obtaining target anti-inflammatory compounds. Three lignins were enriched by HPCCC with an off-line inhibition mode of nitric oxide production in lipopolysaccharide-stimulated RAW264.7 macrophage cells, using petroleum ether-ethyl acetate-methanol-water (3:2:3:2) as the solvent system. The results showed that this method was simple and practical and could be applied to trace the anti-inflammatory components of the low-polarity part in Dai medicine.

Dimeric 2-(2-phenethyl)chromones from agarwood of Aquilaria filaria

Four new 2-(2-phenethyl)chromone dimers (1–4) were isolated from EtOAc extract of agarwood originating from Aquilaria filaria from Philippines. Their structures were elucidated by spectroscopic analysis (1D and 2D NMR, and HRESIMS) and comparison of the experimental and computed ECD curves. Compounds 1–4 exhibited inhibition of nitric oxide production in lipopolysaccharide-stimulated RAW264.7 cells with IC50 values in the range from 33.94 to 57.53 μM.

Anti-inflammatory flavonoid derivatives from the heartwood of Dalbergia odorifera T. Chen

Two pairs of flavonoid enantiomers (1a/1b and 2a/2b) together with three known analogues (3–5) were isolated from the heartwood of Dalbergia odorifera T. Chen. Their structures were elucidated by extensive spectroscopic analysis (1 D and 2 D NMR, UV, IR, and HRMS) and experimental and calculated ECD data. Compound 2 features an unusual 2-methyl-3(2H)-furanone moiety forming the C-ring of flavonoid, and its putative biosynthetic pathway is also proposed. Compounds 3‒5 exhibited significant inhibition of nitric oxide production in lipopolysaccharide-stimulated RAW264.7 cells with IC50 values of 14.7 ± 0.3 μM, 40.2 ± 1.1 μM, and 3.2 ± 0.1 μM, respectively.

Comparative Evaluation of Antioxidant and Anti-Inflammatory Activity of Active Compounds Identified in Ardisia elliptica Extracts from Different Plant Parts.

Ardisia elliptica Thunb. (AE) has been used as food and in traditional medicine to prevent and treat fever, diarrhea, chest pain, liver poisoning, and parturition complications in Southeast Asian countries. This study focused on phytochemical constituents of AE extracts and their antioxidant and anti-inflammatory activity in vitro by evaluating nitric oxide production, and DPPH and FRAP radical scavenging activity. The bioactive compounds from different plant parts, including old leaves, young leaves, flowers, roots, and fruits, were identified. The results showed the highest phenolic and flavonoid content in the root extract among all extracts, which resulted in the most potent free radical scavenging activity revealed by the DPPH and FRAP assay. The roots and flowers showed the highest bergenin (3.36±0.22 mg/g dry weight) and quercetin (2.99±0.10 mg/g dry weight) content, respectively. In contrast, embelin was found only in the fruits. Interestingly, AE extracts significantly suppressed the mRNA expression of inducible nitric oxide synthase, leading to inhibition of nitric oxide production in lipopolysaccharide-stimulated RAW 264.7 macrophages. Conclusively, the results suggest the natural products of AE extracts as effective antioxidant and anti-inflammatory agents that can be utilized for food and pharmaceutical applications.

Filarones A and B, new anti-inflammatory dimeric 2-(2-phenethyl)chromones from agarwood of Aquilaria filaria

• Two previously unreported dimeric 2-(2-phenethyl)chromones were isolated from agarwood of Aquilaria filaria. • Compounds 1 and 2 were two flindersia 2-(2-phenylethyl) chromone dimers linked by an ether bond. • Compounds 1 and 2 exhibited significant inhibition of nitric oxide production in LPS-stimulated RAW264.7 cells. Two new dimeric 2-(2-phenethyl)chromones (filarones A and B) were isolated from agarwood of Aquilaria filaria (Thymelaeaceae) by LC–MS-guided fractionation procedure. Their structures were established based on spectroscopic methods including HRESIMS, one and two dimensional NMR. Filarones A and B exhibited significant inhibition of nitric oxide production in lipopolysaccharide-stimulated RAW264.7 cells with IC 50 values of 11.33 ± 1.76 μ M and 10.69 ± 0.18 μ M. Quercetin and indomethacin were used as positive controls with IC 50 values of 8.22 ± 0.80 μ M and 35.40 ± 1.77 μ M, respectively.

Bioactive Polyketides From the Potato Endophytic Fungus Aspergillus carneus

A previously undescribed polyketide (1) and 3 known analogs (2-4) were obtained from cultures of the potato endophytic fungus Aspergillus carneus. The structures were elucidated on the basis of extensive nuclear magnetic resonance spectroscopic data. The absolute configuration of 1 was further determined by electronic circular dichroism and optical rotation calculations. Compounds 1-4 showed moderate antifungal activity against plant pathogens. Compounds 1, 2, and 4 inhibited nitric oxide production in lipopolysaccharide-stimulated RAW264.7 cells, with half-maximal inhibitory concentration values of 13.36, 30.16, and 51.47 µM, respectively. Compound 4 showed effective antioxidant activity.

Anti-inflammatory Metabolites from Chaetomium nigricolor.

Twelve metabolites were obtained from the culture media of Chaetomium nigricolor, including a new furan derivative, methyl succinyl Sumiki's acid (1), and two new atropisomers of the previously reported bis-naphtho-γ-pyrones, (aS)-asperpyrone A and (aS)-fonsecinone A (2 and 3). The structures were elucidated by spectroscopic, chemical, and chiroptical techniques. Compounds 2 and 3 inhibited nitric oxide production in lipopolysaccharide-stimulated RAW 264.7 macrophages. Compound 2 was found to inhibit nuclear factor-kappa B and c-Jun N-terminal kinase activation, in turn suppressing pro-inflammatory mediators and cytokines including nitric oxide, prostaglandin E2, interleukin (IL)-1β, tumor necrosis factor-α, IL-6, and IL-12.

Macrocyclic Diterpenoids from Euphorbia helioscopia and Their Potential Anti-inflammatory Activity.

Guided by 1H NMR spectroscopic experiments using the aromatic protons as probes, 11 macrocyclic diterpenes (1-11) were isolated from the aerial parts of Euphorbia helioscopia. Their full three-dimensional structures, including absolute configurations, were established unambiguously by spectroscopic analysis and single-crystal X-ray crystallographic experiments. Among the isolated compounds, compound 1 is the third member thus far of a rare class of Euphorbia diterpenes featuring an unusual 5/10 fused ring system, and 2-4 are new jatrophane diterpenes. Based on the NMR data of the jatrophane diterpenes obtained in this study as well as those with crystallographic structures reported in the literature, the correlations of the chemical shifts of the relevant carbons and the configurations of C-2, C-13, and C-14 of their flexible macrocyclic ring were considered. Moreover, the anti-inflammatory activities of 1-11 were investigated by monitoring their inhibitory effects on nitric oxide production in lipopolysaccharide-stimulated RAW 264.7 cells. Compound 1 showed an IC50 of 7.4 ± 0.6 μM, which might be related to the regulation of the NF-κB signaling pathway by suppressing the translocation of the p65 subunit and the consequent reduction of IL-6 and TNF-α secretions.

Lignans and Terpenoids From the Fruits of Vitex kwangsiensia and Their Inhibitory Activity on Nitric Oxide Production in Macrophages

Phytochemical investigation of the fruits from Vitex kwangsiensia led to the isolation of 18 constituents, including 6 lignans (1-6), 3 diterpenoids (7-9), 2 sesquiterpenoids (10, 11), 2 triterpenoids (16, 17), and 5 other compounds. Their structures were identified by spectroscopic methods (mass spectroscopy, infrared, UV, 1-dimensional and 2-dimensional nuclear magnetic resonance) and comparison with reported data in the literatures. Among these constituents, compound 1 was a new lignan and named as vitekwangin A. The spectroscopic data of 2 was reported herein for the first time, and given a trivial name vitekwangin B. The isolated lignans, diterpenoids, and sesquiterpenoids were evaluated for their inhibitory activity to prevent nitric oxide production in lipopolysaccharide-stimulated RAW264.7 macrophages.

Punicagranine, a new pyrrolizine alkaloid with anti-inflammatory activity from the peels of Punica granatum

A novel pyrrolizine alkaloid, 5-(furan-3-carbonyl)-2,3-dihydro-1H-pyrrolizine-7-carboxylic acid, was isolated from the peels of Punica granatum. Its structure was elucidated on the basis of 1D- and 2D-NMR and HR-ESI-MS spectra and then confirmed by X-ray diffraction. It represents a new type of natural pyrrolizine alkaloid, named, punicagranine (1). 1 showed inhibition of nitric oxide production in lipopolysaccharide-stimulated RAW 264.7 cells. To our knowledge, the pyrrolizine alkaloid skeleton was reported in natural products for the first time.

LC-MS-guided isolation of anti-inflammatory 2-(2-phenylethyl)chromone dimers from Chinese agarwood (Aquilaria sinensis)

Fifteen previously undescribed 2-(2-phenylethyl)chromone dimers, along with two known analogues were isolated from Chinese agarwood (Aquilaria sinensis) by a LC-MS-guided fractionation procedure. Their structures were elucidated on the basis of spectroscopic and spectrometric data (1D and 2D NMR, IR, and HRESIMS). The isolated compounds exhibited significant inhibition of nitric oxide production in lipopolysaccharide-stimulated RAW264.7 cells with IC50 values in the range 0.6–37.1 μM.

Chemical Constituents of Apios americana Tubers and Their Inhibitory Activities on Nitric Oxide Production in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages.

Apios americana is an important food crop producing edible tubers with high nutritional and medicinal values and is widely cultivated in many countries. Despite its usefulness, research on its secondary metabolites and biological activities has been limited. In the present study, a new coumaronochromone, (2 R,3 S)-3,7,4'-trihydroxy-5-methoxycoumaronochromone (1), and two new isoflavone glucosides, 7,2',4'-trihydroxy-5-methoxyisoflavone-4'- O-β-d-glucopyranoside (3) and 5,7,4'-trihydroxyisoflavone-7- O-β-d-gentiotrioside (5), were isolated from the tubers of A. americana via chromatographic separation. Seventeen known compounds (2, 4, and 6-20) were also obtained from this plant part. The chemical structures of 1, 3, and 5 were determined by the interpretation of spectroscopic data. The absolute structure of the new compound 1 was established from experimental and calculated electronic circular dichroism spectra. This is the first study to determine the absolute configuration of a 3-hydroxycoumaronochromone derivative. The potential anti-inflammatory activity of the 20 isolates obtained was evaluated by measuring their inhibitory effects on nitric oxide production in lipopolysaccharide-stimulated RAW 264.7 macrophages. Among the isolates, seven compounds (1, 3, 6-8, 15, and 20) showed substantial inhibition of nitric oxide production in RAW 264.7 cells, with the most active being compound 1 (IC50 value of 0.38 ± 0.04 μM).

Multiflorumisides A-G, Dimeric Stilbene Glucosides with Rare Coupling Patterns from the Roots of Polygonum multiflorum.

Multiflorumisides A-G (1-7), seven new dimeric stilbene glucosides with two rare coupling patterns, were isolated from the roots of Polygonum multiflorum. The structures of these new dimeric stilbene glucosides were elucidated through comprehensive spectroscopic and chemical analyses. The absolute configurations of 3 and 5-7 were established by comparing their experimental and quantum-chemical ECD data. Putative biosynthetic pathways toward the dimers and their suppressive effects against nitric oxide production in lipopolysaccharide-stimulated RAW264.7 cells are also discussed.

Anti-inflammatory Dimeric 2-(2-Phenylethyl)chromones from the Resinous Wood of Aquilaria sinensis.

Sixteen new 2-(2-phenylethyl)chromone dimers, including four pairs of enantiomers (1a/1b, 3a/3b, 6a/6b, and 8a/8b), along with eight optically pure analogues (2, 4, 5, 7, and 9-12) were isolated from the resinous wood of Aquilaria sinensis. Their structures were determined by extensive spectroscopic analysis (1D and 2D NMR, UV, IR, and HRMS) and experimental and computed ECD data. Compounds 1-10 feature an unusual 3,4-dihydro-2 H-pyran ring linkage connecting two 2-(2-phenylethyl)chromone monomeric units, while compounds 11 and 12 possess an unprecedented 6,7-dihydro-5 H-1,4-dioxepine moiety in their structures. A putative biosynthetic pathway of the representative structures via a diepoxy derivative of a chromone with a nonoxygenated A-ring is also proposed. Compounds 1a/1b, 2, 3a/3b, 5, 7, 8a/8b, and 10-12 exhibited significant inhibition of nitric oxide production in lipopolysaccharide-stimulated RAW264.7 cells with IC50 values in the range 7.0-12.0 μM.