Nitric Oxide Inhibition Research Articles

Isolation, identification of endophytic fungi extracts from Codiaeum variegatum and assessment of their anti-amoebic and anti-inflammatory potential

Intestinal amoebiasis (IA) remains a global public health concern. It is often confused with an inflammatory bowel disease, and is caused by Entamoeba histolytica. Although it is more widespread in tropical countries, IA is increasingly becoming a subject of concern in so-called developed countries. The limits of conventional therapy and the concern to promote phytotherapy in a sustainable and ecological manner inspired this work, during which the anti-amoebic and anti-inflammatory potential of endophyte fungi extracts isolated from Codiaeum variegatum were evaluated. C. variegatum leaves and bark was harvested at the University of Yaounde 1. Those two fragments undergo surface decontamination then were placed in petri dishes containing PDA (Potato Agar Dextrose) supplemented with amoxicillin. The cultures obtained were then subcultured in SDA (Sabouraud Dextrose Agar) and CZA (CZapek dextrose Agar) media. The resulting growth was observed using an optical microscope with an x40 objective for grouping on the basis of macroscopic and microscopic characteristics. Screening of extracts of these fungi by the Trypan blue counting method on clinical isolates of E. histolytica, made it possible to retain 4 extracts (6, 8, 14, 18) of fungi with anti-amoebic activity. Subsequently, the anti-inflammatory potential of these extracts was evaluated on a primary culture of macrophages activated by Saccharomyces cerevisiae through the inhibition of nitric oxide (NO) production, NADPH oxidase activity, and activation of alkaline phosphatase (ALP). From that culture, 28 endophytic fungi were isolated, 13 belonging to 7 genus (Absidia sp, Alternaria sp, Aspergillus sp, Trametes sp, Trichoderma sp, Fusarium sp and Geotrichum sp). Anti-amoebic tests showed that extract 14 was the most active with an inhibitory concentration 50 (IC50) of 289.87±18.57; 207.73±22.36 and 87.79±12.51µg/mL respectively after 24, 48 and 72 hours of incubation. Chemical screening of the extracts revealed the majority presence of flavonoids in the 4 extracts (6, 8, 14, and 18). The anti-inflammatory tests of these 4 extracts revealed better anti-inflammatory activity in extract 6 (IC50= 95.94 ± 21.07 µg/mL for NO; IC50= 11.78±0.27 µg/mL for NADPH oxidase). Extract 14 had the best effect on the inhibition of NO production (IC50 = 8.56±2.13 µg/mL) and on the inhibition of the growth of trophozoites. Endophytic fungi extract from C. variegatum have demonstrated promising anti-amoebic and anti-inflammatory potential and therefore deserve special attention for the development of natural and ecological alternative treatment.

Relaxant Effect of Rosuvastatin in Isolated Rat Aorta with Perivascular Adipose Tissue

Objective: Rosuvastatin displays favorable pleiotropic effects on vascular system to reduce the risk of cardiovascular events besides providing an intensive reduction in LDL-C levels. The role of perivascular adipose tissue (PVAT) in modulating the vasorelaxant effect of rosuvastatin is not evaluated so far. The present study aimed to investigate the vascular relaxant effect of rosuvastatin in rat aortic rings with intact PVAT, as well as to evaluate the possible mechanisms underlying this effect in relation to nitric oxide (NO) and prostaglandin pathways. Methods: Thoracic aorta rings with intact PVAT, isolated from male Wistar rats (n=5), were mounted on an isolated organ bath system. Endothelium-dependent responses to acetylcholine (Ach,10-6-10-4M) were obtained in aortic rings precontracted submaximally with phenylephrine (Phe,10-6-3x105M). The concentration-dependent relaxant effect of rosuvastatin (10-7-10-4M) was examined in the absence and presence of NO inhibitor, L-NOARG (10-4M, 30min.) and cyclooxygenase inhibitor, indomethacin (10-5M, 30min.). Vascular relaxation capacity of aortic rings was checked by the nitrovasodilator, sodium nitroprusside (SNP,10-6 M) at the end of the experiments. Results: Rosuvastatin (10-7-10-4M) produced concentration-dependent relaxations in Phe-precontracted rat aortic rings with intact PVAT. Pretreatment with L-NOARG significantly attenuated the relaxant responses to rosuvastatin in isolated rat aortic rings with intact PVAT. However, pretreatment with indomethacin did not modify the relaxations to rosuvastatin. In the aortic rings, maximal relaxation responses to Ach and SNP were determined to be 75.87±2.68% and 102.54±2.92%, respectively. Conclusions: This study will provide a basis for investigating the interaction between PVAT and statins in vascular homeostasis.

Stachys byzantina K. Koch in the Treatment of Skin Inflammation: A Comprehensive Evaluation of Its Therapeutic Properties.

Stachys byzantina is a plant widely cultivated for food and medicinal purposes. Stachys species have been reported as anti-inflammatory, antibacterial, anxiolytic, and antinephritic agents. This study aimed to evaluate the anti-inflammatory potential of the ethanolic extract (EE) from the aerial parts of S. byzantina and its most promising fraction in models of acute and chronic inflammation, including a psoriasis-like mouse model. The EE was fractionated into hexane (HF), dichloromethane (DF), ethyl acetate (AF), and hydroalcoholic (HD) fractions. Screening for anti-inflammatory activity based on nitric oxide inhibition (IC50 μg/mL: HF 24.29 ± 5.87, EE 176.45 ± 18.65), hydroxyl radical scavenging (HF 3.89 ± 0.61, EE 6.38 ± 2.25), β-carotene/linoleic acid assay (HF 10.13 ± 3.81, EE 25.64 ± 2.12), and ORAC identified HF as the most active fraction. Topical application of HF effectively reduced croton oil- and phenol-induced ear edema in mice, with no statistical difference to the reference drugs. A formulation containing HF showed significant activity in the imiquimod-induced psoriasis model, reducing pro-inflammatory cytokines and nitric oxide production in macrophages, with no cytotoxicity to skin cells. Phytochemical analysis of HF revealed the presence of terpenes, steroids (491.68 ± 4.75 mg/g), phenols (34.30 ± 4.96 mg/g), flavonoids (151.77 ± 6.66 mg/g), and α-tocopherol, which was identified and quantified by HPLC-UV analysis (10.56 ± 0.97 mg/g of HF). These findings highlight the therapeutic potential of S. byzantina for skin inflammation, particularly contact dermatitis and psoriasis, encouraging further studies, including in human volunteers.

Management of hypertension and coronary artery disease in vasculitis patients using glucocorticoids

Systemic vasculitis is an autoimmune disorder characterized by inflammation and damage to blood vessels, often involving multiple organ systems. Glucocorticoids (GCs) are a cornerstone in managing vasculitis and other rheumatologic conditions due to their potent anti-inflammatory properties, but their use is associated with significant systemic side effects. This review evaluates the cardiovascular (CV) risks associated with glucocorticoid use, focusing on hypertension and atherosclerosis, and highlights strategies for minimizing these risks while managing systemic inflammation in vasculitis patients. Glucocorticoids, despite their efficacy in disease management, contribute to CV complications in a dose- and duration-dependent manner. Hypertension arises through mechanisms such as nitric oxide suppression, renin-angiotensin-aldosterone system activation, and vascular effects. Atherosclerosis is accelerated by endothelial dysfunction, oxidative stress, and inflammation, increasing risks for coronary artery disease and acute CV events. Daily doses exceeding 5 mg of prednisolone are linked to a marked rise in CV risks, even with short-term use. Moreover, systemic inflammation from vasculitis compounds these risks, making it challenging to disentangle GC side effects from the underlying disease pathology. Glucocorticoid therapy requires careful management, prioritizing the lowest effective dose and shortest duration possible. Routine CV risk assessments, blood pressure monitoring, and targeted interventions such as lifestyle modifications and pharmacologic therapies are crucial for mitigating adverse outcomes. Future studies should aim to define “safe” GC thresholds and refine treatment protocols to balance efficacy and safety in vasculitis management.

Taxifolin: A flavonoid in the freezing medium augments post-thaw semen quality and in vivo fertility potential of buffalo bull spermatozoa

This comprehensive study was carried out to investigate the effects of taxifolin in the freezing medium on post-thaw semen quality, and fertility potential of buffalo bull spermatozoa. Taxifolin was also evaluated for radical scavenging activity through DPPH (2, 2-diphenyl-1-picrylhydrazyl), and Nitric oxide (NO) inhibition (%) during in vitro conditions. Collected semen samples from four buffalo bulls were initially evaluated (consistency, volume, motility, and concentrations); the accepted samples were pooled, and diluted in extenders containing different doses of taxifolin (0 μM [control], 2 μM, 5 μM, 10 μM, and 20 μM). Diluted semen was gradually cooled (2 h, 4 °C), equilibrated (4 h, 4 °C), and then frozen in liquid nitrogen (−196 °C). Data analysis revealed that taxifolin supplementation (10 μM) displayed the highest DPPH-scavenging ability, and NO inhibition compared to the control (% DPPH, 67.31 vs. 13.50; and % NO, 78.25 vs. 21.25) respectively. Moreover, taxifolin supplementation (10 μM) significantly (P < 0.05) improved progressive motility (%), average path velocity (μm/sec), straight-line velocity (μm/sec), and seminal plasma glutathione peroxidase (μM), and superoxide dismutase (U/mL) of buffalo bull spermatozoa than control. Sperm plasma membrane integrity, mitochondrial transmembrane potential, acrosome integrity (%) and seminal plasma adenosine triphosphate (nmol/million), and total antioxidant capacity (μM/L) were enhanced significantly with taxifolin (10 and 20 μM) supplementing group. Lastly, taxifolin supplementation significantly improved the fertility rate (%, 69.09 vs. 40.38) compared to the control. Further studies to assess mechanisms by which taxifolin improves semen quality and fertility of buffalo spermatozoa are warranted.

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. 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. 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.

Evaluating the quality, physicochemical properties, and biological activities of Centauri® honey from Turkey

Honey, a natural production of honeybees, underscores Turkey's prominence as one of the world's largest honey producers. The country's diverse flora fosters optimal conditions for beekeeping, resulting in the production of exceptional-quality honey. Centauri® Honey is uniquely sourced from remote alpine regions in Turkey, characterized by bees that inhabit isolated caves with access to medicinal endemic flowers year-round, promoting a unique and pristine production environment. This study evaluates the quality, physicochemical properties, nutritional parameters, and biological activities of five types of Centauri® Honey. The in vitro studies highlighted significant nitric oxide inhibition in Centauri® Cave Honey, suggesting it is a promising candidate for further research and potential therapeutic applications, especially in enhancing immune responses by reducing the inflammatory response. This study confirms that Centauri® honey not only adheres to the rigorous standards of the Codex Alimentarius but also offers substantial health benefits, advocating for continued research into its medicinal applications.

Exploring the Anti-melanogenic, Antioxidant, and Anti-inflammatory Activities of A Composition: Glabridin, Resveratrol and Ellagic Acid

Background: Plant extracts have wide applications in food, nutrition, and cosmetics, which results in a deeper investigation of natural ingredients. Numerous natural ingredients have been demonstrated to exhibit multiple activities, including antioxidation, anti-inflammation, and anti-melanogenesis. However, their combinations have not been well investigated, which could provide stronger performance with less toxicity and easier applications. Methods: We used B16F10 cells treated with alpha-melanocyte stimulating hormone (αMSH) for melanogenesis-related studies, including cellular melanin content, tyrosinase activity, and gene or protein expression. MTT assay was used to evaluate cell viability. DPPH scavenging activity was measured for antioxidation. Nitric oxide (NO) content was evaluated in lipopolysaccharide (LPS) treated RAW264.7 cells to indicate the performance on anti-inflammation. Results: In this study, six different compounds and their combinations were tested for melanogenesis. The results showed that the combination of glabridin, resveratrol, and ellagic acid (GRE) exhibited the highest efficiency, which was mainly manifested as inhibition of melanin production and tyrosinase activity, higher DPPH scavenging rate, and inhibition of nitric oxide (NO) production. Meanwhile, our results showed that GRE could significantly downregulate the expression of microphthalmia-associated transcription factor (MITF) related genes and proteins and could also inhibit the phosphorylation of cyclic AMP response element-binding protein (CREB), which was the upstream signal of MITF. Conclusion: The results suggest GRE exhibits high efficiency in inhibiting anti-melanogenesis, antioxidation, and anti-inflammation. Furthermore, GRE could downregulate the phosphorylation of the CREB and MITF signal pathway, which provides a theoretical basis for its application in pigmentation disorder disease and cosmetics.

Validating anti-inflammatory and cytotoxic properties of Fagonia cretica L. through metabolic, in vitro, and in silico profiling

BackgroundFagonia cretica L. (Family: Zygophyllaceae), is a wild shrub mostly found in Mediterranean districts and extensively used in folk medicine for a vast array of purposes such as antidiabetic and anticancer during the early stages. The goal of the current study was to validate the antioxidant, anti-inflammatory, and cytotoxic properties of Egyptian F. cretica using in vitro studies, metabolic profiling, and in silico approaches.MethodsThe plant was collected from the Egyptian desert and the alcoholic extract was prepared from its aerial parts, total phenolic and total flavonoid contents were evaluated spectrophotometrically. Antioxidant potential was assessed via 1,1 diphenyl-2-picrylhydrazyl (DPPH) scavenging activity. Anti-inflammatory activity was validated through in vitro COX-2, COX-1, and nitric oxide inhibition. Cytotoxicity was tested against liver (HepG2), breast (MCF-7), and intestinal (CACO2) carcinoma cell lines followed by assessment of its impact on the levels of apoptotic markers namely topoisomerase I and caspase 9 enzymes. Chemical profiling of the extract was performed using LC-HRMS technique. Saponin rich extract was prepared and tested for affecting topo I and caspase 9 enzymes. In silico studies were conducted on anti-inflammatory (COX-2 and COX-1) and cytotoxicity (topoisomerases I, IIα, and IIβ) targets using Autodock vina in PyRx platform.ResultsTotal phenolic and total flavonoid content of the extract were 2.4 ± 0.12 mg GAE/g and 0.18 ± 0.01 mg RE/g, respectively. In vitro results revealed antioxidant activity calculated as 1.4 ± 0.1 mg AEAC/g. In vitro anti-inflammatory assays unveiled inhibition of COX-2 and COX-1 enzymes with IC50 values of 13.02 ± 0.61 and 26.51 ± 0.83 µg/ml, respectively and nitric oxide with IC50 of 147.05 ± 9.61 µg/ml. Cytotoxicity on MCF-7, HepG2, and CACO2 cell line with IC50 values of 6.9 ± 0.53, 7.6 ± 0.42, and 9.2 ± 0.35 µg/ml, respectively, in addition to in vitro topoisomerase I inhibition (IC50 = 13.57 ± 0.71 µg/ml) and caspase 9 induction by 5.66 folds. Metabolic profiling using LC-HRMS technique resulted in dereplication of 21 compounds including triterpenoid saponins, flavonoids, diterpenoids, etc. Interestingly, saponin rich fraction and non-saponin fraction exhibited similar effects on topoisomerase I and caspase 9. In silico investigation unveiled high binding affinities of almost all the detected metabolites to the active sites of COX-2, COX-1, topo I, IIα, and IIβ enzymes.ConclusionCollectively, we can conclude that F. cretica is a new source of many phytochemicals, and a significant natural source as cytotoxic and anti-inflammatory agent.

Development of Herbal Ultrasound Gel Prototype from Zingiber cassumunar Roxb., Curcuma longa L. and Zingiber officinale Roscoe for Antioxidant and Anti-Inflammation

This research aimed to determine the total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity of phlai, turmeric, ginger and combined extract of phlai, turmeric, and ginger at recipe 1:2:2 (PTG). In addition, the formula of herbal ultrasound gel for antioxidant and anti-inflammatory properties was developed. The TPC, flavonoid, antioxidant and anti-inflammation were investigated by using the Folin-Ciocalteu method, flavonoid, DPPH, ABTS and nitric oxide (NO) scavenging assay. The PTG extract was tested for the phytochemicals by GC-MS technique. The microbial contamination standard of herbal ultrasound gel prototype was determined. The result show that the PTG extract has the TPC and flavonoid at 105.11 ± 16.93 mg GAE/g extract and 63.58 ± 2.29 mg QE/g extract, respectively. Moreover, the DPPH and ABTS assay show that PTG extract has the half maximal inhibitory concentration (IC50) at 0.82 ± 0.11 and 0.98 ± 1.56 mg/mL, respectively. In addition, PTG extract has a high percentage of NO inhibition at 53.38 ± 0.16. Furthermore, the GC-MS method found that PTG extract has 7 active ingredients. The highest amount of phytochemical is AR-turmerone. The safety assessment for external use found that the herbal ultrasound gel prototype product passed standard criteria for microbial contamination at the laboratory level. To conclude, the PTG extract has a synergistic effect and a high ability to act as an antioxidant and anti-inflammation. Moreover, the PTG herbal ultrasound gel has a microbial contamination safety to use. Therefore, the prototype herbal ultrasound gel product may be further developed into health products to treat pain and inflammation efficiently in clinical practice.

Antibacterial and Anti-Inflammatory Activity of Chitosan Film with Rhodomyrtus Tomentosa Leaf Extract Prepared Via 3D-Printing Method.

The combination of natural polymers and plant extract attracted much attention thanks to its valuable biological activities. This study presents the preparation and characterization of chitosan (CS) film containing Rhodomyrtus tomentosa leaf extract. In which, the Rhodomyrtus tomentosa leaf extract (SLE) with polyphenol-rich fractions was extracted by an ultrasonic-assisted extraction method. The prepared chitosan/polyphenol-rich fractions film (CSPPF) was characterized using infrared (IR) spectroscopy, mechanical testing, scanning electron microscopy (SEM), and water absorption analysis. Antibacterial and anti-inflammatory activities of the CSPPF were assessed using the agar well diffusion method and nitric oxide (NO) inhibition assays, respectively. Besides, the release of the extract from the CSPPF as well as the drug release kinetics was also tested. The Rhodomyrtus tomentosa leaf extract enhanced the antibacterial efficacy against Escherichia coli and enhanced the anti-inflammatory activity as well as the swelling ability of the CSPPF. The CSPPF containing 1 wt.% of SLE can inhibit 77.70 % NO with an IC50 value of 19.40 μg/mL.

Time-Dependent Relationship Between Endothelial Dysfunction and High Blood Pressure in Fructose Drinking Rats.

This study aims to investigate the duration required for endothelium dysfunction to develop in the fructose drinking-induced hypertension and examine the relative contributions of endothelium-dependent relaxing factors to changes in mesenteric arterial reactivity in male Wistar Albino rats. Metabolic parameters (water intake and food consumption) and hemodynamic parameters systolic blood pressure (SBP) and diastolic blood pressure (DBP)-were monitored in vivo. Vascular reactivity was examined in the isolated organ bath. Endothelium-dependent relaxation (EDR) to acetylcholine was observed in the absence and presence of pharmacological inhibitors of endothelial nitric oxide (NO) synthase, cyclooxygenase, and KCa2.3 channels. Contractile responses to phenylephrine and relaxation of sodium nitroprusside (SNP) were also determined. A significant increase in daily water intake and decrease in food consumption were typically observed in rats treated with 10% fructose for 4 weeks (p < 0.05). SBP and DBP increased significantly as early as 2 weeks of induction and continued to rise gradually throughout the induction period (p < 0.05). Fructose consumption significantly impaired EDR at week 3 and worsened at week 4 (p < 0.05). Impairment of the KCa3.1 channel-mediated component of endothelium-dependent hyperpolarisation (EDH)-type relaxation contributed to worsening EDR, whereas the contribution of NO-mediated relaxation was not apparent compared with the controls. The reduction in EDH-type relaxation in fructose-fed rats appears to be partially compensated by increased NO sensitivity in the smooth muscle region, as fructose induction increased SNP relaxation compared with the control. These data provide evidence of early endothelial dysfunction developing concurrently with increased blood pressure in 10% fructose-fed rats. Decreased KCa3.1-mediated part of EDH-type relaxation appears to contribute to the impairment of endothelium-dependent vasorelaxation over time in this model.

Endothelial-derived nitric oxide impacts vascular smooth muscle cell phenotypes under high wall shear stress condition

The Phenotypic states of vascular smooth muscle cells (SMCs) are essential to understanding vascular pathophysiology. SMCs in vessels generally express a specific set of contractile proteins, but decreased contractile protein expression, indicating a phenotypic shift, is a hallmark of vascular diseases. Recent studies have suggested the relation of abnormally high wall shear stress (WSS) of approximately 20 Pa with the aortic disease pathogenesis. However, due to the lack of appropriate experimental models to assess SMC phenotypic states, the details of the phenotypic shift under high WSS conditions remain unclear. In this study, we developed a coculture model where vascular endothelial cells (ECs) were cocultured with SMCs expressing calponin 1, a contractile protein involved in the phenotypic shift of SMCs. We investigated the effects of a pathologically high WSS condition on the phenotypic states of SMCs. Increased calponin 1 expression was found upon exposure to 20 Pa WSS compared with a physiological 2 Pa condition, whereas the expression of another contractile protein, α-smooth muscle actin (αSMA) remained unchanged. Furthermore, the inhibition of EC-derived nitric oxide (NO), which is associated with endothelial dysfunction in vascular diseases, resulted in a trend of decreasing αSMA and Calponin 1 expression under 20 Pa WSS conditions compared with 2 Pa. Our findings suggest that EC-derived NO under pathologically high WSS conditions may impact the expression of contractile proteins implicated in aortic pathophysiology.

Cardiospermum halicacabum Extract Inhibits the Production of Nitric Oxide and Reactive Oxygen Species in LPS-Stimulated RAW 264.7 Cells through Erk/p38 Signalling Pathways

The medicinal plant Cardiospermum halicacabum has been extensively studied for its potential in various therapeutic activities, including anti-inflammation, antioxidant, antibacterial, antifungal and antiparasitic effects. However, the specific molecular mechanisms responsible for its anti-inflammatory properties remain largely unknown. In order to shed light on this aspect, the current investigation was aimed at elucidating the underlying mechanisms through which C. halicacabum exerts its anti-inflammatory response. The different concentrations of C. halicacabum extract were tested for cytotoxicity on RAW264.6 murine macrophage cells by MTT assay. The inhibition of nitric oxide (NO) in LPS-induced cell pre-treated with the extract was evaluated by Griess assay using Griess reagent. Flow cytometry and fluorescent microscopy were used to examine the reactive oxygen species (ROS) in the treated cells. The expression of iNOS and the phosphorylation status of Erk1/2, p38 and NF-kB were determined by western blotting. The C. halicacabum extract at concentrations up to 125 µg/mL did not cause the cytotoxicity to the RAW264.6 murine macrophage cells. Incubation of LPS-induced cells with the extract significantly minimized the production of NO as compared with the LPS-stimulated cells. Further, the extract significantly reduced ROS generation in LPS-induced cells. In the present of C. halicacabum, LPS-treated cells had the reduced NOS expression as well as had low phosphorylation on Erk1/2, p38 and NF-kB. C. halicacabum attenuates NO and ROS production under the regulation of Erk1/2 and p38 pathway as well as NF-kB. HIGHLIGHTS The underlying mechanisms of halicacabum on its anti-inflammatory response were investigated. halicacabum extract significantly reduced ROS and NO generation in LPS-induced cells. halicacabum attenuates NO and ROS production under the regulation of Erk1/2 and p38 pathway as well as NF-kB. GRAPHICAL ABSTRACT

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. In addition to 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: did 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.

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.

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

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.

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.