Activity In RAW Research Articles

Metal-Free CO Prodrugs Activated by Molecular Oxygen Protect against Doxorubicin-Induced Cardiomyopathy in Mice.

Carbon monoxide has been extensively studied for its various therapeutic activities in cell cultures and animal models. Great efforts have been made to develop noninhalational approaches for easy and controlled CO delivery. Herein, we introduce a novel metal-free CO prodrug approach that releases CO under near-physiological conditions. CO from the quinone-derived CO prodrugs is initiated by general acid/base-catalyzed tautomerization followed by oxidation by molecular oxygen to form the key norbornadienone intermediate, leading to cheletropic CO release only in an aerobic environment. Representative CO prodrug analog QCO-105 showed marked anti-inflammatory effects and HO-1 induction activity in RAW264.7 macrophages. In a mouse model of doxorubicin-induced cardiomyopathy, we show for the first time that the CO prodrug QCO-105 prevented cardiomyocyte injury, consistent with the known organ-protective effects of HO-1 and CO. Overall, such a new CO prodrug design serves as the starting point for developing CO-based therapy in attenuating the cardiotoxicity of doxorubicin.

A water-soluble mycelium polysaccharide from Monascus pilosus: Extraction, structural characterization, immunomodulatory effect and yield enhanced by overexpression of UGE gene

Mycelium polysaccharide (MPP) from Monascus pilosus with the compositions of glucose, galactose, mannose, glucosamine hydrochloride, rhamnose and arabinose, was obtained using alkaline extracting, and subsequently three purified components (MPP-0, MPP-0.1 and MPP-0.3) were separated. The purity and extraction volume of the MPP-0.1 fraction surpassed those of the other two groups, thus warranting its selection for subsequent experimental investigations. The sample MPP-0.1, with an average molecular weight of 3.7776 × 104 Da, exhibited exceptional thermal stability up to 170 °C. The main glycosidic linkage pattern of MPP-0.1 was structured as→[4)-α-D-Glcp-(1]6 → 4)-α-D-Glcp-(1 → [2)-α-D-Manp-(1]5 → 2)-α-D-Manp-(1 → 5)-β-D-Galf-(1 → 3)-β-D-Galf (1 → 3)-β-D-Galf-(1 → 3)-β-D-Galf-(1→, and branched Glcp, Manp, Galf fragments were connected with the main chain through →4, 6)-α-D-Glcp-(1→, →2, 6)-α-D-Manp-(1 → and →3, 6)-β-D-Galf-(1→. Besides, the up-regulated levels of Nitric oxide (NO), Tumor necrosis factor-α (TNF-α), Interleukin-6 (IL-6), Interleukin-1β (IL-1β) and other pro-inflammatory cytokines along with increased phagocytic activity revealed that MPP-0.1 has significant immunomodulatory effect, and can significantly enhance the proliferation and activation of RAW264.7 cells. Finally, the gene UGE (UDP-glucose 4-epimerase) was overexpressed in M. pilosus to increase the MPP production. Results showed that the biomass of the recombinant strain exhibited a remarkable increase of approximately 62.56 ± 1.50 % compared to that of the parental strain, and the extraction yield of MPP increased significantly by 83.19 ± 4.56 %.

Immunostimulatory activity of the aqueous extract from the leaves of Sambucus racemosa subsp. pendula through TLR4‑dependent JNK activation in RAW264.7 cells.

Sambucus racemosa subsp. pendula (SRP) is an endemic plant of Korea, exclusively found on Ulleungdo Island. SRP is widely used as both a traditional medicine and food source. However, there is a lack of research on the pharmacological activities of SRP. Therefore, the present study aimed to explore the potential use of SRP leaves (SRPL) as a natural immunostimulant by analyzing its macrophage activation properties and the underlying mechanisms of action. Among the various extraction conditions, SRPL (AE20-SRPL) extracted with 100% distilled water at 20˚C induced the highest nitric oxide (NO) production in RAW264.7 cells. Thus, the further studies were performed using AE20-SRPL. AE20-SRPL increased the production of immunostimulatory factors such as NO, prostaglandin E2, inducible nitric oxide synthase, cyclooxygenase-2, IL-1β and TNF-α and phagocytosis in a dose-dependent manner in RAW264.7 cells without exhibiting cytotoxicity. Among Toll-like receptor (TLR)2 and TLR4, inhibition of TLR4 significantly reduced AE20-SRPL-mediated increases in the production of immunostimulatory factors and phagocytosis in RAW264.7 cells. Furthermore, in RAW264.7 cells, inhibition of JNK, one of the components of MAPK signaling along with ERK1/2 and p38, attenuated the AE20-SRPL-mediated increases in the production of immunostimulatory factors and phagocytosis. Additionally, AE20-SRPL induced the phosphorylation of JNK and inhibition of TLR4 reduced AE20-SRPL-mediated JNK phosphorylation. These results suggested that AE20-SRPL may enhance the production of immunostimulatory factors and phagocytosis through TLR4-dependent activation of JNK in macrophages. Although the present study is limited to in vitro research using a cell model, AE20-SRPL demonstrated potential as a natural material capable of inducing macrophage activation for immune enhancement.

Low pH modulates lipopolysaccharide-induced tumor necrosis factor-alpha expression and macropinocytotic activity in RAW264.7 cells

Inflammation triggers various types of diseases that need to be addressed. Macrophages play important roles in the inflammatory responses. As atherosclerosis progresses, macrophages transform into foam cells. Extracellular acidification is observed at and around bacterial infection and atherosclerotic sites. However, the effects of acidification on the inflammatory response of macrophages and the progression of atherosclerosis have not been fully understood. This study investigates the impact of extracellular acidification on lipopolysaccharide-induced tumor necrosis factor-alpha (TNF-α) expression and macropinocytotic activity in RAW264.7 cells. TNF-α expression is measured by real-time polymerase chain reaction (relative value to glyceraldehyde-3-phosphate dehydrogenase expression). Macropinocytotic activity is measured by neutral red uptake (absorbance at 540 nm). Results show that TNF-α expression increased with decreasing extracellular pH in both un-foamed and foamed cells. Macropinocytotic activity was upregulated at pH 6.8 in un-foamed cells, but downregulated in foamed cells stimulated at low pH. Proton-sensing G protein-coupled receptors (GPCRs) were involved in the expression of TNF-α and in the macropinocytotic activity of foamed cells. In conclusion, this study reveals that extracellular acidification differently affect various inflammatory responses such as LPS-induced TNF-α expression and macropinocytotic activity of RAW264.7 cells and different proton-sensing GPCRs are involved in the different inflammatory responses.

Synthesis and Biological Evaluation of Novel Chlorogenic Acid-Apigenin Conjugates as Anti-acute Gout Agents.

Gout is the second largest metabolic disease worldwide after diabetes, with acute gouty arthritis as most common symptom. Xanthine oxidase (XOD) and the NOD like receptor-3 (NLRP3) inflammasome are the key targets for acute gout treatment. Chlorogenic acid has been reported with a good anti-inflammatory activity, and Apigenin showed an excellent potential in XOD inhibition. Therefore, a series of chlorogenic acid-apigenin (CA) conjugates with varying linkers were designed and synthesized as dual XOD/NLRP3 inhibitors, and their activities both in XOD and NLRP3 inhibition were evaluated. An in vitro study of XOD inhibitory activity revealed that the majority of CA conjugates exhibited favorable XOD inhibitory activity. Particularly, the effects of compounds 10c and 10d, with an alkyl linker on the apigenin moiety, were stronger than that of allopurinol. The selected CA conjugates also demonstrated a favorable anti-inflammatory activity in RAW264.7 cells. Furthermore, compound 10d, which showed the optimal activity both in XOD inhibition and anti-inflammatory, was chosen and its inhibitory ability on NLRP3 and related proinflammatory cytokines was further tested. Compound 10d effectively reduced NLRP3 expression and the secretion of interluekin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) with an activity stronger than the positive control isoliquiritigenin (ISL). Based on these findings, compound 10d exhibits dual XOD/NLRP3 inhibitory activity and, therefore, the therapeutic effects on acute gout is worthy of further study.

Design, Synthesis, and Antirheumatoid Arthritis Mechanism of TLR4 Inhibitors.

A total of 12 carbonyl compounds were synthesized, their lipopolysaccharide induced inhibition, and activity of RAW264.7 cells was evaluated. The most active compound 3k inhibited RAW264.7 cells with IC50 value of 1.02 ± 0.08 μM. Compound 3k significantly inhibited the release of TNF-α, IL-1β, and IL-6 in supernatant for RAW264.7 cells. In vivo collagen-induced arthritis model tests administered orally, compound 3k showed effects similar to those of methotrexate in the positive control group. The preliminary mechanism study showed that compound 3k had an effect on abnormal expression for TLR4, TNF-α, NF-κB protein, and genes related to inflammation signaling pathway in RAW264.7 cells. Meanwhile, compound 3k showed a good affinity for the TLR4 receptor in molecular docking simulation. Therefore, compound 3k may be a promising lead compound for the treatment of rheumatoid arthritis.

Korean Black Goat Extract Exerts Estrogen-like Osteoprotective Effects by Stimulating Osteoblast Differentiation in MC3T3-E1 Cells and Suppressing Osteoclastogenesis in RAW 264.7 Cells.

Postmenopausal osteoporosis, characterized by an imbalance between osteoclast-mediated bone resorption and osteoblast-driven bone formation, presents substantial health implications. In this study, we investigated the role of black goat extract (BGE), derived from a domesticated native Korean goat, estrogen-like activity, and osteoprotective effects in vitro. BGE's mineral and fatty acid compositions were analyzed via the ICP-AES method and gas chromatography-mass spectrometry, respectively. In vitro experiments were conducted using MCF-7 breast cancer cells, MC3T3-E1 osteoblasts, and RAW264.7 osteoclasts. BGE exhibits a favorable amount of mineral and fatty acid content. It displayed antimenopausal activity by stimulating MCF-7 cell proliferation and augmenting estrogen-related gene expression (ERα, ERβ, and pS2). Moreover, BGE positively impacted osteogenesis and mineralization in MC3T3-E1 cells through Wnt/β-catenin pathway modulation, leading to heightened expression of Runt-related transcription factor 2, osteoprotegerin, and collagen type 1. Significantly, BGE effectively suppressed osteoclastogenesis by curtailing osteoclast formation and activity in RAW264.7 cells, concurrently downregulating pivotal signaling molecules, including receptor activator of nuclear factor κ B and tumor necrosis factor receptor-associated factor 6. This study offers a shred of preliminary evidence for the prospective use of BGE as an effective postmenopausal osteoporosis treatment.

Isolation,Identification, Phytochemical Characterization, and In Vitro Anti-Inflammatory Property of an Endophytic Trametes versicolor CL-1 Isolated from Rosa roxburghii.

This study explores the anti-inflammatory potential of an endophytic fungus, Trametes versicolorCL-1, isolated from the fruit tissues of Rosa roxburghii. Morphological and molecular analyses confirmed the identity of CL-1. An ethyl acetate extract (CL-E) from its fermentation broth was subjected to UPLC-HRMS and GNPS molecular networking. The analysis revealed a diverse array of secondary metabolites, including11 terpenes, 7flavonoids, 10cinnamic acid derivatives, 6oligopeptides, and 9fatty acids, as verified by LC-MS/MS. Notably, CL-E exhibited significant in vitro anti-inflammatory activity in RAW264.7 cells. Furthermore, molecular docking studies predicted favorable binding interactions of key compounds 1within CL-E with the NLRP3 inflammasome (PDB ID: 6NPY). These findings suggest T.versicolorCL-1 as a promising source of natural anti-inflammatory agents and unveil R.roxburghiias a potential reservoir for discovering novel bioactive metabolites.

Physicochemical and structural properties of soluble dietary fibres in navel orange peel modified by superfine grinding and their immunomodulatory activities

The physicochemical and structural properties of water soluble dietary fibres (SDF) extracted from navel orange peel modified by superfine grinding were investigated, as well as the immunomodulatory activity in RAW264.7 cells. Results showed that superfine grinding treatment could significantly enhance the extract yield and reduce the particle size of SDF. And the water-holding capacity, swelling capacity, oil-holding capacity, nitrite adsorption capacity and total antioxidant capacity were consequently improved. Modified SDF could significantly decrease the generation of reactive oxygen species (ROS), and promoted the production of NO and some cytokines (IL-1β, TNF-ɑ, MCP-1, and IL-6) in RAW264.7 cells. The expressions of P-P65 and HO-1 in SDF-M200 and SDF-M500 were significantly up-regulated, suggesting that SDF could mildly induce macrophage activation and enhance the immune activity. These indicated that SDF modified by superfine grinding could serve as a promising candidate of immunomodulatory ingredient for functional foods.

The antioxidant activity and metabolomic analysis of the supernatant of Streptococcus alactolyticus strain FGM

Strain-specific probiotics can present antioxidant activity and reduce damage caused by oxidation. Streptococcus alactolyticus strain FGM (S. alactolyticus strain FGM) isolated from the chicken cecum shows potential probiotic properties which have been previously demonstrated. However, the antioxidant properties of S. alactolyticus strain FGM remain unknown. In this view, cell-free supernatant (CFS), intact cells (IC) and intracellular extracts (CFE) of strain FGM and 3 strains of Lactobacillus (LAB) were prepared, and their scavenging capacities against DPPH, hydroxyl radicals and linoleic acid peroxidation inhibitory were compared in this study. The effects of strain FGM cell-free supernatant (FCFS) on NO production, activity of SOD and GSH-Px in RAW264.7 cells and LPS-induced RAW264.7 cells were analyzed. The metabolites in the supernatant were quantitated by N300 Quantitative Metabolome. It was shown that the physicochemical characteristics of CFS to scavenge DPPH, hydroxyl radicals, and linoleic acid peroxidation inhibitory were significantly stronger than that of IC and CFE in the strain FGM (P < 0.05), respectively 87.12% ± 1.62, 45.03% ± 1.27, 15.63% ± 1.34. FCFS had a promotional effect on RAW264.7 cells, and significantly elevated SOD and GSH-Px activities in RAW264.7 cells. 25 μL FCFS significantly promoted the proliferation of RAW264.7 cells induced by LPS, increased the activities of SOD and GSH-PX, and decreased the release of NO. Furthermore, among the differential metabolites of FCFS quantified by N300, 12 metabolites were significantly up-regulated, including lactic acid, indole lactic acid, linoleic acid, pyruvic acid etc., many of which are known with antioxidant properties. In conclusion, FCFS had good antioxidant properties and activity, which can be attributed to metabolites produced from strain FGM fermentation. It was further confirmed that S. alactolyticus strain FGM and its postbiotic have potential probiotic properties and bright application prospects in livestock and poultry breeding.

Immune Activity in RAW264.7 Macrophages through MAPKs and NF-ĸB Signaling Pathway of Low-Temperature Extracts of Allomyrina dichotoma Larvae

Immune Activity in RAW264.7 Macrophages through MAPKs and NF-ĸB Signaling Pathway of Low-Temperature Extracts of <i>Allomyrina dichotoma</i> Larvae

Antimicrobial Activities against Skin Flora from Tart-cherry Extracts (Prnus cerasus)

Purpose: In this study, antibacterial and anti-oxidant activities of Tart-cherry hydrothermal extracts (TCW) and ethanol extracts (TCE) against dermatological bacteria were measured, and the cell viability and anti-inflammatory effects were investigated in RAW264.7 macrophages to determine their characteristics for use as functional cosmetic materials. Methods: Anti-oxidant activity was determined using the DPPH method. The antibacterial activity against dermatophilosis was confirmed by the size of the clear zone formed by DISK diffusion and MIC measurement. The activity against an antibiotic-resistant MRSA strain was also measured. Anti-inflammatory response was identified by measuring cytotoxicity and NO activity in RAW264.7 macrophages, in which inflammatory reactions were induced with LPS. Results: Anti-oxidant activity of the TCE was 85.6%, and of TCW was 88.4% at 500 μg/mL. The size of the clear zone was confirmed using Cutibacterium acnes (18 mm), &lt;i&gt;Staphylococcus epidermis&lt;/i&gt; (&lt;i&gt;S. epidermis&lt;/i&gt; ) (4 mm), and &lt;i&gt;Staphylococcus aureus&lt;/i&gt; (&lt;i&gt;S. aureus&lt;/i&gt; ) (16 mm). No antifungal activity against &lt;i&gt;Candida albicans&lt;/i&gt; was observed. RAW264.7 macrophages showed a survival rate of &gt;90% at &lt;250 μg/mL of extract. Additionally, a concentration-dependent inhibition of NO production was observed. Conclusion: Tart-cherry extracts possessed high anti-oxidant activity and anti-inflammatory efficacy. In addition to the antibacterial activity against the skin-residing bacteria, &lt;i&gt;S. aureus&lt;/i&gt; and &lt;i&gt;S. epidermis&lt;/i&gt;, high activity against the acne-causing strain &lt;i&gt;C. acnes&lt;/i&gt; suggests a sufficient potential of Tart-cherry extracts as a raw material for the preparation of cosmetics and bioproducts.

Penehyclidine hydrochloride alleviates LPS-induced inflammatory responses and oxidative stress via ROS/Nrf2/HO-1 activation in RAW264.7 cells.

Inflammation is a biological response of the immune system to harmful stimuli. Penehyclidine hydrochloride (PCH) can alleviate inflammation and oxidative stress by activating reactive oxygen species (ROS), nuclear factor erythroid 2-related factor (Nrf2) and heme oxygenase 1 (HO-1) in animal models, but there is a lack of cellular evidence. This study investigated the effects of PHC on lipopolysaccharide (LPS)-induced inflammation response and oxidative stress in RAW264.7 cells. RAW264.7 cells were treated with 1 μg/mL or 5 μg/mL of PHC, with interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-α), IL-1β, and prostaglandin E2 (PGE2) levels measured with enzyme-linked immunosorbent assay (ELISA) and nitric oxide (NO) measured using the Griess test. Reactive oxygen species were examined with flow cytometry and immunofluorescence, and b-related factor 2 (BRF-2) and NAD(P)H-quinone oxidoreductase 1 (NQO1) using western blot. Penehyclidine hydrochloride partly, but substantially, reversed LPS-related NO and PGE2 production by RAW264.7 cells in a dose-dependent manner and suppressed LPS-induced expression of IL-6, TNF-α and IL-1β messenger ribonucleic acid (mRNA), secretion of IL-6, TNF-α and IL-1β, and ROS production. Lipopolysaccharide stimulation did not affect Nrf2, heme oxygenase 1 (HO-1) or NQO1 protein expression in RWA264.7 cells not treated with PHC. However, PHC treatment significantly elevated Nrf2, HO-1 and NQO1 protein in LPS-treated RWA264.7 cells, an effect that was dose-dependent. The ROS scavenging using N-acetyl-L-cysteine abolished the PHC-induced upregulation of Nrf2 and HO-1. Penehyclidine hydrochloride may alleviate LPS-induced inflammation and oxidative stress by activating Nrf2 signaling in RAW264.7 macrophages. These findings suggest that PHC could alleviate inflammation by targeting activated macrophages.

Insights into the mechanism of action of pterostilbene against influenza A virus-induced acute lung injury

BackgroundSevere respiratory system illness caused by influenza A virus infection is associated with excessive inflammation and abnormal apoptosis in alveolar epithelial cells (AEC). However, there are limited therapeutic options for influenza-associated lung inflammation and apoptosis. Pterostilbene (PTE, trans-3,5-dimethoxy-4-hydroxystilbene) is a dimethylated analog of resveratrol that has been reported to limit influenza A virus infection by promoting antiviral innate immunity, but has not been studied for its protective effects on virus-associated inflammation and injury in AEC. PurposeOur study aimed to investigate the protective effects and underlying mechanisms of PTE in modulating inflammation and apoptosis in AEC, as well as its effects on macrophage polarization during influenza virus infection. Study design and methodsA murine model of influenza A virus-mediated acute lung injury was established by intranasal inoculation with 5LD50 of mouse-adapted H1N1 viruses. Hematoxylin and eosin staining, immunofluorescence, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, western blotting, Luminex and flow cytometry were performed. ResultsPTE effectively mitigated lung histopathological changes and injury induced by H1N1 viruses in vivo. These beneficial effects of PTE were attributed to the suppression of inflammation and apoptosis in AEC, as well as the modulation of M1 macrophage polarization. Mechanistic investigations revealed that PTE activated the phosphorylated AMP-activated protein kinase alpha (P-AMPKα)/sirtui1 (Sirt1)/PPARγ coactivator 1-alpha (PGC1α) signal axis, leading to the inhibition of nuclear factor kappa-B (NF-κB) and p38 mitogen-activated protein kinase (MAPK) signaling induced by H1N1 viruses, thereby attenuating inflammation and apoptosis in AEC. PTE also forced activation of the P-AMPKα/Sirt1/PGC1α signal axis in RAW264.7 cells, counteracting the activation of phosphorylated signal transducer and activator of transcription 1 (P-STAT1) induced by H1N1 viruses and the augment of P-STAT1 activation in RAW264.7 cells with interferon-gamma (IFN-γ) pretreatment before viral infection, thereby reducing H1N1 virus-mediated M1 macrophage polarization as well as the enhancement of macrophages into M1 phenotypes elicited by IFN-γ pretreatment. Additionally, the promotion of the transition of macrophages towards the M2 phenotype by PTE was also related to activation of the P-AMPKα/Sirt1/PGC1α signal axis. Moreover, co-culturing non-infected AEC with H1N1 virus-infected RAW264.7 cells in the presence of PTE inhibited apoptosis and tight junction disruption, which was attributed to the suppression of pro-inflammatory mediators and pro-apoptotic factors in an AMPKα-dependent manner. ConclusionIn conclusion, our findings suggest that PTE may serve as a promising novel therapeutic option for treating influenza-associated lung injury. Its ability to suppress inflammation and apoptosis in AEC, modulate macrophage polarization, and preserve alveolar epithelial cell integrity highlights its potential as a therapeutic agent in influenza diseases.

Purification and Structural Characterization of Polysaccharides from Polygonum multiflorum Thunb. and Their Immunostimulatory Activity in RAW264.7 Cells.

Polygonum multiflorum Thunb. (PM) and derived products are broadly utilized in Chinese traditional medicine. According to our previous research, PM mostly contains polysaccharides, which display a wide range of biological activities. Two water-soluble polysaccharides (PMPs-1 and PMPs-2) were obtained from PM by DEAE-Cellulose and Sephadex G-100 column chromatography. Colorimetry, HPGPC-MALLS-RID, HPLC-PDA, methylation, FT-IR, NMR, and SEM were used to characterize these polysaccharides. PMPs-1 and PMPs-2 had average molecular weights of 255.5 and 55.7 kDa, respectively. PMPs-1 consisted of Man, Glc, Gal, and Ara at 0.9:78.6:1.0:1.6 and was a glucan with → 4)-Glcp-(1 → as a backbone. Meanwhile, PMPs-2, an acidic polysaccharide, comprised Rha, GalA, Glc, Gal, and Ara at 3.2:20.3:2.7:1.0:8.3. PMPs-1 and PMPs-2 significantly improved the proliferation of RAW 264.7 cells and induced NO, TNF-α, and IL-6 release. This study reveals that these two polysaccharides can be explored as novel immunomodulators and provide a basis for further development of PM in food and pharmaceutical industries.

Isoliquiritigenin reduces experimental autoimmune prostatitis by facilitating Nrf2 activation and suppressing the NLRP3 inflammasome pathway

BackgroundChronic prostatitis and chronic pelvic pain syndrome (CP/CPPS) lead to severe irritation and impaired sperm quality in males. However, current therapeutic options often fail to achieve satisfactory effects. Consequently, the investigation of novel treatment strategies or remedies holds substantial clinical importance. As a flavonoid monomer, isoliquiritigenin (ISL) has been shown to possess anti-inflammatory activity, especially in several chronic nonspecific-inflammatory conditions. Thus, an exploration of the possible anti-inflammatory effects of ISL on CP/CPPS, a chronic aseptic inflammation of the prostate, has significant potential. MethodsAn experimental autoimmune prostatitis (EAP) model was used for the evaluation of the anti-inflammatory effects of ISL. It was found that ISL treatment could reduce the secretion and invasion of pro-inflammatory cytokines in prostate tissue. In EAP mice, ISL treatment also reduced oxidative stress (OS) and activation of the NLRP3 inflammasome. In vitro, ISL upregulated the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and inhibited NLRP3 inflammasome activation in RAW264.7 macrophages exposed to lipopolysaccharide (LPS). ResultsTreatment with ISL treatment relieved prostate inflammation and pelvic pain in EAP mice. Both in vivo and in vitro, ISL treatment activated Nrf2/HO-1 signaling, which in turn inhibited oxidative stress and activation of the NLRP3 inflammasome. Blockade of Nrf2/HO-1 signaling abolished the inhibitory effects of ISL on oxidative stress and NLRP3 inflammasome activation. ConclusionsIsoliquiritigenin reduced experimental autoimmune prostatitis by facilitating Nrf2 activation and suppressing the NLRP3 inflammasome pathway.

Development of gold nanocluster complex for the detection of tumor necrosis factor-alpha based on immunoassay

Tumor necrosis factor-alpha, TNF-α, a cytokine recognized as a key regulator of inflammatory responses, is primarily produced by activated monocytes and macrophages. Measuring TNF-α levels serves as a valuable indicator for tracking several diseases and pathological states. Gold nanotechnology has been identified as a highly effective catalyst with unique properties for measuring inflammatory cytokines. This study aimed to synthesize gold nanoclusters (AuNCs) and the AuNCs-streptavidin system, along with their characterizations and spherical morphology. The detection of TNF-α antigen with AuNCs was determined, and a new immunoassay-based AuNCs analytical platform was studied. In this study, it was demonstrated that the synthesized AuNCs and AuNCs-streptavidin showed a bright-yellow appearance with absorption peaks at A600 and A610 nm, respectively. The approximately spherical shape was observed by TEM analysis. The AuNCs demonstrated a sensitivity limit for the detection of the TNF-α antigen, with a linear dose-dependent detection range of less than 1.25 ng/mL. The products of the band sizes and band intensities were proportional to the amount of TNF-α in the range of ∼80 kDa, ∼55 kDa, and ∼ 25 kDa in western blot analysis. The TNF-α in cell lysate was successfully detected using an immunoassay after the activation of RAW264.7 cells with lipopolysaccharide (LPS). This assay may serve as a viable alternative for TNF-α detection with high speed, sensitivity, and qualities, ensuring its broad applications.

Structural characterization of polysaccharides from Dictyophora rubrovolvata mycelium and their immunostimulatory activity in RAW264.7 cells

In this study, a homogeneous polysaccharide, named, DRMP-1 was isolated from the mycelium of Dictyophora rubrovolvata by water extraction and alcohol precipitation, and its structure and activity were studied. The molecular weight of DRMP-1 is 2.33 × 106 Da, and it does not contain protein and nucleic acid. DRMP-1 is composed of mannose, glucose and galactose with a molar ratio of 0.04: 1: 0.04. Through methylation and NMR analysis, DRMP-1 is mainly composed of 1 → 6)-α-Galp-( 1 →, T-α-D-Glcp, →1)-α-D-Manp, → 6)-β-D-Glcp-( 1 →, → 4, 6)- α-D-Manp-( 1 →, → 4)- α-D-Glcp-( 1 →, → 3)-β-D-Glcp-( 1 → and → 4, 6)- α-D-Glcp-( 1 →. In addition, the immunomodulative activity of DRMP-1 was evaluated by establishing the RAW264.7 cell model. The results showed that DRMP-1 could significantly increase phagocytosis activity and NO production more than twice as much as that of the blank group, and could increase intracellular ACP, LSZ and SOD activities. Therefore, it is concluded that the polysaccharide of Dictyophora rubrovolvata mycelium may be a potential immunomodulator.

Pharmacological activities of extracts from different parts of &lt;i&gt;Cirsium japonicum&lt;/i&gt; var &lt;i&gt;spinossimum&lt;/i&gt;

Purpose: To evaluate the antioxidant, anti-inflammatory, inhibitory and cell proliferation activities of Cirsium japonicum (C. japonicum) var. spinossimum.&#x0D; Methods: Phytochemicals (polyphenols, flavonoids, and silymarin) in leaves, roots, and seeds of C. japonicum var. spinossimum were quantified using high-performance liquid chromatography (HPLC). Antioxidant activity was investigated on 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals. Anti-inflammatory activity was investigated on nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW264.7 cells. Immunostimulant and hair loss prevention was confirmed through 3'-{1-((phenylamino)-carbonyl)-3, 4-tetrazolium}bis(4-methoxy-6-nitro)benzenesulfonic acid hydrate (XTT)-based cell proliferation assays using Jurkat cells and human follicle dermal papilla cells (HFDPC).&#x0D; Results: Extract from seeds exhibited the highest total polyphenol content (610.08 ± 32.40 mg GAE/g), as well as the highest total flavonoid content, while the total polyphenol content of leaves and root extracts were similar. The RC50 values for seed extract against ABTS and DPPH radicals were 8.20 ± 0.02 µg/mL and 40.13 ± 2.47 µg/mL, respectively. In particular, all six Silymarin derivatives were found to be highest in the seed extract. The C. japonicum var. spinossimum seed extract (CSE) showed significant (p &lt; 0.05) nitric oxide (NO) inhibitory activity in RAW264.7 cells induced by LPS at all concentrations (12.5, 25, 50, 100 and 200 µg/mL) and exhibited significant (p &lt; 0.05) cell proliferation activity in Jurkat and hair follicle dermal papilla cells (HFDPC) at 25 µg/mL.&#x0D; Conclusion: Extract of C. japonicum var. spinossimum seeds contain polyphenols and flavonoids and show significant NO inhibitory and cell proliferation activity in Jurkat and HFDPC. Therefore, Cirsium japonicum is a promising source of anti-inflammatory and anticancer agents.

Discovery of bi-4-methoxycarbonyl-2-quinolone and evaluation of its anti-inflammatory and anti-cancer activity in vitro

A novel alkaloid with a hexa-tetra-hexa-cyclic skeleton, Bi-4-methoxycarbonyl-2-quinolone (1), was discovered during the investigation of Brucea javanica. Additionally, six known alkaloids (2–7) were also found. The chemical structures of these compounds were identified using HRESIMS and 1D and 2D NMR spectroscopic analysis. Additionally, the absolute configuration of the new compound 1 was determined through X-ray single crystal diffraction. Compound 1 exhibited significant anti-inflammatory activity in RAW264.7 cells and demonstrated promising anti-cancer effects in Lewis cells.