NO Production Inhibitory Activity Research Articles

Four Unidentified Compounds Isolated from the Stem Barks of Aphanamixis polystachya and Their NO Production Inhibition in LPS Activated RAW 264.7 Cells.

Phytochemical study on the methanol extract of the stem barks of Aphanamixis polystachya led to the isolation of four previously undescribed ( 1-4) and ten known compounds (5-14). Their chemical structures were elucidated to be 11-methoxysawaranospiroride C (1), 6α,9S,10,13-tetrahydroxymegastigmane-3-one (2), 11-hydroxyaphanamixin B (3), (2Z,6E,13E)-2,6,13-triene-11,15-dihydroxyphytanic acid (4), cinnacasside D (5), cinnacasside E (6), vilsonol F (7), (3S,5R,6S,7E,9R)-3,5,6,9-tetrahydroxy-7-en-megastigmane (8), (3S,5R,6R,7E,9R)-3,6,9,10-tetrahydroxy-7-en-megastigmane (9), citroside A (10), threo-1-(3,4,5-trimethoxyphenyl)-1,2,3-propanetriol (11), 3,4,5-trimethoxyphenyl-1-O-β-D-glucopyranoside (12), p-coumaric acid (13), ferulic acid (14) by HR-ESI-MS, ECD, 1D-, and 2D-NMR spectra. Compounds 1, 3, 4, and 9 showed NO production inhibitory activity in LPS activated RAW 264.7 cells with IC50 values of 42.0, 67.9, 20.5, and 78.6 µM, respectively, while the remaining compounds were inactive with IC50 values over 100 µM.

Tributelosides A-D: Four Undescribed Spirostan Glycosides Isolated from the Branches and Leaves of Tribulus terrestris with Their NO Production Inhibitory Activity in LPS Activated RAW 264.7 Cells.

Four undescribed spirostan glycosides, (25S)-5α-spirostan-12-one-2α,3β-diol-3-O-β-D-glucopyranosyl-(1→4)-β-D-galactopyranoside (1), (25S)-5α-spirostan-12-one-2α,3β-diol-3-O-β-D-galatopyranosyl-(1→2)-β-D-glucopyranosyl-(1→4)-β-D-galactopyranoside (2), (25S)-5α-spirostan-12-one-2α,3β-diol-3-O-β-D-glucopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→3)]-β-D-glucopyranosyl-(1→4)-β-D-galactopyranoside (3), and hecogenin 3-O-β-D-glucopyranosyl-(1→3)-[β-D-xylopyranosyl-(1→2)]-β-D-glucopyranosyl-(1→4)-[α-L-rhamnopyranosyl-(1→2)]-β-D-galactopyranoside (4), together with eleven known compounds (5-15) were isolated from the branches and leaves of Tribulus terrestris. Their chemical structures were established through spectroscopic methods. including HR-ESI-MS, 1D-, and 2D-NMR spectra. Preliminary biological evaluation on NO production inhibitory activity in LPS activated RAW 264.7 cells showed that compounds 1-3, 5, and 6 had significant inhibitory effects with IC50 values ranging from 2.4 to 18.3 µM, compared to that of the positive control compound, dexamethazone (IC50 13.6 µM).

Undescribed Lignanamide and Flavone C-Glucoside Isolated from the Aerial Parts of Piper Samentosum with NO Production Inhibitory Activity in LPS Activated RAW 264.7 Cells.

In this study, two undescribed compounds (1 and 2), together with eight known compounds (3-10) were isolated from the aerial parts of Piper samentosum by various chromatography methods. Their chemical structures were determined to be 7'''-oxolyciumamide N (1), vitexin 2''-O-β-D-(6'''-feruloyl)-glucopyranoside (2), 1,2-dihydro-6,8-dimethoxy-7-hydroxy-1-(3,4-dihydroxyphenyl)-N1,N2-bis-[2-(-hydroxyphenyl)ethyl]-2,3-napthalene dicarboamide (3), vitexin 6''-O-β-D-glucopyranoside (4), vitexin 2''-O-α-L-rhamnopyranoside (5), methyl 2-hydroxybenzoate-2-O-β-D-apiofuranosyl-(1→2)-O-β-D-glucopyranoside (6), ficuside G (7), methyl 2-O-β-D-glucopyranosylbenzoate (8), methyl 2,5-dihydroxybenzoate-5-O-β-D-glucopyranoside (9), and 3,7-dimethyloct-1-ene-3,6,7-triol 6-O-β-D-glucopyranoside (10) by spectroscopic data analysis including HR-ESI-MS, 1D-, and 2D-NMR spectra. Compounds 1-5 inhibited nitric oxide production in LPS-stimulated RAW264.7 macrophages with the IC50 values of 27.62, 74.03, 38.54, 70.39, and 44.95 μM, respectively. The NMR data of 9 were firstly reported herein.

Acemannan과 aloesin의 혼합 비율에 따른 대식세포에서의 항염증 효과

This study aims to confirm the anti-inflammatory activities of acemannan and aloesin, which have been studied for various efficacies at various mixed sample ratios. The mixed samples were mixed at a ratio of 1:1 (AA-1), 1:2 (AA-2), 1:3 (AA-3), 2:1 (AA-4), and 3:1 (AA-5). Seven samples were evaluated for their cytotoxic ability on macrophages, and the results showed that all cell viability was over 90% at a concentration of 100 μg/mL. First, due to the NO production inhibitory activity, a better inhibitory effect was achieved when using a mixed sample rather than a single material. Afterward, the activity of inhibiting the production of PGE2, TNF-α, and IL-6 was confirmed using a mixed sample. It was confirmed that AA-2 had the best inhibitory activity on producing PGE2, TNF-α, and IL-6 rather than AA-1, AA-3, AA-4, and AA-5. For this reason, experiments were conducted using AA-2 to determine the protein expression levels of iNOS and COX-2, which are inflammation-related proteins. It was confirmed that AA-2 inhibited iNOS and COX-2 protein expression by 25.01% and 27.27%, respectively, compared to the LPS-alone treatment group. In conclusion, the mixed sample of acemannan and aloesin is judged to have anti-inflammatory activity and can potentially to be used as a functional material.

Phytoconstituents of Androstachys johnsonii Prain Prevent Reactive Oxygen Species Production and Regulate the Expression of Inflammatory Mediators in LPS-Stimulated RAW 264.7 Macrophages.

According to a survey, the medicinal use of Androstachys johnsonii Prain is kept secret by traditional healers. Considering that inflammation and oxidative stress are major risk factors for the progression of various chronic diseases and disorders, we resolved to investigate the antioxidant and anti-inflammatory potentials of A. johnsonii using in vitro and cell-based assays. The antioxidant activity of A. johnsonii hydroethanolic leaf extract (AJHLE) was evaluated using the ABTS, DPPH, and FRAP assays. Its cytotoxic effect was assessed on RAW 264.7 macrophages using an MTT assay. Then, its anti-inflammatory effect was evaluated by measuring the NO production and 15-LOX inhibitory activities. Moreover, its preventive effect on ROS production and its regulatory effect on the expression of pro-inflammatory mediators such as IL-1β, IL-10, TNF-α, and COX-2 were determined using established methods. AJHLE strongly inhibited radicals such as ABTS•+, DPPH•, and Fe3+-TPTZ with IC50 values of 9.07 µg/mL, 8.53 µg/mL, and 79.09 µg/mL, respectively. Additionally, AJHLE induced a significant (p < 0.05) cytotoxic effect at 100 µg/mL, and when tested at non-cytotoxic concentrations, it inhibited NO and ROS production in LPS-stimulated RAW 264.7 macrophages in a concentration-dependent manner. Furthermore, AJHLE showed that its anti-inflammatory action occurs via the inhibition of 15-LOX activity, the downregulation of COX-2, TNF-α, and IL-1β expression, and the upregulation of IL-10 expression. Finally, chemical investigation showed that AJHLE contains significant amounts of procyanidin, epicatechin, rutin, and syringic acid which support its antioxidant and anti-inflammatory activities. These findings suggest that A. johnsonii is a potential source of therapeutic agents against oxidative stress and inflammatory-related diseases.

Anti-inflammatory Polyketides from an Endophytic Fungus Chaetomium sp. UJN-EF006 of Vaccinium bracteatum.

Seven new polyketides including three chromone derivatives (1-3) and four linear ones incorporating a tetrahydrofuran ring (4-7), along with three known compounds (8-10), were obtained from the fermentation of an endophytic fungus (Chaetomium sp. UJN-EF006) isolated from the leaves of Vaccinium bracteatum. The structures of these fungal metabolites have been elucidated by spectroscopic means including MS, NMR and electronic circular dichroism. A preliminary anti-inflammatory screening with the lipopolysaccharide (LPS) induced RAW264.7 cell model revealed moderate NO production inhibitory activity for compounds 1 and 4. In addition, the expression of three LPS-induced inflammatory factors IL-6, iNOS and COX-2 was also blocked by 1 and 4.

Ecdysterols and triterpene glycoside from Achyranthes aspera L. and their NO production inhibitory activity

Phytochemical study on the methanolic extract of the aerial parts of Achyranthes aspera led to the isolation of six compounds including five ecdysterols, makisterone A (1), achyranthesterone A (2), 24(28)-dehydromakisterone A (3), podecdysone C (4), 20-hydroxyecdysone (5), and 3-O-α-L-rhamnopyranosyl-(1→3)-β-D-glucuronopyranosyl-oleanolic acid 28-O-β-D-glucopyranosyl ester (6). Compounds 1-5 were first isolated from A. aspera. Their chemical structures were identified by 1D and 1D NMR spectra in comparison with the published data in literature. Al five ecdysterol significantly showed NO production inhibition effects in LPS-activated RAW264.7 cells with the IC50 values ranging from 27.21 to 40.47 µM compared to that of positive control compound, L-NMMA, 32.24 µM.

Undescribed sesquiterpenoids with NO production inhibitory activity from oleo-gum resin of Commiphora myrrha

Six undescribed cadinane sesquiterpenoids (1−6), two undescribed guaiane sesquiterpenoids (7−8), and an undescribed germacrane sesquiterpenoid (9) were isolated from the oleo-gum resin of Commiphora myrrha. Their structures were determined by the analysis of 1D/2D NMR and HRESIMS data, as well as quantum chemical ECD and NMR calculations. All the sesquiterpenoids were evaluated for their NO production inhibitory activity in LPS-stimulated RAW 264.7 mouse monocyte-macrophages. The results revealed that commiphone A (1) and commipholide D (7) exhibited significant inhibitory effect on NO generation with IC50 values of 18.6 ± 2.0 and 37.5 ± 1.5 μM, respectively. Furthermore, 1 and 7 dose-dependently inhibited the mRNA expression of inflammatory cytokines IL-1β, IL-6 and TNF-α induced by LPS in the RAW264.7 cells, indicating that 1 and 7 possess potent anti-inflammatory activity in vitro.

Four Undescribed compounds Isolated from the Aerial Parts of Phyllanthus cochinchinensis with Antimicrobial Activity and NO Production Inhibitory Activity in LPS Activated RAW 264.7 Cells.

Four previously undescribed compounds named phyllancosides A and B (1 and 2), and phyllancochines A and B (3 and 4) together with ten known compounds (5-14) were isolated from the aerial parts of Phyllanthus cochinchinensis Spreng. Their chemical structures were elucidated on the basis of comprehensive analysis of IR, HR-ESI-MS, 1D and 2D NMR spectra. The absolute configurations of 1 and 2 were determined by electronic circular dichroism (ECD) spectra. Compounds 3, 4, and 10 showed antimicrobial activity against E. faecalis, S. aureus, and B. cereus with the MIC values in range of 32-256 μg/mL. Compound 11 inhibited E. faecalis and B. cereus, and 7 inhibited S. aureus with the MIC values in range of 64-128 μg/mL. In addition, compounds 1, 3, 4, 8, and 9 showed significantly NO production inhibitory activity in LPS activated RAW 264.7 cells with IC50 values ranging from 36.57 to 56.34 μM.

Chemical constituents of Syzygium cerasiforme leaves and their nitric oxide inhibitory activity in LPS‐activated RAW264.7 cells

AbstractA phytochemical study of the methanolic extract of Syzygium cerasiforme leaves led to the isolation of seven compounds (1–7). Their chemical structures were identified as benzyl‐6ʹ‐O‐galloyl‐β‐d‐glucopyranoside (1), 3‐methoxy‐4‐hydroxyphenol 1‐O‐β‐D‐(6ʹ‐O‐galloyl)‐glucopyranoside (2), betulonic acid (3), 3‐epi‐betulinic acid (4), 7α‐methoxy‐stigmast‐5‐en‐3β‐ol (5), dihydrodehydrodiconiferyl alcohol (6), and secoisolariciresinol (7), by HR‐ESI‐MS, 1D‐ and 2D NMR spectra, and by comparison with the previously published data. Compounds 1–7 exhibited NO production inhibitory activity in LPS‐activated RAW264.7 cells with IC50 values of 33.17, 25.51, 51.09, 54.17, 39.54, 6.98, and 58.12 µm, respectively, compared to that of the positive control compound, L‐NMMA, which showed an IC50 of 32.5 µm. This is the first report on these compounds from S. cerasiforme.

Bioactive monoterpenoids and acetophenones from the aerial parts of Eupatorium fortunei

Thirty-nine thymol and acetophenone derivatives, including eight pairs of enantiomers, were isolated from the aerial parts of Eupatorium fortunei. Their structures were assigned by detailed analyses of spectroscopic data and NMR calculations based on density functional theory, with 18 ones (1a/1b−14) being previously undescribed compounds. While the absolute configurations of 1a/1b, 2a/2b, 4, 6a/6b, 7, 11a/11b and 15a/15b−18a/18b were established by calculations of electronic circular dichroism data, that of 14 was determined by modified Mosher's method. Compounds 1a/1b and 2a/2b represent a previously unreported type of monoterpenoid dimers via an amide linkage, and compound 3 is a monoterpene-phenylpropanoid hybrid connected through an ester bond. Among the known molecules, the formerly mis-assigned structures of 15a/15b and 22 were revised, and pure natural enantiomers of 16a/16b−18a/18b were reported for the first time. Selective compounds showed antiradical and NO production inhibitory activities in the preliminary biological screening. Compound 31 was further demonstrated to alleviate oxidative stress by activating Nrf2 signaling pathway.

롱빈 콩 에탄올추출물의 항산화 및 NO 생성 억제 효과

In order to prepare basic data for examining the development of longbean (Vigna unguiculata ssp. sesquipedalis) as a possible natural material for functional cosmetics, the study investigated the yield, total polyphenol content, antioxidant, and the NO production inhibitory effects of an ethanol-produced extract of longbean soybean parts. As a result, the yield of the longbean ethanol extract was found to be 14.93% and the total polyphenol content was confirmed to be 4.36±0.473 mg/g. When the antioxidant effect was measured by the electron donating ability and the ABTS radical scavenging activity, the electron donating ability tended to increase as the concentration increased, and showed a high scavenging ability of 92.32% at a concentration of 1,000 μg/mL. And the ABTS radical scavenging activity showed a scavenging effect of 99.81% at a concentration of 100 μg/ml, and the scavenging effect was equivalent to that of vitamin C. As a result of measuring the antioxidant activity, the ABTS radical scavenging ability showed a higher scavenging effect than the electron donating ability. The cell viability of RAW 264.7 cells was 99.82-100.28% up to the concentration of 500 μg/ml, and cytotoxicity was not observed up to 500 μg/ml. The NO production inhibitory activity showed a significant decrease to 81.84% at a concentration of 500 μg/ml. Therefore, it is considered that the long bean ethanol extract can be used as a natural material for cosmetics. In future studies, it is necessary to demonstrate the physiological activity, such as anti-inflammatory mechanisms, whitening, and wrinkle improvement through component analysis.

C21 steroidal glycosides from the root bark of Periploca sepium and their NO production inhibitory and cytotoxic activity

A series of C21 steroidal glycosides were isolated from the root bark of Periploca sepium, including a new compound, perisepiumoside A1 (1), and six known compounds (2-7). Their structures were elucidated by analysis of HR-ESI-MS, and 1D and 2D NMR spectroscopic data. All these compounds were tested for their NO production inhibitory activity in LPS-stimulated RAW 264.7 cells. Results showed that these C21 steroidal glycosides could remarkably inhibit NO production, particularly 1 and 2 with IC50 values of 30.81 ± 0.18 μM and 44.39 ± 0.21 μM, respectively. In addition, the cytotoxicity of these compounds was measured on A549, MCF-7, and HeLa cancer cell lines. Among them, compounds 1 and 7 displayed cytotoxicity against the A549 cell line with IC50 values of 28.41 ± 0.12 μM and 39.06 ± 0.05 μM, respectively.

Phenolic Constituents of Elsholtzia blanda Benth

Objectives: Elsholtzia blanda Benth. is a Vietnamese medicinal plant, used to treat inflammation, cough, and flu. Therefore, this current study focused on the isolation of the active ingredients from this plant and their NO production inhibitory activity. Methods: E. blanda aerial parts were dried, minced, and ultrasonically extracted with methanol and further separated by various chromatographic methods. The chemical structures of isolates were revealed by nuclear magnetic resonance, high resolution lectrospray ionization mass spectrometry, and circular dichroism spectra. The nitric oxide inhibition effects were evaluated in lipopolysaccharide-activated RAW264.7 cells. Results: Ten phenolic components [1 new (1) and 9 known (2-10)] were isolated and their chemical structures determined. Compound 5 (innovanoside, IC50 = 19.23 ± 1.43 µM) and 6 (maltol- O- β-D-glucopyranoside, IC50 = 21.73 ± 1.27 µM) showed the highest activity, and 2-4 showed moderate inhibition (IC50 = 47.36 ∼ 57.29 µM). The other compounds were inactive with IC50 values &gt;100 µM. Conclusions: One previously undescribed and 9 known phenolic components isolated from E. blanda have been screened for their nitric oxide inhibitory activity. Compounds 2-6 showed moderate to significant inhibition with IC50 values of 19.23∼57.29 µM.

Four New Flavonoid C-Glycosides Isolated from Achyranthes aspera and Their Nitric Oxide Production Inhibitory Activities.

A chemical study of the methanol extract of the aerial parts of Achyranthes aspera led to the isolation of four new flavonoid C-glycosides (1-4) along with eight known analogs (5-12). Their structures were elucidated by a combination of spectroscopic data analysis, HR-ESI-MS, 1D and 2D NMR spectra. All the isolates were evaluated their NO production inhibitory activity in LPS-activated RAW264.7 cells. Compounds 2, 4, and 8-11 showed significant inhibition with IC50 values ranging from 25.06 to 45.25 μM, compared to that of the positive control compound, L-NMMA, IC50 value of 32.24 μM, whereas the remaining compounds were weak inhibitory activity with IC50 values over 100 μM. This is the first report of 7 from Amaranthaceae family, and 11 from the genus Achyranthes.

The chemical composition from the fruits of Pandanus tonkinensis, their NO production inhibitory and lipid peroxidation inhibitory activities

AbstractSeven known compounds, including two neolignans, four lignans, and one flavane were isolated from the methanol extract of Pandanus tonkinenis fruits. Their structures were determined to be ficusal (1), vladinol F (2), medioresinol (3), syringaresinol (4), lariciresinol (5), secoisolariciresinol (6), and luteoliflavan (7) by analysis of MS and NMR data as well as by comparison of their spectral data with those reported in the literature. All compounds were evaluated for NO production inhibitory and lipid peroxidative inhibitory activities. Compounds 5 and 7 showed significant NO production inhibitory with IC50 values of 5.3±0.4 and 7.1±0.4 μM, respectively. Compounds 6 and 7 showed significant lipid peroxidative inhibitory activity with IC50 values of 20.2±1.7 and 26.2±3.6 μM, respectively.

Cytochalasans from the Endophytic Fungus Aspergillus sp. LE2: Their Structures, Antibacterial and NO Production Inhibitory Activities

Six new cytochalasans-namely, aspergicytochalasins A-F (1-6)-together with five known analogs were isolated and characterized from the endophytic fungus Aspergillus sp. from the medicinal plant Lonicera japonica. The structures of the new compounds were established by NMR and MS methods as well as single crystal X-ray diffractions. Compounds 3 and 4 showed weak antibacterial activities to Staphylococcus aureus, with MIC values of 128 and 64 μg/mL, respectively. Compounds 1, 3, 5 and 6 showed inhibitory activities on NO production, with IC50 values less than 40 μM.

Chemical constituents from the arial of Crinum asiaticum L

Crinum asiaticum L. (locally known Nang hoa trang), belonging to the Crinum genus of the Amaryllidaceae family, has been widely used in oriental and traditional medicine around the world for the treatment of various diseases and exerts diverse promising pharmacological effects. To find out new inhibitors for the treatment of inflammatory and Alzheimer’s diseases, a methanol extract of the aerial part of C. asiaticum was investigated to afford six compounds, chrysophanol (1), emodin (2), physcion (3), thymine (4), thymidine (5), and 5-(hydroxymethyl)-2-furfural (6) by using various chromatographic separations. Their structures were identified by the extensive analysis of 1H- and 13C-NMR data as well as a comparison with those of previously reported data. This is the first report of these compounds (1-6) from C. asiaticum. In addition, all compounds evaluated the inhibitory activities of NO production and acetylcholinesterase (AChE). Among them, emodin (2) significantly inhibited NO production with an IC50 of 5.13±0.42 μM. This result indicated that emodin (2) may be beneficial in the treatment of anti-inflammatory diseases. Besides, all compounds (1-6) also showed weak inhibitory effects on the AChE enzyme with a range of IC50 values from 82.98±9.19 to 237.63±13.8 µM.

Chemical constituents from the arial of Crinum asiaticum L.

Crinum asiaticum L. (locally known Nang hoa trang), belonging to the Crinum genus of the Amaryllidaceae family, has been widely used in oriental and traditional medicine around the world for the treatment of various diseases and exerts diverse promising pharmacological effects. To find out new inhibitors for the treatment of inflammatory and Alzheimer’s diseases, a methanol extract of the aerial part of C. asiaticum was investigated to afford six compounds, chrysophanol (1), emodin (2), physcion (3), thymine (4), thymidine (5), and 5-(hydroxymethyl)-2-furfural (6) by using various chromatographic separations. Their structures were identified by the extensive analysis of 1H- and 13C-NMR data as well as a comparison with those of previously reported data. This is the first report of these compounds (1-6) from C. asiaticum. In addition, all compounds evaluated the inhibitory activities of NO production and acetylcholinesterase (AChE). Among them, emodin (2) significantly inhibited NO production with an IC50 of 5.13±0.42 μM. This result indicated that emodin (2) may be beneficial in the treatment of anti-inflammatory diseases. Besides, all compounds (1-6) also showed weak inhibitory effects on the AChE enzyme with a range of IC50 values from 82.98±9.19 to 237.63±13.80 µM.

Sesquiterpene lactones from Artemisia vulgaris L. as potential NO inhibitors in LPS-induced RAW264.7 macrophage cells.

Twelve new guaianolide sesquiterpene lactones (1–12), along with ten known analogs (13–22) were isolated from an EtOH extract of the dried aerial parts of Artemisia vulgaris L. The new structures were elucidated via abundant spectroscopic data analyses (HRESIMS, IR, 1D, and 2D NMR), and the absolute configurations of these compounds were determined by X-ray crystallography and ECD calculations. The compounds (1−22) were identified as guaiane-type sesquiterpenes with characteristic α-methylene-γ-lactone and α,β-unsaturated carbonyl moieties. All compounds were tested for their inhibitory activity against NO production in lipopolysaccharide-stimulated RAW264.7 macrophages. The isolated sesquiterpenoids dose-dependently exhibited an NO production inhibitory activity by inhibiting the expression of inducible NO oxidase (iNOS) and cyclooxygenase-2 (COX-2) with IC50 values ranging from 1.0 to 3.6 μM. The inhibitory effect on the NO production of the compounds (1–4 and 6–22) is better than that of the positive control (dexamethasone). The different substitutions of compounds on C-8 influence anti-inflammatory effects, as evidenced by the in silico analysis of related binding interactions of new compounds (1–12) with iNOS.