Nitric Oxide Inhibition Research Articles

Anti-inflammatory, antioxidant, and anti-apoptotic properties of Thunbergia laurifolia Lindl. aqueous leaf extract on LPS-stimulated RAW 264.7 macrophages

Thunbergia laurifolia Lindl., a Thai medicinal herb, has garnered substantial attention among researchers due to its recognized efficacy as an antidote, antioxidant, and anti-inflammatory agent. Despite its extensive investigation, the specific role of T. laurifolia in apoptotic inhibition remains unexplored. Therefore, this study aimed to explore the protective properties of T. laurifolia leaf extract against lipopolysaccharide (LPS)-induced apoptosis in RAW 264.7 macrophages and determine the extractʼs total phenolic as well as flavonoid content, and its antioxidant. Moreover, the cytotoxicity including anti-inflammatory potential on LPS-stimulated RAW 264.7 macrophages were assessed by measuring the inhibition of nitric oxide (NO). The outcomes of this investigation revealed remarkable similarity between the total phenolic and flavonoid contents and antioxidant capacity of the leaf extract under scrutiny and those documented in prior studies. In addition, the extract demonstrated negligible cytotoxicity at low concentrations. Furthermore, it effectively impeded the LPS-mediated production of NO, inflammatory mediator, in the macrophages. Moreover, pre-treatment with T. laurifolia leaf extract improved LPS-induced apoptosis by reducing rate of apoptotic cell death up to 41 % at the same concentration (1.68 μL/mL) in LPS-stimulated RAW 264.7 macrophages. Collectively, these findings provide compelling initial evidence attesting to the ability of T. laurifolia leaf extract to inhibit LPS-induced apoptosis in RAW 264.7 macrophages, thereby augmenting our understanding of its potential therapeutic applications in inflammatory conditions.

NO inhibitory constituents from Glycosmis craibii var. glabra

Six novel compounds, comprising three quinolones (1a, 1b, and 2) and three flavanones (3–5), along with seven known analogs (6–13), were isolated from the 95% EtOH extract of the stems and leaves of Glycosmis craibii var. glabra. The structures of the new compounds were elucidated using HR-ESI-MS, UV, and 1D and 2D nuclear magnetic resonance (NMR) data analysis. The absolute configurations were determined through Mosher ester and electronic circular dichroism (ECD) spectral analysis. Compounds 2, 6, 9, and 10 demonstrated inhibition of nitric oxide (NO) production stimulated by lipopolysaccharide in BV-2 microglial cells, with IC50 values ranging from 13.5 to 20.1 μmol·L−1, comparable to the positive control, dexamethasone.

Resveratrol and nitric oxide synergistically enhance resistance against B. cinerea in tomato fruit by regulating phytohormones

Resveratrol (RVT), a plant antitoxin, plays an important role in plant resistance against pathogens. While nitric oxide (NO) as an essential signaling factor in disease resistance enhancement is well documented, the potential molecular interplay RVT and NO in postharvest tomato fruit defense against Botrytis cinerea (B.cinerea) still needs exploration. In this study, exogenous RVT reduced gray mold caused by B.cinerea in tomato fruit, with 20 μM being the most effective. Tomato fruit were treated with 20 μM RVT enhanced resistance against B.cinerea, as indicated by reduced symptoms of disease and improved activity of disease resistance related enzymes (PAL, PPO and CHI). In addition, RVT treatment improved the expression of SlPR1, SlLoxd and SlMYC2, and promoted the accumulation of plant hormone IAA and ABA, but reduced the expression of SlNPR1 and the level of GA3. More importantly, the combined treatment of NO donor (SNP) and RVT notably enhanced disease resistance compared to RVT or SNP single treatment. However, the combination of NO inhibitor (L-NNA) and RVT treatment even reduced the positive effect of RVT. Meanwhile, the expression of SlPR1, SlLoxd and SlMYC2 and the accumulation of IAA and ABA in RVT + SNP treated fruit were higher than those in the RVT or SNP single treatment. Thus, our data demonstrate that RVT and NO synergistically enhance resistance against B. cinerea in tomato fruit by regulating phytohormones.

Elevated plasma level of arginine and its metabolites at labor among women with preeclampsia: A prospective cohort study.

Preeclampsia complicates 3-5% of all pregnancies and is associated with higher levels of asymmetric (ADMA) and symmetric (SDMA) dimethylarginines. Dimethylarginines are inhibitors of nitric oxide, which is a uterine smooth muscles relaxant. Women with hypertensive disorders experience a shorter labor duration compared to normotensive women. However, very little is known about the possible biochemical mechanisms behind differences in labor duration. In this study we aimed to investigate if women with preeclampsia had higher levels of arginines (ADMA, SDMA and L-arginine) at labor than controls, and also investigate the association between arginines and labor duration. The study was based on data from the Swedish, Uppsala County population-based, prospective cohort BASIC, between 2009 and 2018. Arginines were analyzed by Ultra-High Performance Liquid Chromatography using plasma samples taken at labor from women with preeclampsia (n=47) and normotensive pregnancy (n=90). We also analyzed inflammation markers CRP, TNF-R1, TNF-R2 and GDF-15. Women with preeclampsia had higher levels of ADMA (p<0.001), SDMA (p<0.001), L-arginine (p<0.001), TNF-R1 (p<0.001), TNF-R2 (p=0.03) and GDF-15 (p<0.01) compared to controls. Further, ADMA and SDMA, not inflammation markers, were negatively correlated to labor duration in preeclampsia. No correlations were observed when comparing arginines and inflammation markers. Among women with preeclampsia, our novel findings of higher level of arigines, negative correlation of arginines to duration of labor and absence of correlation of arginines to inflammation markers might support the theory that it is not inflammation but arginines which could be associated with shorter duration of labor in preeclampsia.

Anti-Inflammatory Activity of Labdane and Norlabdane Diterpenoids from Leonurus sibiricus Related to Modulation of MAPKs Signaling Pathway.

Leonurus sibiricus, a widely cultivated herbaceous plant in Asian countries, exhibits diverse medicinal applications. Recent studies emphasize its pharmacological properties and efficacy in promoting bone health. Besides the known compounds and their pharmacological activities, in this study, we isolated and elucidated two new labdane-type diterpenoids, (3R,5R,6S,10S)-3,6-dihydroxy-15-ethoxy-7-oxolabden-8(9),13(14)-dien-15,16-olide (1) and (4R,5R,10S)-18-hydroxy-14,15-bisnorlabda-8-en-7,13-dione (2), a new natural phenolic compound, and a known compound from L. sibiricus using advanced spectroscopic techniques, including circular dichroism spectroscopy, high-resolution mass spectrometry, and 1- and 2-dimensional NMR. Among these, compound 1 demonstrated potent inhibition of nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) mRNA expression levels, followed by compound 2. Whereas compounds 3 and 4 do not exhibit effectiveness in RAW264.7 macrophages. Moreover, compound 1 suppressed pro-inflammatory markers induced by lipopolysaccharide (LPS) stimulation. Compound 1 also suppressed iNOS and cyclooxygenase-2 (COX-2) protein levels and downregulated pro-inflammatory cytokines. Additionally, compound 1 showed inhibition of the phosphorylation of p38, JNK, and ERK, key mediators of the MAPK signaling pathway. These findings indicate that a natural-derived product, compound 1, might be a potential candidate as an anti-inflammation mediator.

Development of Bacillus stratosphericus Lysate Concentrate to Control Sebum Secretion through In vitro Studies and Clinical Trial

The sebum on human skin is generated for various causes. The composition of the formed sebum increases the proliferation of Cutibacterium acnes (C. acnes) residing on the skin. As C. acnes proliferates, it produces skin irritants that stimulate the sebaceous glands, increasing sebum production. Skin troubles such as acne may occur. The lysate concentrates of Bacillus stratosphericus (B. stratosphericus), first discovered in the stratosphere, confirmed a 66.35% inhibition of Nitric Oxide (NO) production at 0.50 mg/ml concentration in vitro. Additionally, the growth inhibition efficacy of B. stratosphericus lysate concentrate (BSLC) against C. acnes was confirmed, showing a 95.1% inhibition of growth proliferation at a consistency of 0.50 mg/ml. Based on the in vitro results, the efficacy of BSLC in degrading and reducing sebum was confirmed by reacting it with artificial sebum to various concentrations. The results showed a concentration-dependent decrease in artificial sebum ccording to the efficacy results confirmed in vitro, a clinical trial was conducted to evaluate the daily sebum reduction efficacy of a serum formulation containing 50 mg/ml of BSLC. After a 4-week application, the test group containing BSLC determined a significant 28.68% reduction in sebum levels, demonstrating the practical implications of the research. In conclusion, BSLC is considered to have sufficient industrial value as a valuable ingredient for the cosmetics industry aimed at sebum improvement.

Bio-guided isolation of anti-inflammatory compounds from Tilia tuan Szyszyl. flowers via in vitro and in silico study

Ethnopharmacological relevanceTilia tuan Szyszyl. is a perennial arboreal plant renowned for its medicinal and economic significance. All the roots, stems, leaves, flowers, and fruits have been used medicinally by the folk Bai and Yi people in Yunnan province, China, to treat inflammation, rheumatism and pain for a long time. The detailed chemical constituents and their anti-inflammatory mechanisms remain unexplored. Aim of the studyThe objective of this study was to investigate the main anti-inflammatory constituents of T. tuan flowers through bio-guided isolation and to evaluate their anti-inflammatory effects relevant to its traditional medicinal use. Material and methodsBio-guided isolation was conducted on the extract of T. tuan flowers using a combination of column chromatography and preparative HPLC. The structures were established by a combination of extensive spectroscopic analyses and single-crystal X-ray diffraction analysis. The anti-inflammatory activity in vitro was evaluated by measuring the inhibition of nitric oxide (NO) production in LPS-stimulated RAW 264.7 cells. TNF-α and IL-6 levels were evaluated by ELISA. The expression of COX-2 and NF-κB was assayed via western blotting, and in-silico molecular docking was conducted to explore the potential mechanism. ResultsTwenty-two compounds, including ten flavonoids (1−10), seven phenylpropanoids (11−17), three triterpenoids (19−21), one sterol glucoside 18, and one glyceride 22, were identified from T. tuan flowers for the first time. Among them, 1 is a new compound. It is noted that 1, 5, 7, 8, 10, 17, and 22 exhibited remarkable anti-inflammatory activity against NO production with a range of 5.2–34.5 μM, superior to the positive control L-NMMA. Moreover, the new compound 1 inhibited significantly inflammatory factors IL-6 and TNF-α. ConclusionThe results show that flavonoids including the new one and the phenyl-propanoid are the primary active constituents of T. tuan flowers, responsible for its ethno-pharmacological uses on osteoarthritis and rheumatism. T. tuan flowers could be a promising therapeutic agent to modulate inflammatory diseases.

Phenolic Bisabolane Sesquiterpene Derivatives from an Arctic Marine-derived Fungus Aspergillus sydowii MNP-2

Background:: Filamentous fungi in the genus Aspergillus are well known for their important roles in production of bioactive secondary metabolites with diversely chemical structures and potential application in pharmaceutical industry. Objective:: The present study aimed to investigate the phenolic bisabolane sesquiterpene (PBS) derivatives from an Arctic marine-derived fungus Aspergillus sydowii MNP-2. Methods:: In this study, antimicrobial activities were carried out according to the broth microdilution assay, nitric oxide (NO) production in mouse macrophages (RAW264.7) and BV2 microglial cells was used to detect the inhibitory effect of compounds in inflammatory reactions, and in vitro inhibitory cell proliferation activity was determined by the cell counting kit-8 (CCK-8) assay. Results:: In this work, chemical investigation of an Arctic marine-derived strain A. sydowii MNP-2 led to the isolation of 11 PBSs (1-11) using various chromatographic methods. Their chemical structures were unambiguously determined by 1H NMR spectroscopy and mass spectrometry analyses as well as comparison with literature data. It is noteworthy that compounds 1, 7 and 11 were firstly obtained from A. sydowii. Antimicrobial assay showed that these chemicals had no potent inhibitory effect on Staphylococcus aureus, Escherichia coli, and Candida albicans with MIC values &gt; 16 μg/mL. Additionally, the inhibition of nitric oxide (NO) production in lipopolysaccharide (LPS)- induced inflammation in mouse macrophages (RAW264.7) and BV2 microglial cells were all below 10% for compounds 4-6 and 8, indicating almost negligible anti-inflammatory efficacy. Among the tested compounds 4-6 and 8 for tumor-cell proliferation inhibition activities, compound 5 demonstrated the strongest inhibitory effect against human acute promyelocytic leukemia cells (HL-6) with a 44.76% inhibition rate. Conclusion:: In the present study, 11 PBS derivatives were purified and characterized from the solidand liquid-state fermentations of the Arctic marine-derived fungus A. sydowii MNP-2. Unfortunately, none of these metabolites had significant antimicrobial, anti-inflammatory, or tumor-cell proliferation inhibition activities.

Evaluation of antioxidant and anti-inflammatory properties, bioactive compound profiling, and molecular mechanisms of a multicomponent Thai herbal formulation

BackgroundMedicinal plants have long been used as dietary supplements to reduce the risk of chronic diseases, inflammation, cancer, and metabolic syndromes. Traditional remedies possess therapeutic properties, making them promising candidates for further investigation and potential pharmaceutical development. Aim of the studyThis study aimed to evaluate the efficacy of Belog Plus (BP), a polyherbal product containing five Thai medicinal plants: Citrus aurantifolia, Cannabis sativa, Tiliacora triandra, Alpinia galanga, and Piper nigrum, combined with safflower seed oil. This study focused on quantifying the total phenolic content (TPC) and total flavonoid content (TFC), and identifying key herbal biomarkers that contribute to its therapeutic properties. In addition to assessing the antioxidant and anti-inflammatory potential of the plant extracts, the study explored the molecular mechanisms underlying their anti-inflammatory activity, offering a comprehensive understanding of BP's efficacy. MethodsThe ethanolic extract of BP and its components were assessed for antioxidant potential using DPPH, FRAP, and H2O2 induced oxidative stress. The anti-inflammatory activity was evaluated using the Griess reaction assay, and the mechanisms were explored through mRNA and protein expression analysis. The TPC and TFC were measured using the Folin–Ciocalteu (F–C) assay and the aluminium chloride colorimetric assay, respectively, while herbal biomarkers were identified using LC-MS/MS analysis. ResultsThe results revealed that the ethanolic extract of BP (6.25-100 µg/ml) exhibited a potent anti-inflammatory effect through nitric oxide inhibition (IC50 24.4 μg/ml) and significantly reducing the expression of inflammatory genes iNOS, COX-2, IL-6, and TNF-α. Additionally, it significantly reduced the expression of inflammatory proteins iNOS and COX-2 at concentrations of 6.25-25 µg/ml. Among the individual components, C. aurantifolia, A. galanga, and P. nigrum showed potent anti-inflammatory effects with IC50 values of 35.7, 6.5, and 23.3 μg/ml, respectively. The TPC and TFC results demonstrated significant levels of phenolic (80.6 µg GAE/mg) and flavonoid (47.4 µg catechin/mg) compounds. Additionally, the LC-MS/MS analysis identified a variety of bioactive compounds known for their anti-inflammatory properties. ConclusionThese findings support BP's potential use as a dietary supplement to mitigate the risk of chronic inflammatory diseases, with its ethanolic extract containing bioactive phenolic and flavonoid compounds that significantly suppress iNOS and COX-2 protein expressions.

Synthesis, Characterization and Targeted Drug Delivery of Curcumin-Loaded PLGA Nanoparticles

Background: Nanoparticle-based drug delivery systems have revolutionized therapeutic delivery, offering advantages such as enhanced bioavailability, controlled release, and targeted delivery to diseased tissues. Among these, polymeric nanoparticles, especially those utilizing poly(lactic-co-glycolic acid) (PLGA), have shown significant potential due to their biocompatibility, biodegradability, and stability. Curcumin, a polyphenolic compound with potent anti-inflammatory and anticancer properties, faces clinical limitations due to poor solubility and low bioavailability. Encapsulation in PLGA nanoparticles could enhance curcumin’s therapeutic efficacy by improving stability, controlled release, and targeted delivery. Methods: Curcumin-loaded PLGA nanoparticles were synthesized using a solvent evaporation method. Nanoparticles were characterized using dynamic light scattering (DLS) for size and zeta potential, and scanning electron microscopy (SEM) for morphology. In vitro drug release was assessed using a dialysis method, while cytotoxicity and anti-inflammatory activity were evaluated using MTT and nitric oxide inhibition assays, respectively. Results: DLS analysis revealed an average particle size of 150 ± 10 nm and a zeta potential of -25 ± 2 mV, indicating stability and suitability for therapeutic applications. SEM imaging showed smooth, spherical nanoparticles. Drug release studies displayed a biphasic pattern, with an initial burst followed by sustained release over 72 hours. Cytotoxicity assays demonstrated enhanced anticancer activity of curcumin-loaded nanoparticles compared to free curcumin, with a significantly lower IC50 value (5 ± 0.5 µM versus 15 ± 1.0 µM). Anti-inflammatory studies showed a 60% inhibition of nitric oxide production in LPS-induced macrophages, indicating improved anti-inflammatory efficacy over free curcumin. Conclusion: Curcumin-loaded PLGA nanoparticles show promise as a targeted drug delivery system, with enhanced stability, controlled release, and increased therapeutic efficacy. These findings support further in vivo studies to validate the clinical potential of curcumin nanoparticles in cancer and inflammatory diseases, providing a foundation for advanced therapeutic applications.

Development of Nitric Oxide-Donating Netarsudil Derivatives as a Synergistic Therapy for Glaucoma with Reduced Ocular Irritation.

Based on the synergistic therapeutic effect of nitric oxide (NO) and Rho-associated protein kinase (ROCK) inhibitors on glaucoma, a series of NO-donating Netarsudil derivatives were designed, synthesized, and their activities in vitro and in vivo were evaluated. Among them, (S)-10e released an appropriate amount of NO in aqueous humor in vitro and displayed potent ROCK inhibition. Topical administration of (S)-10e significantly lowered intraocular pressure in an acute ocular hypertension rabbit model and protected retinal ganglion cells in a magnetic microbead occlusion mouse model. A metabolism investigation revealed that (S)-10e released 7a, a metabolite after NO releasing, and 13, an active metabolite of (S)-Netarsudil, in rabbit eyes. Notably, introducing an NO donor moiety attenuated ROCK inhibition-induced ocular irritation in an sGC-independent manner, suggesting that the attenuated conjunctival hyperemia effect of (S)-10e is related to the NO-induced protein S-nitrosation of phosphodiesterase 3A (PDE3A). Overall, (S)-10e is a promising candidate for glaucoma treatment.

Iron chelating, antioxidant, and anti-inflammatory properties of brazilin from Caesalpinia sappan Linn

BackgroundIron overload and inflammation are severe conditions that can lead to various chronic diseases. However, the current iron chelator drugs have their limitations. The phytochemical compounds from herbals, such as brazilin, the major active compound in Caesalpinia sappan Linn., have significant therapeutic potential in various chronic diseases. Our study was designed to examine the effect of brazilin on iron chelating properties, antioxidant activity in hepatocytes, and anti-inflammatory potential in macrophages. MethodsThis study focused on the isolation, purification, and evaluation of brazilin, the principal bioactive constituent found in C. sappan wood. Brazilin was extracted via methanol maceration followed by column chromatography purification. The purified compound was characterized using high-performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR) spectroscopy, and mass spectrometry (MS). The antioxidant potential of brazilin was assessed by in vitro assays, including 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azinobis-(3-ethylbenzthiazolin-6-sulfonic acid (ABTS), and ferric-reducing antioxidant power (FRAP). Furthermore, its cellular antioxidant activity was evaluated using hydrogen peroxide-induced oxidative stress in the hepatocellular carcinoma cell line (Huh-7). The iron-chelating capacity of brazilin was determined spectrophotometrically, and Job's plot method was used to elucidated the stoichiometry of the iron-brazilin complex formation. The anti-inflammatory properties of brazilin were investigated in lipopolysaccharide (LPS)-stimulated macrophages (RAW 264.7). Nitric oxide (NO) inhibition was quantified using the Griess reagent, while the expression levels of pro-inflammatory cytokines, interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α), were evaluated by RT-qPCR. ResultsThe results demonstrated that brazilin exhibited potent antioxidant activity in vitro and hepatocytes in a concentration-dependent manner. It also showed anti-inflammatory activity, in which NO production was significantly reduced and IL-6 and TNF-α expression in LPS-induced macrophages were repressed. Furthermore, it can bind ferric and ferrous ions. Brazilin acts as a bidentate iron chelator that forms a complex with iron in a 2:1 ratio, and two water molecules are used as additional chelators in this complex. ConclusionsOur findings have significant implications. Brazilin can potentially alleviate the harmful effects of iron-induced oxidative stress and inflammatory disorders.

Effects of L-NAME and air exposure on mitochondrial energetic markers, thyroid hormone receptor/regulator system and stress/ease-responsive receptor expression in the brain/gut axis of zebrafish

As a signal molecule, nitric oxide (NO) has several physiological actions in fish. However, the action of NO on the brain/gut axis, a classic inter-organal axis that bridges the gastrointestinal tract and the CNS, still requires more understanding. The short-term in vivo action of a NO inhibitor, N-omega-nitro-L-arginine methyl ester (L-NAME), on mitochondrial energetic markers and the receptor expression of thyroid hormone (TH) and neuroendocrine hormones involved in stress/ease response was tested in the brain/gut axis of zebrafish exposed to either in non-stressed or air-exposed condition. L-NAME treatment decreased the NO content in brain and gut segments in non-stressed fish but rose upon L-NAME treatment in air-exposed fish that corresponded with the activation of inos, nnos, hif1a and hif1an transcript expressions. The brain/gut segments that showed spatial and differential sensitivity to L-NAME, modified the transcript expression patterns of stress (adra2da, adrb1, nr3c2)- and ease-responsive (htr2b, slc6a4a, mtnr1aa) hormone receptors. The expression pattern of the TH receptor/regulator system (thra, thrb, dio1, dio2, dio3) becomes more active in gut segments than brain segments upon L-NAME challenge in stressed zebrafish. The data provide evidence for a novel role of NO as an integrator of brain/gut axis segments in zebrafish, where the endogenously produced NO in mid-brain/posterior-gut axis aligns together upon air-exposure stress, providing a lead role to the posterior gut that activates and directs the neuroendocrine receptor expressions of stress/ease responsive genes. The data further invites studies exploring the therapeutic potential of L-NAME in this biomedical model to control the brain/gut axis segments.

Immunomodulatory Effects and Regulatory Mechanisms of (R)-6-HITC, an Isothiocyanate from Wasabi (Eutrema japonicum), in an Ex Vivo Mouse Model of LPS-Induced Inflammation.

The present study aimed to investigate the effects of (R)-(-)-1-isothiocyanato-6-(methylsulfinyl)-hexane [(R)-6-HITC], the major isothiocyanate present in wasabi, in an ex vivo model of inflammation using lipopolysaccharide-stimulated murine peritoneal macrophages. (R)-6-HITC improved the immune response and mitigated oxidative stress, which involved suppression of reactive oxygen species, nitric oxide, and pro-inflammatory cytokines (IL-1β, IL-6, IL-17, IL-18, and TNF-α) production and downregulation of pro-inflammatory enzymes such as inducible nitric oxide synthase, COX-2, and mPGES-1. In addition, (R)-6-HITC was able to activate the Nrf2/HO-1 axis while simultaneously inhibiting key signaling pathways, including JAK2/STAT3, mitogen-activated protein kinases, and canonical and noncanonical inflammasome pathways, orchestrating its potent immunomodulatory effects. Collectively, these findings demonstrate the potential of (R)-6-HITC as a promising nutraceutical for the management of immuno-inflammatory diseases and justify the need for further in vivo validation studies.

Guaijaverin And Epigallocatechin Gallate Exerts Antiinflammatory And Antiallergenic Effects Through Interleukin-12 Production.

Our aim in the current study was to determine the in vitro and invivo synergistic antiinflammatory and antiallergic effect associated with the IL-12 production of guaijaverin and epigallocatechin gallate (EGCG) complex (GEC) and ILS-F-2301 (2:8 extract of Psidium guajava and Camellia sinensis). Compared to EGCG alone, GEC showed synergistic inhibition of nitric oxide (NO), inducible NO synthase, and cyclooxygenase-2 by 3.8, 5.1, and 4.1%, respectively. The downregulation of interleukin-12 (IL-12) by 2,4-dinitrophenyl-human serum albumin conjugate/DNP-immunoglobulin E or ovalbumin (OVA) was synergistically increased by GEC by about 7.5% or 5.4% compared to EGCG alone. The level of downregulation of IL-12 in plasma increased by 100 mg/kg with ILS-F-2301 (28.7%) when compared to the OVA/Alu-treated group. Also, GEC synergistically increased by GEC by about 7.5% or 5.4% compared to EGCG alone. The level of down and cyclooxygenase C synergistically inhibited p-Akt, PI3K, mTOR, p-STAT6, and GATA3 by 4.9%, 4.1%, 19.2%, 23.8%, and 35.3%, respectively, while increasing the expressions of p-STAT1 and T-bet (showing 53.3% and 9.4% activation) when compared to EGCG alone. In an allergenic rhinitis mouse model, 100 mg/kg of ILS-F-2301 was shown to inhibit p-Akt, PI3K, mTOR, p-c-Jun N-terminal kinase (p-JNK), p-extracellular signal-regulated kinase (p-ERK), and p-p38 by 23.3%, 43.8%, 17.2%, 32.2%, 29.1%, and 41.8% when compared to the OVA/Alu-sensitized group. Taken together, our findings suggest that ILS-F-2301 may have potential as a functional food for alleviating antiallergic rhinitis.

Potent Cytotoxicity and Nitric Oxide Suppression of Compounds Derived from Kaempferia elegans Rhizomes: Molecular Modeling on EGFR Inhibition

Potent Cytotoxicity and Nitric Oxide Suppression of Compounds Derived from Kaempferia elegans Rhizomes: Molecular Modeling on EGFR Inhibition

Changes in nitric oxide inhibitors and mortality in critically ill patients: a cohort study

BackgroundOptimal balance between macro- and microcirculation in critically ill patients is crucial for ensuring optimal organ perfusion. Nitric oxide (NO) is a regulator of vascular hemostasis and tone. The availability of NO is controlled by asymmetric dimethylarginine (ADMA), symmetric dimethylarginine (SDMA), and the availability of the NO substrates arginine and homoarginine. We investigated the changes in plasma concentrations of ADMA, SDMA, arginine, and homoarginine days 1–5 of intensive care unit (ICU) admission and the association between the change in concentration days 1–3 and 30-day all-cause mortality.MethodsSingle-center cohort study of adult critically ill patients from the ICU at Copenhagen University Hospital – North Zealand. ADMA, SDMA, arginine, and homoarginine (NO-biomarkers) were measured on days 1–5. Initially, we determined the changes in NO-biomarkers days 1–5 with linear mixed models, and subsequently how the changes in NO-biomarkers days 1–3 were associated with 30-day all-cause mortality. Post-hoc we analyzed the association between plasma concentration at admission and 30-day all-cause mortality.ResultsIn total 567 out of 577 patients had plasma samples from days 1–5. Plasma concentrations of ADMA and arginine increased from days 1–5. SDMA concentrations increased from days 1–2, followed by a decrease from days 2–5. Concentrations of homoarginine did not change from days 1–3 but slightly increased from days 3–5. In total 512 patients were alive 3 days after ICU admission. Among these patients, a daily twofold increase in ADMA concentration from days 1–3 was associated with decreased mortality in multivariate analysis (HR 0.45; 95% CI 0.21–0.98; p = 0.046). An increase in SDMA, arginine, or homoarginine was not associated with mortality. Post-hoc we found that a twofold increase in ADMA or SDMA concentrations at admission was associated with mortality (HR 1.78; 95% CI 1.24–2.57; p = 0.0025, and HR 1.41; 95% CI 1.05–1.90; p = 0.024, respectively).ConclusionsIncreasing ADMA concentrations on days 1–3 are inversely associated with mortality, however not with the same strength as high ADMA or SDMA concentrations at admission. We suggest that admission concentrations are the focus of future research on ADMA and SDMA as predictors of mortality or potential therapeutical targets in ICU patients.

Molecular Dynamics-Ensemble Docking and Biophysical Studies for Structure-Based Identification of Non-Amino Acidic Ligands of DDAH-1.

Dimethylarginine dimethylaminohydrolase-1 (DDAH-1) accounts for the catabolism of the endogenous inhibitors of nitric oxide (NO) synthases, namely, ADMA (Nω,Nω-dimethyl-l-arginine) and NMMA (Nω-monomethyl-l-arginine). Inhibition of DDAH-1 may prove a therapeutic benefit in diseases associated with elevated nitric oxide (NO) levels by providing a tissue-specific increase of ADMA and NMMA. In this work, we have used molecular dynamics to generate a pool of DDAH-1 conformations in the apo and holo forms. Ensemble docking has been instrumental in screening an in-house fragment-based library of 824 compounds. Resulting virtual hits have been validated for their binding activity to recombinant human DDAH-1 using microscale thermophoresis (MST). As a key result, three non-amino acidic ligands of DDAH-1 (VIS212, VIS268, VIS726) are identified with higher binding efficiency index than ADMA. Amid these compounds, purpurogallin (VIS726) proves a potent ligand of DDAH-1, showing a mixed behavior of enzymatic inhibition in a biochemical assay. This finding widens the panel of known molecular targets of purpurogallin and provides clues into the molecular mechanisms of its cellular NO inhibition activity as well as its anti-inflammatory and neuroprotective effects.

Ent-Kaurane diterpenoids from Isodon henryi and their anti-inflammatory activities

Phytochemical investigation of the 70% ethanol extract of Isodon henryi Kudô afforded fifteen ent-kaurane diterpenoids, including nine previously undescribed compounds, named isohenolides C–K (1–9). Compounds 1–6 featured an unusual 6,7;8,15-diseco-7,20-olide ent-kaurane diterpenoid scaffold, in which 1 also possessed an 11,15-lactone ring while 2–6 all contained a free α-methylene-γ-carboxylic acid. Compound 6 was also a rare 6,8-cyclo-7,20-olide ent-kauranoid. Their structures were elucidated primarily by HRESIMS, 1D and 2D NMR spectroscopy, electronic circular dichroism and X-ray diffraction (Cu Kα) methods. Additionally, most compounds were also screened for anti-inflammatory actions against lipopolysaccharide-induced RAW 264.7 cells, and compounds 9 and 13 exhibited stronger nitric oxide inhibition, with IC50 values of 15.99 ± 0.75 and 18.19 ± 0.42 μM, respectively.

Therapeutic potential of sulfated glycosaminoglycan from seafood Asian green mussel (Perna viridis): Insights from an in vivo study

The Asian green mussel, Perna viridis, is widely consumed as seafood throughout the coastal regions of the Indo-Pacific area owing to its significant pharmaceutical and biomedical benefits. A pharmacologically active (1 → 3)/(1 → 4) linked sulfated glycosaminoglycan (PVP-2) with β-(1 → 3)-GlcNSp, and α-(1 → 4)-GlcAp units was isolated from P. viridis. PVP-2 (at 1–10 μg/mL) demonstrated a suppression of nitric oxide (NO) secretion in lipopolysaccharide-induced RAW 264.7 cells. Notably, PVP-2 at 5 μg/mL effectively restored NO levels (0.32 μg/mL) to homeostasis by downregulating excessive production (1.22 μg/mL). Furthermore, PVP-2 (at 22 mg/kg body weight) showed effective reduction of carrageenan-induced paw edema (82% inhibition at 5 h) in time dependent manner. This reduction was found to be comparable with standard treatment (87% inhibition of paw edema at 5 h). Treatment with PVP-2 (at 110 mg/kg body weight) resulted substantial inhibition of formalin-induced paw edema up to 10 days. At the end of the 10th day, the reduction in paw thickness was significantly greater (a four-fold increase) in the PVP-2 treated group compared to the formalin-induced group. Furthermore, PVP-2 displayed potent anti-inflammatory effect by attenuating cyclooxygenases (COX-1, 2) and 5-lipoxygenase (5-LOX) (IC50 < 2 mg/mL) enzymes with a higher selectivity index (IC50 COX-1/IC50 COX-2 ∼ 1.7) towards inducible pro-inflammatory COX-2. Therefore, inhibition of the NO production along with the reduction of paw edema, highlights the therapeutic potential of sulfated glycosaminoglycan from seafood Asian green mussel, as a naturally derived functional food to mitigate inflammation related disorders.