Griess Reaction Research Articles

Viologen-Radical-Driven Hydrogen Evolution from Water Catalyzed by Co-NHC Catalysts: Radical Scavenging by Nitrate and Volmer-Heyrovsky-like CPET Pathway.

The factors controlling the catalytic activity in photochemical hydrogen evolution reaction (HER) are studied in detail for two macrocyclic cobalt compounds bearing two N-heterocyclic carbenes and two pyridyl donors (Co-NHC1 and Co-NHC2, where Co-NHC2 has a methoxy substituent on each pyridyl ligand). The present study adopts an aqueous photosystem consisting of EDTA, [Ru(bpy)3]2+ (bpy = 2,2'-bipyridine), and MV2+ (MV2+ = methylviologen) at pH = 5. Both catalysts are shown to promote HER in a similar efficiency (TON = 12-13 in 6 h), revealing a minor contribution of the electron-donating methoxy substituents. The catalyst degradation is shown to proceed during the photocatalysis, leading to afford [Co(edta)]- (EDTA = H4edta) as a dead-end species. The lack of any heterogeneous species was evidenced by DLS (dynamic light scattering). It was also found that nitrate involved as a counteranion in the photocatalysis components substantially inhibits the photocatalytic HER, giving rise to a large diminishment in TON from 12.7 to 7.2. The Griess test was used to confirm that NO3- serves as a scavenger deactivating the reduced form of MV2+ (i.e., MV+·). The detailed spectroscopic study reveals that the radical dimer (MV+·)2 plays a key role in promoting the one-step two-electron process: (MV+·)2 + NO3- + 2H+ → 2MV2+ + NO2- + H2O. Experimental and DFT results also reveal that a unique double CPET (concerted proton-electron transfer) pathway is taken to evolve H2 by the Co-NHC catalysts with substantially minimized reorganization energies: Co(II)-NHC Co(III)(H)-NHC Co(II)-NHC + H2. This pathway can be viewed as related to the so-called Volmer-Heyrovsky mechanism adopted by some metals and is quite unique to the Co-NHC catalysts.

Immunomodulatory Effects of Pterocarpanquinone LQB-118 in Murine Peritoneal Macrophages

ABSTRACT Background Phagocytosis is an important function of macrophages. However, when it’s dysregulated, it could compromise homeostasis. Thus, this study aimed to assess the inhibitory activity of pterocarpanquinone LQB 118 on murine macrophage phagocytosis. Methods We used peritoneal macrophages isolated from mice to evaluate the impact of LQB 118 (5 μM) on the modulation of phagocytic activity and possible action mechanism related: IL-12 (by ELISA), NO (by Griess reaction),ROS production (by flow cytometry), and intracellular signaling proteins (iNOS, P-Akt, P-mTOR, NF-κB, and P-NF-κB) (by flow cytometry).The macrophages were stimulated with zymosan to assess both phagocytic activity and flow cytometry assays. Results Treatment with LQB 118 resulted in a reduction in the phagocytosis of zymosan particles by macrophages. This effect could potentially be attributed to LQB’s inhibition of IL-12 production and mTOR/NF-κB signaling. Furthermore, LQB 118 decreased the levels of ROS and NO without interfering with iNOS and Akt activation. Conclusion These findings show the anti-phagocytic activity of LQB 118 on macrophage, highlighting the potential of this compound as a candidate for modulating macrophage-driven inflammation.

Association of NOS2A Gene Polymorphisms with Susceptibility to Tuberculosis in Manipuri Population of Northeast India.

Single nucleotide polymorphisms (SNPs) have been reported to influence the activity of specific genes involved with the innate immune response to Mycobacterium; hence, they are crucial in tuberculosis (TB) susceptibility studies. The study aimed to investigate the polymorphism in the NOS2A (Nitric oxide synthase 2A)geneand its association with susceptibility to TB in the Manipuri population of northeast India. This case-control study includes 495 subjects- 220TB patients and 275 control individuals. TaqMan allelic discrimination assay was used to study the gene polymorphism, and Griess's test was employed to determine the serum nitric oxide (NO) levels. Serum NO levels were analysed to correlate with the functional changes associated with the polymorphisms. Two SNPs of the gene,NOS2A(rs8078340 and rs2274894), were studied. For the SNP-rs8078340, a significant difference in the genotypic and allelic frequencies was observed between the cases and control groups (p = 0.001; AA genotype OR = 30.288, 95% CI: 1.703-538.44 and A allele OR = 2.937, 95%CI: 1.762-4.896). However, for the SNP-rs2274894, only the T allele (with OR = 1.464; 95%CI: 1.080-1.983, p = 0.014) was associated with susceptibility to TB. Serum levels of NO were significantly different between the cases and control groups (p < 0.05). Significant associations of both homozygous AA genotype and allele A of the NOS2A (rs8078340) and minor allele T of NOS2A (rs2274894) were observed with susceptibility to TB. Patients with the AA genotype of NOS2A show a higher NO level, suggesting its role in greater expression of the NOS2A gene.

Deficiency of Klotho protein as a cause of linked angina in young patients with thoracalgic phenotype of gastroesophageal reflux disease

Gastroesophageal reflux disease (GERD) is one of the most common diagnoses in therapeutic practice. Different manifestations of GERD represent different phenotypes of the disease with unique predisposing cofactors and individual pathophysiology. In the continuum of the proposed paradigm, chest pain syndrome in patients with gastroesophageal reflux stands out as a separate phenotype of GERD. Traditionally it was believed that the chest pain in young patients with GERD was not angina. Objective — to study the levels of Klotho protein and nitric oxide in the blood serum of patients with GERD and chest pain (thoracalgic phenotype of GERD). Materials and methods. 64 patients with GERD were prospectively examined, including 27 patients with the thoracalgic phenotype of GERD. The Klotho protein was determined in blood serum by the enzyme immunoassay, the metabolism of NO in the blood plasma was assessed by the Griess test. All patients underwent video esophagogastroscopy, transthoracic echocardiography and 24­Hour Holter electrocardiographic monitoring. Descriptive and nonparametric statistics methods were used in the study. Results. An evaluation of the chest pain syndrome in young patients with GERD revealed a high incidence of linked angina — 25.9%. A decrease in the Klotho protein and NO levels in the blood of patients with GERD and linked angina was observed compared to patients with uncomplicated GERD with pseudo­­coronary pain. The obtained result showed a direct relationship between the concentrations of Klotho protein and NO. Low levels of these parameters in patients with GERD and linked angina reflect endothelial dysfunction and lead to coronary artery spasm mediated by reflux. Conclusions. Linked angina occurs in a quarter of young patients with the thoracalgic phenotype of GERD. The basis of this phenomenon is a decrease in NO synthesis due to endothelial dysfunction, which develops against the background of the Klotho protein deficiency.

Investigation of ROS-Driven Cytotoxic Mechanisms in WO3:Ag Heterostructures Supported on Carbon Against Bladder Cancer

Bladder cancer presents a significant health challenge due to its high malignancy and rising incidence rates. Silver-based materials are well-known for their cytotoxic effects on various cell types. This study not only aimed to synthesize and characterize carbon-supported WO3:Ag heterostructures but also to evaluate their biological and physicochemical properties. The material was synthesized through the synergistic thermal decomposition of α-Ag2WO4 dispersed in chitosan, followed by WO3:Ag heterostructure formation on a carbon support, yielding samples with varying α-Ag2WO4 concentrations (SC, SC1, SC2, and SC4, for 0, 10, 20 and 40% of α-Ag2WO4 for chitosan). Characterization confirmed the successful formation of carbon-supported heterostructures with controlled ionic release and enhanced ROS generation. In vitro assays were conducted to assess the viability of non-tumor (3T3 fibroblasts) and tumor (bladder carcinoma MB49) cells using MTT salt and neutral red dye. Additional analyses included autophagy detection by correlating data from viability assays, nitric oxide and ROS quantification using the Griess reaction and fluorescent probes, and Caspase-3 activity measured with a fluorescent antibody. The results indicated that SC1 and SC2 samples were more effective against both cell types, with SC2 showing heightened effectiveness against the tumor lineage by inducing greater oxidative stress in MB49 cells compared to 3T3 fibroblasts. Additionally, the materials exhibited low ionic release (&lt;0.01%), reducing potential adverse effects. Mechanistic analysis showed that the carbon support and synergistic interactions between WO₃ and Ag modulated ⦁OH radical production, even without light, enhancing the material's cytotoxic efficiency. These findings highlight the therapeutic potential of WO₃:Ag heterostructures as a safe and effective approach for treating aggressive cancers like bladder carcinoma, emphasizing the importance of further development in advanced biofunctional materials. This study also highlights the therapeutic potential of carbon-supported WO3:Ag heterostructures in bladder cancer treatment and underscores the importance of continued research in the development of novel anticancer strategies.

Cytotoxic, Antioxidant, and Anti-inflammatory Effects of Crataegus meyeri

Background: Crataegus meyeri A. Pojark is one of Iran›s most well-known medicinal plants. This study aimed to assess the cytotoxic, antioxidant, and anti-inflammatory properties of the methanolic extract of Crataegus meyeri (MCM) leaves. Methods: MCM was prepared from dried leaves. First, the cytotoxic impact of MCM was determined via the MTT test. Then, the anti-inflammatory activity of MCM on the lipopolysaccharide (LPS)-exposed J774A.1 cells was examined by measuring NO production and cyclooxygenase-2 (COX-2) gene expression using the Griess test and quantitative real-time polymerase chain reaction (qRT-PCR), respectively. MCM antioxidant activity was assessed using the FRAP and DPPH tests. Results: MCM was shown to be non-cytotoxic against J774A.1 cells at≤125 and≤62 μg/mL concentrations after 24 and 48 hours, respectively. It also significantly attenuated NO generation in LPS-treated J774A.1 cells at 62 and 125 μg/mL after 24 hours and 62 μg/mL after 48 hours. The extract significantly reduced COX-2 gene expression at concentrations of 62 and 125 μg/mL after 24 hours and at concentrations of 15, 31, and 62 μg/mL after 48 hours. Moreover, the results indicated that MCM at all concentrations exhibited lower antioxidant activity than vitamin C, but the difference was insignificant. Furthermore, MCM could reduce the viability of most of the studied cancer cell lines at concentrations of 250 and 500 μg/mL. Conclusion: The findings of the present study suggest that MCM may show therapeutic potential for the treatment of inflammatory diseases, a finding that requires clinical trials.

Integrating Chromophoric and Instrumental Methods (SEM-EDS and FTIR) for Accurate Shooting Distance Estimation: An Inter-laboratory Study in Forensic Chemistry

Background: Accurate shooting distance estimation is critical in forensic investigations. Traditional color tests like the Modified Griess Test (MGT) and Sodium Rhodizonate Test (SRT) are commonly used but lack sensitivity and objectivity, necessitating improvements in reproducibility and validation. Methods: A 7.62x39 mm cartridge was fired using an assault rifle (Sarsılmaz® SAR 308) at fabric samples from distances ranging from 0 to 100 cm. MGT and SRT were employed to estimate shooting distances, and the presence of inorganic and organic gunshot residues was confirmed using Scanning Electron Microscopy with Energy Dispersive X-ray Spectroscopy (SEMEDS) and Fourier Transform Infrared Spectroscopy with Attenuated Total Reflection (FTIRATR). Inter-laboratory reproducibility was assessed through Relative Standard Deviation (RSD), paired t-tests, and Pearson Correlation Coefficient (PCC). Results: MGT and SRT showed high reproducibility at short distances (RSD &lt;11% up to 30 cm), but sensitivity decreased at longer distances, with an RSD of 28.6% for SRT at 100 cm. Paired t-tests revealed statistically significant consistency between laboratories (p &lt; 0.05), and a strong negative correlation (PCC = -0.72) was found between shooting distance and residue area. method: This research supports the use of SEM-EDS and FTIR to improve the accuracy of these color tests, providing improved forensic analysis of 7.62x39 mm cartridge firearms incidents. Conclusion: Integrating SEM-EDS and FTIR-ATR with MGT and SRT enhances the sensitivity, objectivity, and reproducibility of shooting distance estimations. This multi-method approach provides more reliable forensic evidence for criminal investigations. result: The inter-laboratory accuracy of these methods was evaluated, and SRT and MGT showed relative standard deviations of 9.48% and 10.30%, respectively.

Agaricus bisporus mannose-binding protein stimulates the innate immune cells

Purpose: A lectin-like protein from the mushroom Agaricus bisporus has been shown to slightly increase the proliferation of RAW 264.7 cells. Following its identification as a mannose-binding lectin, henceforth called A. bisporus mannose-binding protein (Abmb), the protein is hypothesized to stimulate the innate immune cells response. The present work was aimed to substantiate that hypothesis. Furthermore, this study complements Abmb exploration as a potential agent for anti-breast cancer, which its treatment is hampered with compromised immunity of patient receiving chemotherapy. Method: Abmb’s effect on the phagocytic activity of the macrophage was measured with FACS. Nitric oxide production was checked using Griess test while expression of the cytokines in the RAW 264.7 cells was analysed at gene and protein level using PCR and FACS, respectively. Abmb’s effect on the expression of surface markers of the human immune cells in the peripheral blood mononuclear cells was checked with specific antibodies for targeted cluster differentiation and analysed using FACS. Result: Abmb increased the phagocytic activity of the macrophage and NO production. Abmb increased the expression of cytokines i.e. tumour necrosis factor (TNF)-α, interleukin (IL)-6, and IL-10. With the peripheral blood mononuclear cells, Abmb activated dendritic and Natural Killer cells, but not the B- or T-cells. Conclusion: Abmb increased the activity of the macrophage cells and activated the immune cells that are related to the innate immune system, particularly the cellular immunity.

Exploring the unique antioxidative and anti-inflammatory properties of aging black lemon (Citrus limon L. Brum. F.)

The aging black lemon is a dried citrus fruit, generally made from lemon, which is lightly thermal processed until it takes on the appearance of blackish-brown leather. This study aimed to determine the phenolic and flavonoid components in aqueous black lemon extract (ABLE). FRAP was used to determine the antioxidant capacity of ABLE. The effects of ABLE on LPS-induced NO production in macrophages (RAW264.7) were investigated by measuring the amount of nitrite released into the culture medium using the Griess reaction. For the anti-inflammatory effects, pro-inflammatory mediators, including iNOS, TNF-α, and NF-κB, were examined by Western blotting. In addition, UPLC/MS/MS data of ABLE presented higher concentrations of gallic acid, protocatechuic acid, vanillic acid, p-coumaric acid, ferulic acid, luteolin, naringenin, hesperetin, and diosmetin than fresh lemon. Based on the results, we consider ABLE to have outstanding anti-inflammation properties by polyphenols and flavonoids and is a great functional food for inflammation-related diseases.

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

Ganoderma lucidum polysaccharide alleviates cognitive dysfunction by inhibiting neuroinflammation via NLRP3/NF-κB signaling pathway

Ethnopharmacological relevanceGanoderma lucidum (G. lucidum), a traditional Chinese medicinal herb, is commonly recommended for its potential to promote mental relaxation and alleviate memory impairment. Recently, there have been reports suggesting that it exhibits anti-neuroinflammatory activity through the gut-brain axis. Cognitive dysfunction is among the most prevalent neurodegenerative diseases. Aim of the studyThis study aimed to investigate the efficacy of polysaccharides extracted from G. lucidum in alleviating cognitive dysfunction. Methods and materialsA polysaccharide was extracted through the process of alkali extraction followed by alcohol precipitation. Comprehensive analysis was conducted to characterize the total sugar content, amino acid composition, and sugar chain structure. The levels of inflammatory related factors were assessed using griess reagent, qPCR and western blotting assay in vitro. The efficacy of alleviating cognitive dysfunction was evaluated through a series of behavioral studies in mice model induced by the high-fat high-sugar diet combined with chronic unpredictable mild stress (HFFD/CUMS) in vivo. The mechanism was investigated by 16S rRNA sequence, immunohistochemistry, flow cytometry and short-chain fatty acid detection. ResultsGanoderma lucidum polysaccharide (GLP) is a polysaccharide identified as β-glucan. Bioactivity experiments have demonstrated that GLP possesses the potential to ameliorate cognitive dysfunction. The mechanism study revealed that GLP can modulate the composition of gut microbiota and suppress the activation of inflammasomes via NLRP3/NF-κB signaling pathway, thereby attenuating neuroinflammatory. Furthermore, GLP may enhance the peripheral immunity response of the body, leading to a comprehensive regulatory effect. ConclusionA polysaccharide alleviates cognitive dysfunction via inhibiting neuroinflammation.

In vitro and in vivo evaluation of anti-inflammatory activities of ethanol extract from Lom-Am-Ma-Pruek remedy for pain relief

Background and purpose: Lom-Am-Ma-Pruek (LAMP) remedy has been used in Thai traditional medicine to relieve pain associated with the inflammatory process. The anti-inflammatory activity and bioactivity of LAMP in an animal model have not been previously investigated. We evaluated the in-vitro and in-vivo anti-inflammatory activity of LAMP ethanol extract. Experimental approach: The anti-inflammatory activity of LAMP and its plant ingredients were investigated on lipopolysaccharide-stimulated NO, PGE2, and TNF-α release from RAW264.7 cells. Furthermore, the stability of LAMP under biological and chemical accelerated conditions was evaluated using the Griess reaction assay and HPLC. Lastly, rat models with ethyl phenylpropionate (EPP)-induced ear edema and carrageenan-induced paw edema were utilized to assess anti-inflammatory activity. Findings/Results: LAMP possessed potent inhibitory effects on NO, PGE2, and TNF-α production with IC50 values of 24.90 ± 0.86, 4.77 ± 0.03, and 35.01 ± 2.61 µg/mL, respectively. In addition, LAMP extract demonstrated stable biological activity, anti-inflammatory effects, and phytochemical content stability under stress conditions. Additionally, 0.5%, 1%, and 2% w/v LAMP significantly inhibited EPP-induced rat ear edema over time equivalent to 5% w/v phenylbutazone. LAMP at 180, 375, and 750 mg/kg also considerably reduced carrageenan-induced rat paw edema 2 h after carrageenan administration compared to phenylbutazone at 250 mg/kg. Conclusion and implications: LAMP has anti-inflammatory activity by inhibiting PGE2 formation. These findings are consistent with the efficacy and traditional use of the LAMP remedy in treating inflammatory diseases.

Melissa officinalis Regulates Lipopolysaccharide-Induced BV2 Microglial Activation via MAPK and Nrf2 Signaling.

Neuroinflammation and microglial activation play critical roles in neurodegenerative diseases such as Alzheimer's and Parkinson's disease. Modulating microglial activation may help prevent the progression of these disorders. This study aimed to investigate the effects and mechanisms of Melissa officinalis ethanol extract on lipopolysaccharide (LPS)-induced microglial activation in BV2 cells. Cell viability and nitric oxide (NO) production were assessed using MTT assay and Griess reagent, while inflammatory cytokine levels were measured by qPCR. Key inflammatory pathways, including MAPK, TLR4, and antioxidant biomarkers, were analyzed through western blot and immunofluorescence. Rosmarinic acid content in M. officinalis was determined using high-performance liquid chromatography (HPLC). The results demonstrated that M. officinalis ethanol extract significantly inhibited LPS-induced NO production and reduced inflammatory cytokine expression. Additionally, it downregulated inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), TLR4, NF-κB, and MAPK signaling pathways (p38, JNK, ERK), while increasing the expression of antioxidant markers, including Nrf2, HO-1, catalase, and SOD2. In conclusion, M. officinalis ethanol extract exerts neuroprotective effects by modulating inflammation and enhancing antioxidant defenses, suggesting its potential in the prevention and treatment of inflammation-related neurodegenerative diseases.

Acute Chikungunya Infection Induces Vascular Dysfunction by Directly Disrupting Redox Signaling in Endothelial Cells.

Chikungunya virus (CHIKV) infection is characterized by febrile illness, severe joint pain, myalgia, and cardiovascular complications. Given that CHIKV stimulates reactive oxygen species (ROS) and pro- and anti-inflammatory cytokines, events that disrupt vascular homeostasis, we hypothesized that CHIKV induces arterial dysfunction by directly impacting redox-related mechanisms in vascular cells. Wild-type (WT) and iNOS knockout (iNOS-/-) mice were administered either CHIKV (1.0 × 106 PFU/µL) or Mock vehicle via the intracaudal route. In vivo, CHIKV infection induced vascular dysfunction (assessed by a wire myograph), decreased systolic blood pressure (tail-cuff plethysmography), increased IL-6 and IFN-γ, but not TNF-α levels (determined by ELISA), and increased protein content by Western blot. Marked contractile hyporesponsiveness to phenylephrine was observed 48 h post-infection, which was restored by endothelium removal. L-NAME, 1400W, Tiron, and iNOS gene deletion prevented phenylephrine hyporesponsiveness. CHIKV infection increased vascular nitrite concentration (Griess reaction) and superoxide anion (O2•-) generation (lucigenin chemiluminescence), and decreased hydrogen peroxide (H2O2, by Amplex Red) levels 48 h post-infection, alongside increased TBARS levels. In vitro, CHIKV infected endothelial cells (EA.hy926) and upregulated ICAM-1 and iNOS protein expression (determined by Western blot). These data support the conclusion that CHIKV-induced alterations in vascular ROS/NF-kB/iNOS/NO signaling potentially contribute to cardiovascular events associated with Chikungunya infection.

Stachys byzantina K. Koch (Lamiaceae) as a potential ingredient for delaying skin ageing and treating hyperpigmentation disorders in pharmaceutical products.

This study aimed to investigate whether the plant species Stachys byzantina produces bioactives with the potential to delay the skin ageing process and treat hyperpigmentation conditions. The antioxidant action was assessed by 2,2-diphenyl-1-picrylhydrazylradical scavenging, Griess reaction, oxygen radical absorption capacity, and β-carotene bleaching assays. Inhibitory activities for tyrosinase, hyaluronidase, and elastase enzymes were tested. The antiglycation activity, the sun protection factor (SPF), and the toxicity to skin cells by MTT(3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide)assay were also evaluated. The ethanolic extract of S. byzantina aerial parts and all fractions obtained by solvent partition inhibited the tyrosinase enzyme at different levels. The dichloromethane fraction (DF) demonstrated the highest inhibition (IC50 = 63.5±10.9 µg/ml). DF also inhibited the hyaluronidase enzyme with IC50 = 369±11.64 μg/ml and elastase by 40% at 500 μg/ml. This fraction showed prominent antioxidant and antiglycation activities, high SPF, and no cytotoxicity at concentrations lower than 50 μg/ml. The phenolic and flavonoid contents were 116.30±6.7 (mgTAE/g) and 66.38±13.5 (mgQE/g), respectively. Chlorogenic acid (23.54±2.46mg/g) and verbascoside (203.97±19.8mg/g) were identified and quantified. Stachys byzantina is a potential source of cosmetic and therapeutic ingredients to reduce hyperpigmentation and the impacts caused by free radicals, advanced glycation end products, and sun radiation in skin ageing.

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.

Lignans of Schisandra chinensis (Turcz.) Baill inhibits Parkinson's disease progression through mediated neuroinflammation-TRPV1 expression in microglia

BackgroundSchisandra chinensis (Turcz.) Baill (S. chinensis), a member of the Magnoliaceae family, is renowned for its distinctive medicinal attributes and is commonly employed in the treatment of disorders affecting the CNS. PurposeThe potential therapeutic effects of a lignan-enriched extract derived from Schisandra chinensis (Turcz.) Baill (LSC) on PD is assessed, which focuses on its mechanisms of action in addressing neuroinflammation. MethodsThe LSC has been obtained by purifying the ethyl alcohol extract of S. chinensis. The Orbitrap-MS method has been employed to analyze the chemical composition of the LSC. In LPS-induced BV2 cells, LSC-induced changes in M1/M2 type inflammatory cytokines have been examined using the Griess reaction, Elisa, JC-1, flow cytometry, IF, and WB methods. A model of PD has been established by treatment of MPTP in C57BL/6 mice. The effect of LSC on behavioral changes, inflammatory factor levels, expression of TH and IBA-1, and production of autophagy in the midbrain has been investigated by TEM, immunohistochemistry, Elisa, and WB. ResultsLSC has relieved sports injuries and pathological damage, and targeted the TRPV1-AMPK-NLRP3 signaling pathway, which affected neuroinflammation and autophagy in vivo. Furthermore, in vitro investigations demonstrated that LSC has activated M1/M2 transformation, its related inflammatory factors, and protein expressions of the NLRP3-Caspase1 signaling pathway in LPS-BV2 cells. The research notably demonstrated that the LSC promoted autophagy and suppressed inflammation through targeting TRPV1. ConclusionIn the investigation, LSC focused on TRPV1 and controlled neuroinflammation-autophagy by regulating AMPK-NLRP3, which has been proven for the first time. The study has presented molecular data supporting the use of LSC in treating PD and offers references for developing drugs. Remarkably, LSC has the potential to be utilized as a therapeutic or health medication that could significantly decrease PD.

&lt;i&gt;Oroxylum indicum&lt;/i&gt; Kurz (L) leaf extract exerted antioxidant and anti-inflammatory effects on LPS-stimulated BV2 microglial cells

Purpose: To investigate the antioxidant and anti-inflammatory effects of Oroxylum indicum (L.) Kurz leaf extract on lipopolysaccharide (LPS)-activated BV2 microglia. Methods: BV2 cells were treated with LPS for 24 h in the presence or absence of Oroxylum indicum (O. indicum) leaf extract. The levels of reactive oxygen species (ROS), nitric oxide (NO), and interleukin 6 (IL-6) were quantified with CM-H2DCFDA, Griess reagent assay, and enzyme-linked immunosorbent assay (ELISA) test, respectively. The antioxidant activity of O. indicum leaf extract was assessed in vitro using DPPH, ABTS and FRAP assays. Results: Treatment with LPS resulted in significant increases in the levels of ROS, NO, and IL-6, when compared to untreated cells. However, co-treatment with O. indicum leaf extract significantly suppressed the production of these inflammatory markers (p &lt; 0.05). In addition, the leaf extract of O. indicum exhibited antioxidant activity in the DPPH, ABTS, and FRAP assays. The contents of flavonoids and phenolics in O. indicum leaf extract were 13.25 ± 0.163 mg QE/g of dried extract and 82.58 ± 1.772 mg QE/g of dried extract, respectively (p &lt; 0.05). Conclusion: The leaf extract of O. indicum exhibits anti-inflammatory and antioxidant properties in LPS-activated BV2 cells. This finding indicates that the leaf extract of O. indicum has promising potentials as a new source of herbal medication with anti- inflammatory properties. However, there is a need for more studies to unravel the fundamental mechanisms of their actions.

Proteomics fingerprinting reveals importance of iron and oxidative stress in Streptomyces scabies-Solanum tuberosum interactions.

The Gram-positive actinobacterium Streptomyces scabies is the major causal agent of potato common scab. The main pathogenicity factor is thaxtomin A, a phytotoxin that causes atypical cell death, although other secondary metabolites have been described to play a role in S. scabies virulence. Despite this, many aspects of the interaction between S. scabies and its primary host Solanum tuberosum L. remain to be elucidated. Intracellular proteins of S. scabies EF-35 grown in the presence of in vitro produced tubers (microtubers) of the Russet Burbank and Yukon Gold potato cultivars were extracted and analysed by electrospray mass spectrometry (ES MS/MS). Based on the results of proteomic analysis, iron quantification by ICP-MS and nitrite quantification using Griess reagent in growth media as well as RT-qPCR analysis of the siderophore pyochelin gene expression were performed in the presence and absence of microtubers. Hydrogen peroxide accumulation was also determined in the nutrient medium used for co-cultivation of bacteria and potato microtubers. Potato microtubers caused an increase in the content of bacterial proteins involved in stress and defense, secondary metabolism, and cell differentiation, as well as secreted proteins. Co-cultivation with potato microtubers induced the accumulation of S. scabies proteins implicated in siderophore pyochelin biosynthesis, nitrite production and oxidative stress perception and response. The increase in the abundance of proteins related to pyochelin biosynthesis was consistent with a significant decrease in the iron content in the culture medium, as well as with induction of expression of pyochelin biosynthesis genes. Elevated nitrite/sulfite reductase protein levels were associated with increased nitrite excretion by S. scabies cells in the presence of host microtubers. The increase in the levels of proteins associated with signaling and oxidative stress response could have been caused by the accumulation of ROS, in particular hydrogen peroxide, detected in the studied system. These findings show that interactions of S. scabies with living potato microtubers induce the production of secondary metabolites, defense responses, and protection from oxidative stress. This study suggests the importance of iron during host - S. scabies interactions, resulting in competition between pathogen and its host.

Spermidine attenuates chondrocyte inflammation and cellular pyroptosis through the AhR/NF-κB axis and the NLRP3/caspase-1/GSDMD pathway.

Osteoarthritis (OA) is a prevalent chronic degenerative disease, marked by a complex interplay of mechanical stress, inflammation, and metabolic imbalances. Recent studies have highlighted the potential of spermidine (SPD), a naturally occurring polyamine known for its anti-inflammatory and antioxidant properties, as a promising therapeutic agent for OA. This study delves into the therapeutic efficacy and mechanistic pathways of SPD in mitigating OA symptoms. Forty Sprague-Dawley rats were randomly assigned to four groups, including the CG (sham operation), model (anterior cruciate ligament transection [ACLT], and treatment (ACLT + two different doses of SPD) groups. In vivo, correlations between OA severity and different interventions were assessed by ELISA, X-rays, CT imaging, histological staining, and immunohistochemistry. In vitro, IL-1β was used to trigger chondrocyte inflammation, and SPD's cytotoxicity was assessed in primary rat chondrocytes. Next, inflammatory markers, extracellular matrix (ECM) proteins, and pathway marker proteins were detected in chondrocytes administered IL-1β alone, SPD, or aryl hydrocarbon receptor (AhR) silencing, by qRT-PCR, Griess reaction, ELISA, Western blot, and immunofluorescence. Morphological alterations and pyroptosis in chondrocytes were examined by transmission electron microscopy (TEM) and flow cytometry. Our research reveals that SPD exerts significant anti-inflammatory and antipyroptotic effects on IL-1β-treated chondrocytes and in anterior cruciate ligament transection (ACLT) rat models of OA, primarily through interaction with the Aryl hydrocarbon receptor (AhR). Specifically, SPD's binding to AhR plays a crucial role in modulating the inflammatory response and cellular pyroptosis by inhibiting both the AhR/NF-κB and NLRP3/caspase-1/GSDMD signaling pathways. Furthermore, the knockdown of AhR was found to negate the beneficial effects of SPD, underscoring the centrality of the AhR pathway in SPD's action mechanism. Additionally, SPD was observed to promote the preservation of cartilage integrity and suppress ECM degradation, further supporting its potential as an effective intervention for OA. Collectively, our findings propose SPD as a novel therapeutic approach for OA treatment, targeting the AhR pathway to counteract the disease's progression and highlighting the need for further clinical evaluation to fully establish its therapeutic utility.