Inhibition Of Nitric Oxide Production Research Articles

Fatty acids in Plantago asiatica seeds are responsible for the production of the pro-inflammatory mediator nitric oxide

Background: Plantago asiatica L. (Plantaginaceae) commonly grows in East Asia, with its seeds (Shazenshi), having been used as diuretic and anti-inflammatory drugs in traditional Japanese medicine. It is not known which constituents of P. asiatica seeds elicit the anti-inflammatory effects, such as reduced expression of the inducible nitric oxide synthase (iNOS) in interleukin (IL)-1β-treated hepatocytes which leads to a reduction in the pro-inflammatory mediator nitric oxide (NO). Objective: To identify the anti-inflammatory constituents of P. asiatica seeds, the anti-inflammatory activity of purified constituents was determined by assaying NO production in IL-1β-treated hepatocytes. Methods: P. asiatica seeds were extracted with 50% methanol and successively fractionated into three crude fractions with ethyl acetate (EtOAc) and n-butanol. The compounds were methylated and analyzed by gas chromatography–mass spectrometry (GC–MS). Primary cultured rat hepatocytes were prepared by collagenase perfusion, and P. asiatica seed extract (PASE), a fraction, or a compound was added to the culture medium with IL-1β and incubated at 37 °C. Potency of each fraction was determined by the Griess method for measuring the levels of nitrite in the medium. Results: PASE suppressed IL-1β-induced NO production without showing cytotoxicity, and an EtOAc-soluble fraction of PASE significantly inhibited NO production. GC–MS analysis detected 26 distinct fatty acids as their methyl esters in this fraction. Among them, three unsaturated fatty acids (linoleic, oleic, and α-linolenic acids) and palmitic acid were abundant. These unsaturated fatty acids are known to reduce NO levels. In contrast, acteoside and aucubin, which are thought to be present in the n-butanol-soluble fraction, showed only a low level of NO production suppression. Conclusion: The EtOAc-soluble fraction of PASE included many fatty acids, which may suppress the production of NO. The results imply that the anti-inflammatory activity of P. asiatica seeds may be produced by three unsaturated fatty acids. Because the fatty acids are abundant in the seeds of medicinal plants, they are likely to contribute to anti-inflammatory activity of the seeds. Keywords: Plantain seed, polyunsaturated fatty acid, Kampo medicine, hepatocyte, nitric oxide.

Development of novel nitric oxide production inhibitors based on the 7H-pyrrolo[2,3-d]pyrimidine scaffold.

Nitric oxide (NO), the smallest signaling molecule known, can be excessively produced by overexpressed inducible nitric oxide synthase (iNOS), and eventually leads to multiple inflammatory related diseases. Thus, reducing the overexpression of NO represents as very potential anti-inflammatory strategy. In current study, a series of compounds were designed and synthesized based on the hybridization of 7H-pyrrolo[2,3-d]pyrimidine and cinnamamide fragments in order to develop novel NO production inhibitors. Among them, compound S2h displayed a vigorous inhibitory activity on NO production with an IC50 value of 3.21 ± 0.67 µM, which was much lower than that of the positive control Nω-nitro-L-arginine (L-NNA, IC50 = 28.36 ± 3.13 µM). Due to its obeying Lipinski's and Veber's rules that guarantee compounds with good oral bioavailability, S2h effectively suppressed the paw swelling in carrageenan-induced mice. Additionally, compound S2h formed clear interactions with iNOS protein according to the docking analysis. Therefore, compounds S2h is a promising lead compound for further development of potent iNOS inhibitors or anti-inflammatory agents.

In-vitro comparative study of solvent effect on antioxidant properties of leaf extracts of Anthocleista djalonensis

This study demonstrated that the extraction solvents play an important role in the extraction of important bioactive compounds with antioxidant properties from the leaves of Anthocleista djalonensis. Anthocleista djalonensis, a plant native to tropical Africa, is believed to have antioxidant properties due to its bioactive composition. The powdered leaves were macerated using local gin, hot distilled water, and analytical ethanol to extract bioactive compounds. The in vitro antioxidant activity of these extracts was assessed using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) scavenging assays, nitric oxide scavenging assay, and hydrogen peroxide scavenging assay. Ascorbic acid was used as a reference. The DPPH and hydrogen peroxide scavenging tests showed concentration-dependent responses. Distilled water had the lowest IC50 value of 0.079 mg/ml and showed the most antioxidant potential in DPPH. Local gin demonstrated significant hydrogen peroxide scavenging ability with an IC50 value of 0.12 mg/ml, surpassing the standard drug. The nitric oxide scavenging assay showed a concentration-dependent increase in both ethanol and local gin extracts. Ethanol and local gin extracts showed maximum antioxidant activity, with ethanol extract showing 22.84% inhibition and local gin extract showing a higher activity of 46.29% inhibition. However, ascorbic acid showed the highest inhibition of nitric oxide production at 57.10%. The study highlights the importance of solvent selection in herbal preparation for optimal health benefits regarding antioxidant activity.

Chemical Profile and Bioactivity of Rubus idaeus L. Fruits Grown in Conventional and Aeroponic Systems.

Raspberry (Rubus idaeus L.) is a fruit of great interest due to its aroma, nutritional properties, and the presence of many bioactive compounds. However, differences among cultivation systems can affect its composition and, consequently, its potential bioactivity. Herein, for the first time, raspberries grown in an aeroponic system were investigated for their chemical profile and antioxidant and anti-inflammatory activity, as well as their enzyme (α-glucosidase and pancreatic lipase) inhibitory properties in comparison to wild and conventionally cultivated fruits. High-performance liquid chromatography coupled with diode array detection (HPLC-DAD) analyses revealed the presence of gallic acid, caffeic acid, chlorogenic acid, p-coumaric acid, ferulic acid, rutin, and catechin in all the samples. The extracts exhibited in vitro anti-inflammatory activity (inhibition of nitric oxide production) regardless of the cultivation method. Of particular interest is the ability of raspberries to inhibit pancreatic lipase. With the exception of the β-carotene bleaching test, the raspberries grown in conventional and aeroponic systems were more active in terms of antioxidants than wild fruits, as evidenced by the ABTS (IC50 in the range 1.6-3.4 μg/mL), DPPH (IC50 in the range 8.9-28.3 μg/mL), and FRAP tests (24.6-44.9 μM Fe(II)/g). The raspberries from aeroponic cultivation were generally able to exert the same bioactivity as those obtained from both conventionally cultivated and wild fruits, supporting the consideration that in the future, this technology could reshape agriculture by mitigating resource constraints, fostering sustainable practices and increasing yields.

Cryptobuchanosides A-G: seven previously undescribed triterpene glycosides from Cryptolepis buchananii R.Br. ex Roem. and Schult. with nitric oxide production inhibition activity.

In this study, nine triterpene glycosides including seven previously undescribed compounds (1-7), were isolated from leaves of Cryptolepis buchananii R.Br. ex Roem. and Schult. using various chromatographic methods. The chemical structures of the compounds were elucidated to be 3-O-β-D-glucopyranosyl-(1 → 6)-β-D-glucopyranosyluncargenin C 28-O-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl ester (1), 3-O-β-D-glucopyranosyl-(1 → 2)-β-D-glucopyranosyluncargenin C 28-O-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl ester (2), 3-O-β-D-glucopyranosyl-(1 → 2)-β-D-glucopyranosyluncargenin C 28-O-β-D-glucopyranosyl-(1 → 4)-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl ester (3), 3-O-β-D-glucopyranosyl-(1 → 2)-β-D-glucopyranosylhederagenin 28-O-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl ester (4), 3-O-β-D-glucopyranosylarjunolic acid 28-O-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl ester (5), 3-O-β-D-glucopyranosyl-(1 → 2)-β- D-glucopyranosyl-6β,23-dihydroxyursolic acid 28-O-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl ester (6), 3-O-β-D-glucopyranosyl-6β,23-dihydroxyursolic acid 28-O-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl ester (7), asiatic acid 28-O-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl ester (8), and 3-O-β-D-glucopyranosylasiatic acid 28-O-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl ester (9), through infrared, high-resolution electrospray ionization mass spectrometry, one- and two-dimensional nuclear magnetic resonance spectral analyses. The isolates inhibited nitric oxide production in lipopolysaccharide-activated RAW 264.7 cells, with half-maximal inhibitory concentration (IC50) values of 18.8-58.5µM, compared to the positive control compound, dexamethasone, which exhibited an IC50 of 14.1µM.

Immunomodulatory and anti-inflammatory effects of bioactive compounds of Cassia angustifolia Vahl. leaf extract

Cassia angustifolia Vahl. is a medicinal plant known for its efficacy in treating various, including respiratory conditions and skin inflammation. It possesses antibacterial and anticancer properties. This work investigated the immunomodulatory and anti-inflammatory effects of C. angustifolia. The ethanol leaf extract of C. angustifolia was utilized to examine gene expression related to angiogenesis cytokines in RAW 264.7 macrophage cells. The results demonstrated a significant increase in the viability of treated macrophage RAW 264.7 cells, accompanied by an improvement in angiogenesis cytokines expression and a dose-dependent inhibition of nitric oxide production. GC-MS analysis identified 11 active components within the extract, each exhibiting distinct biological activities such as antioxidant, antitumor and anti-inflammatory effects. Notable compounds include hexadecanoic acid, 2-pentadecanone, phthalic acid, oxalic acid, carbonic acid, tricosane, undecanal and many others. In conclusion, ethanol leaf extract of C. angustifolia exhibits immunomodulatory and anti-inflammatory effects by inhibiting nitric oxide production and enhancing the expression of angiogenesis cytokines.

Discovery of structurally important cycloartane-type triterpenes from Dysoxylum binectariferum leaves and their anti-inflammatory activity

Seven undescribed cycloartane-type triterpenes (4–10) were isolated from the leaves of Dysoxylum binectariferum. The chemical structures and relative stereochemistry of all cycloartane triterpenes were determined by analysis of 1D and 2D NMR and ESI-HRMS spectroscopic data. Compound 4 was found to be a diastereoisomer of the previously discovered compound 2 (beddomeilactol). The compound 5 was identified as the methyl ester of compound 1 (beddomeilactone). The compounds 6 and 7 were found to be diastereoisomers with only a stereochemical change at C-4 and their absolute stereochemistry established by X-ray analysis. Compounds 6–8 and 10 were identified as 3,4-seco-cycloartane type triterpenes, while compound 9 was elucidated as nor-3,4-seco-cycloartane triterpene. Nitric oxide (NO) inhibition was used to assess the anti-inflammatory effects of all newly isolated natural products in LPS-induced RAW 264.7 cell line. It was found that compounds 1, 2, 6 and 8 showed good IC50 values of 14.98 ± 4.06, 38.50 ± 2.88, 36.95 ± 3.24, and 35.37 ± 4.51 µM respectively for NO production inhibition and were non-toxic to RAW 264.7 macrophages.

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.

Inhibition of NO production in LPS-activated RAW 264.7 cells of stilbenes isolated from Gnetum latifolium

The phytochemical investigation of Gnetum latifolium var. funiculare Markgr. resulted in the isolation of five compounds (1-5). Their structures were determined as bisisorhapotingenin A (1), genetifolin E (2), genetifolin K (3), piceid (4), and isorhapontin (5) based on extensive analyses of HR-ESI-MS, 1D and 2D NMR spectra. All of these five isolated compounds were evaluated for their inhibition effect on lipopolysaccharide (LPS)- induced nitric oxide (NO) production in RAW264.7 macrophages. Among them, two compounds (1 and 3) showed moderate effect on NO production inhibition in LPS-activated RAW264.7 cells with the IC50 values of 53.80 and 93.94 μM, respectively.

In vitro immunomodulation activity of protein hydrolysate from spirulina (Arthrospira platensis): the ingredient of future foods

Microalgae, especially spirulina, have been globally used as a food supplement due to their rich protein content, safety for human consumption, and provision of enhanced immunomodulatory capabilities. There are, however, few reports that have investigated the immunomodulatory properties of spirulina protein hydrolysate. Consequently, this study aims to optimize the best extraction techniques for spirulina protein hydrolysate and characterize its antioxidant activities and immunomodulation properties in vitro. The results indicated that protein hydrolysate with Flavourzyme and alkaline extraction after ultrasonication and pre-enzymatic assistant with cellulase exhibited superior antioxidant properties compared to other methods. Additionally, all the protein extracts demonstrated a dose-dependent inhibition of nitric oxide production without significantly impacting cell viability. Furthermore, in vitro immunomodulatory properties were evaluated using Candida albicans (DMST 5815) as the test pathogen, with phagocytic activity and index measurements conducted. Notably, the results correlated with the previous assessments, wherein the protein hydrolysate with Flavourzyme displayed the highest phagocytic percentage, measuring 52.3% at a concentration of 10 mg/mL. These findings suggest that enzymatically derived protein hydrolysates from spirulina could serve as a potential source for enhancing immunostimulant activity. Thus, they hold promise as natural bioactive ingredients for therapeutic purposes and the development of functional foods.

Enhanced Anti-Inflammatory and Non-Alcoholic Fatty Liver Disease (NAFLD) Improvement Effects of Bacillus subtilis-Fermented Fagopyrum tataricum Gaertner

In this study, we investigated the enhanced anti-inflammatory activity and the effects on non-alcoholic fatty liver disease (NAFLD) of fermented Fagopyrum tataricum (F. tataricum) Gaertner extract (FFT) through in vitro analysis. We utilized high-performance liquid chromatography (HPLC) to analyze the non-fermented F. tataricum Gaertner extract (NFT) and the marker components, rutin and quercetin in FFT, to confirm changes in composition due to fermentation. The anti-inflammatory activity of NFT and FFT was evaluated using a lipopolysaccharide (LPS)-induced RAW 264.7 cell inflammation model. Simultaneously, the NAFLD improvement effects were measured by evaluating lipid accumulation and the expression of lipid synthesis regulators in free fatty acid (FFA)-induced HepG2 cells. HPLC analysis confirmed an increase in rutin content after the fermentation of F. tataricum Gaertner. Upon treatment with NFT and FFT at a concentration of 400 μg/mL, LPS-induced nitric oxide (NO) production values in RAW 264.7 cells were reduced to 16.12 μM and 2.09 μM, respectively, indicating enhanced significant inhibition (p < 0.05) of NO production through fermentation. FFT demonstrated the significant inhibition (p < 0.05) of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) protein, and inflammatory cytokine mRNA expression through the nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) pathways in LPS-induced RAW 264.7 cells. In FFA-induced HepG2 cells, FFT significant suppressed (p < 0.05) lipid accumulation and the expression of sterol regulatory element binding protein (SREBP)-1c, CCAAT/enhancer binding protein (C/EBP)α proteins, and acetyl-CoA carboxylase (ACC) mRNA. The results of this study suggest the potential utilization of FFT as a material for improving NAFLD.

Role of neutrophil myeloperoxidase in the development and progression of high-altitude pulmonary edema

BackgroundNeutrophil infiltration and hypoxic pulmonary vasoconstriction induced by hypobaric hypoxic stress are vital in high-altitude pulmonary edema (HAPE). Myeloperoxidase (MPO), an important enzyme in neutrophils, is associated with inflammation and oxidative stress and is also involved in the regulation of nitric oxide synthase (NOS), an enzyme that catalyzes the production of the vasodilatory factor nitric oxide (NO). However, the role of neutrophil MPO in HAPE's progression is still uncertain. Therefore, we hypothesize that MPO is involved in the development of HAPE via NOS. MethodsIn Xining, China (altitude: 2260 m), C57BL/6 N wild-type and mpo−/− mice served as normoxic controls, while a hypobaric chamber simulated 7000 m altitude for hypoxia. L-NAME, a nitric oxide synthase (NOS) inhibitor to inhibit NO production, was the experimental drug, and D-NAME, without NOS inhibitory effects, was the control. After measuring pulmonary artery pressure (PAP), samples were collected and analyzed for blood neutrophils, oxidative stress, inflammation, vasoactive substances, pulmonary alveolar-capillary barrier permeability, and lung tissue morphology. ResultsWild-type mice's lung injury scores, permeability, and neutrophil counts rose at 24 and 48 h of hypoxia exposure. Under hypoxia, PAP increased from 12.89 ± 1.51 mmHg under normoxia to 20.62 ± 3.33 mmHg significantly in wild-type mice and from 13.24 ± 0.79 mmHg to 16.50 ± 2.07 mmHg in mpo−/− mice. Consistent with PAP, inducible NOS activity, lung permeability, lung injury scores, oxidative stress response, and inflammation showed more significant increases in wild-type mice than in mpo−/− mice. Additionally, endothelial NOS activity and NO levels decreased more pronouncedly in wild-type mice than in mpo−/− mice. NOS inhibition during hypoxia led to more significant increases in PAP, permeability, and lung injury scores compared to the drug control group, especially in wild-type mice. ConclusionMPO knockout reduces oxidative stress and inflammation to preserve alveolar-capillary barrier permeability and limits the decline in endothelial NOS activity to reduce PAP elevation during hypoxia. MPO inhibition emerges as a prospective therapeutic strategy for HAPE, offering avenues for precise interventions.

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.

3D and 2D-QSAR Studies on Natural Flavonoids for Nitric Oxide Production Inhibitory Activity

Background: Nitric oxide (NO), an important second messenger molecule, regulates numerous physiological responses, while excessive NO generates negative effects on the circulatory, nervous and immune systems. Recently, some natural flavonoids were reported to possess the capability of inhibiting LPS-induced NO production. To fully understand the nature of their own NO inhibitory activity, it is necessary to address the structural requirements of flavonoids as NO inhibitors. Objective: The objective of this work was to develop efficient QSAR models for predicting the NOinhibitory activity of new flavonoids and improving insights into the critical properties of the chemical structures that were required for the ideal NO production inhibitory activities. Methods: To provide insights into the structural basis of flavonoids as NO inhibitors, 3D quantitative structure-activity relationship (3D-QSAR) and 2D-QSAR models were developed on a dataset of 55 flavonoids using comparative molecular field analysis (CoMFA), comparative molecular similarity indices analysis (CoMSIA) and hologram quantitative structure-activity relationship (HQSAR) approaches. method: CoMFA, CoMSIA combining HQSAR methods were employed on a series of flavonoids to generate 3D and 2D-QSAR models. Results: The statistically significant models for CoMFA, CoMSIA and HQSAR resulted in crossvalidated coefficient (q2) values of 0.523, 0.572 and 0.639, non-cross-validated coefficient (r2) values of 0.793, 0.828 and 0.852, respectively. The robustness of these models was further affirmed using a test set of 18 compounds, which resulted in predictive correlation coefficients (r2 pred) of 0.968, 0.954 and 0.906. Furthermore, the models-derived contour maps were appraised for activity trends for the molecules analyzed. result: Comparative Molecular Field Analysis (CoMFA), Comparative Molecular Similarity Indices Analysis (CoMSIA) combining hologram quantitative structure-activity relationship (HQSAR) methods were employed on a series of flavonoids to generate 3D and 2D-QSAR models. Result: The obtained models can be used to predict the activities of new flavonoids and identify the key structural features affecting the NO inhibitory activities. Conclusion: The 3D and 2D-QSAR models constructed in this paper were efficient in estimating the NO inhibitory activities of flavonoids and facilitating the design of flavonoid-derived NO production inhibitors. other: none

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.

Comprehensive In Silico Analysis of Uncaria Tomentosa Extract: Chemical Profiling, Antioxidant Assessment, and CLASP Protein Interaction for Drug Design in Neurodegenerative Diseases.

Uncaria tomentosa is a traditional medicinal herb renowned for its anti-inflammatory, antioxidant, and immune-enhancing properties. In the realm of neurodegenerative diseases (NDDS), CLASP proteins, responsible for regulating microtubule dynamics in neurons, have emerged as critical players. Dysregulation of CLASP proteins is associated with NDDS, such as Alzheimer's, Parkinson's, and Huntington's diseases. Consequently, comprehending the role of CLASP proteins in NDDS holds promise for the development of innovative therapeutic interventions. The objectives of the research were to identify phytoconstituents in the hydroalcoholic extract of Uncaria tomentosa (HEUT), to evaluate its antioxidant potential through in vitro free radical scavenging assays and to explore its potential interaction with CLASP using in silico molecular docking studies. HPLC and LC-MS techniques were used to identify and quantify phytochemicals in HEUT. The antioxidant potential was assessed through DPPH, ferric reducing antioxidant power (FRAP), nitric oxide (NO) and superoxide (SO) free radical scavenging methods. Interactions between conventional quinovic acid, chlorogenic acid, epicatechin, corynoxeine, rhynchophylline and syringic acid and CLASP were studied through in silico molecular docking using Auto Dock 4.2. The HEUT extract demonstrated the highest concentration of quinovic acid derivatives. HEUT exhibited strong free radical-scavenging activity with IC50 values of 0.113 μg/ml (DPPH) and 9.51 μM (FRAP). It also suppressed NO production by 47.1 ± 0.37% at 40 μg/ml and inhibited 77.3 ± 0.69% of SO generation. Additionally, molecular docking revealed the potential interaction of quinovic acid with CLASP for NDDS. The strong antioxidant potential of HEUT and the interaction of quinovic acid with CLASP protein suggest a promising role in treating NDDS linked to CLASP protein dysregulation.

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.