Pro-inflammatory Cytokines In RAW264 Research Articles

Establishment of a workflow for high-throughput identification of anti-inflammatory peptides from sea cucumbers

Developing an effective workflow for screening anti-inflammatory peptides is crucial for discovering novel food-derived anti-inflammatory peptides and optimizing the screening and identification process of bioactive peptides. Virtual screening identified three major yolk proteins as target precursor proteins for anti-inflammatory peptides in sea cucumbers. A portfolio of 170 peptides was identified from hydrolysates after 9 h of alcalase treatment by combining antioxidant activity determination and peptidomics analysis. Among these, 12 high-confidence anti-inflammatory peptides were identified through virtual screening. Three of these peptides were shown to effectively inhibit the production of NO and the release of pro-inflammatory cytokines in RAW264.7 cells. Molecular docking demonstrated that these three peptides exerted their anti-inflammatory effects primarily by binding to the active sites of cyclooxygenase-2 and inducible nitric oxide synthase through hydrophobic interactions. This study provided a reference workflow for screening anti-inflammatory peptides, facilitating the discovery of novel anti-inflammatory peptides and the high-value utilization of sea cucumber cooking liquid.

Spatial distribution of antioxidant activity in baguette and its modulation of proinflammatory cytokines in RAW264.7 macrophages

Baguette is a globally acclaimed bakery staple, composed by a crispy crust and soft crumb, both containing Maillard reaction products (MRPs) with potential bioactivities. However, MRPs’ impacts on the nutritional and health attributes of baguette, particularly in terms of cellular and biological functions, are yet to be clearly elucidated. This study chemically characterizes the crust and crumb of baguettes and investigates the influence of the Maillard reaction on baguette’s nutritional profile, especially in the antioxidant and anti-inflammatory effects. The findings indicate an increase in browning intensity and advanced glycation end products (AGEs) from the baguette’s interior to its exterior, alongside a significant rise in the antioxidant capacity of the crust, suggesting the Maillard reaction’s role in boosting antioxidative properties. Both the crust and crumb demonstrated strong cytocompatibility with immune cells, capable of reducing cellular oxidative stress and regulating intracellular free radical levels. The crust effectively countered peroxyl radical-induced cell membrane hyperpolarization by 91% and completely neutralized the suppression of oxygen respiration in mitochondria, displaying higher efficacy than the crumb. In contrast, crumb extracts were more potent in inhibiting lipopolysaccharide-induced expression of proinflammatory cytokines, such as interleukins-1β (IL-1β) and IL-6, in macrophages. It could provide the fundamental data and cell-based approach for investigating the biological impacts of bread on immune responses, contributing to the refinement and supplementation of nutritional recommendations.

Biological and chemical enhancements of Elaeocarpus sylvestris var. ellipticus through fermentation: Implications for therapeutic and industrial applications

This study aimed to evaluate the biological and chemical improvements that take place during the yeast fermentation of hot water-extracted Elaeocarpus sylvestris (HES). HES was diluted with sterile distilled water to a final concentration of 1:200 (v/v), followed by the inoculation of Pichia fermentans (PF) and Saccharomyces cerevisiae (SC) were inoculated as starters. The fermentation by yeast strains proceeded for five days, during which the viable cell count and pH were monitored. The radical scavenging activity was assessed using DPPH and ABTS assays. The impact of HES on proinflammatory cytokines in RAW 264.7 cells and cytotoxicity in HT-29, MCF-7, and INT407 cells was examined. Changes in metabolic profile during fermentation were analyzed using GC-TOF-MS and UPLC-Orbitrap-MS/MS. HES fermented with PF and SC maintained viable cell counts throughout the 5-day fermentation period. The pH levels were consistently within the range of 3.62 ± 0.03 to 3.65 ± 0.15. The expression of TNF-α and IL-6 significantly decreased at all concentrations (31–500 mg/mL) in RAW cells treated with lipopolysaccharide and HES, with cytokine expression levels returning to baseline levels. The cell viability of INT407 significantly increased following HES treatment (p < 0.05). Fermentation of HES with PF or SC led to significant changes in the composition ratios of carboxylic acids and derivatives, and organooxygen compounds. The HES extract demonstrates potential applicability as a raw material with antioxidative, anti-inflammatory, and anticancer properties for applications in the food and cosmetic industries.

The beneficial effects of Rosuvastatin in inhibiting inflammation in sepsis.

Microbial infection-induced sepsis causes excessive inflammatory response and multiple organ failure. An effective strategy for the treatment of sepsis-related syndromes is still needed. Rosuvastatin, a typical β-hydroxy β-methylglutaryl-CoA reductase inhibitor licensed for reducing the levels of low-density lipoprotein cholesterol in patients with hyperlipidemia, has displayed anti-inflammatory capacity in different types of organs and tissues. However, its effects on the development of sepsis are less reported. Here, we found that the administration of Rosuvastatin reduced the mortality of sepsis mice and prevented body temperature loss. Additionally, it inhibited the production of inflammatory cytokines such as tumor necrosis factor (TNF-α), Interleukin-6 (IL-6), interleukin-1β (IL-1β), and migration inhibitory factor (MIF) in peritoneal lavage supernatants of animals. The increased number of mononuclear cells in the peritoneum of sepsis mice was reduced by Rosuvastatin. Interestingly, it ameliorated lung inflammation and improved the hepatic and renal function in the sepsis animals. Further in vitro experiments show that Rosuvastatin inhibited lipopolysaccharide (LPS)-induced production of proinflammatory cytokines in RAW 264.7 macrophages by preventing the activation of nuclear factor kappa-B (NF-κB). Our findings demonstrate that the administration of Rosuvastatin hampered organ dysfunction and mitigated inflammation in a relevant model of sepsis.

Selegiline modulates inflammatory indicators in RAW 264.7 macrophages and LPS-aggravated CFA-induced rheumatoid arthritis in rats

Introduction and aim. Rheumatoid arthritis (RA) causes pain, inflammation, and deformities in numerous joints. Monoamine oxidase B (MOA-B) inhibitor selegiline exhibits anti-inflammatory characteristics and has the propensity to scavenge free radicals. Therefore, the aim of this research comprises of assessing the effect of selegiline on proinflammatory cytokines in RAW 264.7 macrophages as well as its capacity to improve various arthritic parameters in rats with lipopolysaccharide (LPS) accelerated complete Freund’s adjuvant (CFA) induced RA. Material and methods. In RAW 264.7 cells (lipopolysaccharide accelerated), nitric oxide (NO), tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), inducible nitric oxide synthase (iNOS), and prostaglandin E2 (PGE2 ) were determined after treat ment with selegiline. Different arthritic parameters were analyzed after administration of selegiline in LPS accelerated CFA-induced arthritis in rats. Results. LPS escalates NO, TNF-α, IL-6, iNOS, and PGE2 quantities in the RAW 264.7 cells, which was minimized by selegiline at 100 µg/mL and 150 µg/mL respectively. In rats, CFA induction causes a decrease in body weight, elevation of paw volume, splenic index, and arthritic index, which are further accelerated by LPS. 20 mg/kg of selegiline managed all these arthritic parameters effectively, including TNF-α, IL-6, and a few other biochemical parameters. Conclusion. Selegiline may be beneficial in RA extenuating joint and cartilage damage, and modulating inflammatory responses.

VX-702 Ameliorates the Severity of Sepsis-Associated Acute Kidney Injury by Downregulating Inflammatory Factors in Macrophages.

Sepsis-associated acute kidney injury (S-AKI) contributes to high mortality, but it is lack of specific treatments. We aimed to investigate the underlying mechanism of S-AKI and to identify target drugs to alleviate AKI. We establish a stable mouse model of S-AKI by Pseudomonas aeruginosa incision infection. Based on high-throughput sequencing and bioinformatics analysis, we investigated the underlying mechanism and selected the target drug (VX-702) for S-AKI. An in vitro model established by co-cultured of kidney tubular epithelial cell line (TCMK-1) cells with lipopolysaccharide (LPS)-induced leukemic monocyte/macrophage cells (RAW264.7), we explored the effect of VX-702 on S-AKI. The data showed interleukin (IL)-6 and IL-1β were the hub genes, and the mitogen-activated protein kinase (MAPK) signaling pathway was the main pathway involved in S-AKI. Administration of VX-702 by oral gavage decreased the elevated concentrations of IL-6, IL-1β, serum creatinine, and blood urea nitrogen in mice with S-AKI. Moreover, VX-702 reduced the number of apoptotic cells in damaged kidney tissues. Cell viability was decreased, and the number of apoptotic cells was increased in TCMK-1 cells co-cultured with LPS-induced RAW264.7 cells compared to LPS-induced TCMK-1 cells. VX-702 treatment reversed this effect. VX-702 treatment reduced the levels of phosphorylated p38 MAPK and proinflammatory cytokines in RAW264.7 cells and the supernatant. VX-702 could bind IL-6, IL-1β and MAPK, and affect the binding of IL-1β and its receptor, as demonstrated by molecular docking. VX-702 ameliorated S-AKI by inhibiting the release of proinflammatory cytokines from macrophages, indicating its potential as a novel therapeutic for S-AKI treatment.

Nicotinic acetylcholine receptor-mediated effects of varenicline on LPS-elevated prostaglandin and cyclooxygenase levels in RAW 264.7 macrophages.

Introduction: The purpose of this study is to delineate anti-inflammatory and antioxidant potential of varenicline, a cigarette smoking cessation aid, on decreasing lipopolysaccharide (LPS)-elevated proinflammatory cytokines in RAW 264.7 murine macrophage cultures which we showed earlier to occur via cholinergic anti-inflammatory pathway (CAP) activation. To this end, we investigated the possible suppressive capacity of varenicline on LPS-regulated cyclooxygenase (COX-1 and COX-2) via α7 nicotinic acetylcholine receptor (α7nAChR) activation using the same in vitro model. Materials and Methods: In order to test anti-inflammatory effectiveness of varenicline, the levels of COX isoforms and products (PGE2, 6-keto PGF1α, a stable analog of PGI2, and TXA2) altered after LPS administration were determined by Enzyme Linked Immunosorbent Assay (ELISA). The antioxidant effects of varenicline were assessed by measuring reductions in reactive oxygen species (ROS) using a fluorometric intracellular ROS assay kit. We further investigated the contribution of nAChR subtypes by using non-selective and/or selective α7nAChR antagonists. The results were compared with that of conventional anti-inflammatory medications, such as ibuprofen, celecoxib and dexamethasone. Results: Varenicline significantly reduced LPS-induced COX-1, COX-2 and prostaglandin levels and ROS to an extent similar to that observed with anti-inflammatory agents used. Discussion: Significant downregulation in LPS-induced COX isoforms and associated decreases in PGE2, 6-keto PGF1α, and TXA2 levels along with reduction in ROS may be partly mediated via varenicline-activated α7nAChRs.

GehB Inactivates Lipoproteins to Delay the Healing of Acute Wounds Infected with Staphylococcus aureus.

Staphylococcus aureus is one of the most prevalent bacteria found in acute wounds. S. aureus produces many virulence factors and extracellular enzymes that contribute to bacterial survival, dissemination, and pathogenicity. Lipase GehB is a glycerol ester hydrolase that hydrolyzes triglycerides to facilitate the evasion of S. aureus from host immune recognition. However, the role and mechanism of lipase GehB in skin acute wound healing after S. aureus infection remain unclear. In this study, we found that the gehB gene deletion mutant (USA300ΔgehB) stimulated significantly higher levels of pro-inflammatory cytokines in RAW264.7 and Toll-like receptor 2 (TLR2)-transfected HEK293 cells than the wild-type USA300 strain did. Recombinant GehB-His treated lipoprotein (Lpp) reduced stimulation of TLR2-dependent TNF-α production by RAW264.7 macrophages. GehB delayed the skin acute wound healing in BALB/c mice infected with S. aureus, while wound healing was similar in C57BL/6 TLR2-/- mice infected with either wild-type USA300 or USA300ΔgehB. In BALB/c mice,we also observed more bacterial survival, less leukocyte recruitment, lower IL-8 production, and adipocyte differentiation in USA300-infected skin acute wound tissues than those in USA300ΔgehB-challenged ones. Our data indicated that GehB inactivates lipoproteins to shield S. aureus from innate immune killing, resulting in delayed the healing of skin acute wounds infected with S. aureus.

Discovery and verification of anti-inflammatory-related quality markers in the aerial part of Bupleurum scorzonerifolium by UPLC-Q-TOF-MS/MS and in RAW 264.7 cells and a zebrafish model.

The root of Bupleurum scorzonerifolium Willd. (BS) is officially recognized in the Chinese Pharmacopoeia. In contrast, the aerial part of BS (ABS), accounting for 80% of BS, is typically discarded, causing potential waste of medicinal resources. ABS has shown benefits in the treatment of inflammation-related diseases in China and Spain, and the material basis underlying its anti-inflammatory effects must be systematically elucidated for the rational use of ABS. We aimed to screen and validate the anti-inflammatory quality markers (Q-markers) of ABS and to confirm the ideal time for ABS harvesting. The chemical components and anti-inflammatory effects of ABS from 10 extracted parts were analyzed by UPLC-Q-TOF-MS/MS and in a lipopolysaccharide (LPS)-induced cell model. Anti-inflammatory substances were screened by Pearson bivariate analysis and gray correlation analysis, and the anti-inflammatory effects were verified in a zebrafish tail-cutting inflammation model. HPLC was applied to measure the Q-marker contents of ABS in different harvesting periods. Ten ABS extracts effectively alleviated the increase in LPS-induced proinflammatory cytokines in RAW 264.7 cells. Forty components were identified from them, among which 27 were common components. Eight components were correlated with anti-inflammatory effects, which were confirmed to reverse the expression of proinflammatory and anti-inflammatory factors in a zebrafish model. Chlorogenic acid, hypericin, rutin, quercetin, and isorhamnetin can be detected by HPLC, and the maximum contents of these five Q-markers were obtained in the sample harvested in August. The anti-inflammatory Q-markers of ABS were elucidated by chromatographic-pharmacodynamic-stoichiometric analysis, which served as a crucial basis for ABS quality control.

Transcriptome analysis reveals modulation of differentially expressed genes in LPS-treated mouse macrophages (RAW264.7cells) by grouper (Epinephelus coioides) Epinecidin-1.

The marine antimicrobial peptide Epinecidin (Epi)-1 has been shown to exert direct antimicrobial and immunomodulatory actions in teleost, mammalian and avian organisms. For instance, Epi-1 can suppress bacterial endotoxin lipolysachcharide (LPS)-induced proinflammatory cytokines in RAW264.7 murine macrophages. However, it remains unknown how Epi-1 might broadly affect non-activated and LPS-activated macrophages. To address this question, we performed a comparative transcriptomic analysis of non-treated and LPS-treated RAW264.7cells in the presence and absence of Epi-1. Gene enrichment analysis was conducted on filtered reads, followed by GO and KEGG analyses. The results showed that Epi-1 treatment modulated pathways and genes associated with nucleoside binding, intramolecular oxidoreductase activity, GTPase activity, peptide antigen binding, GTP binding, ribonucleoside/nucleotide binding, phosphatidylinositol binding and phosphatidylinositol-4-phosphate binding. Based on the GO analysis results, we performed real-time PCR at different treatment times to compare expression levels of selected proinflammatory cytokines, anti-inflammatory cytokines, MHC, proliferation and differentiation genes. Epi-1 decreased expression of the proinflammatory cytokines, TNF-α, IL-6 and IL-1β, and it increased the anti-inflammatory cytokine TGFβ and Sytx1. MHC-associated genes, GM7030, Arfip1, Gpb11 and Gem, were induced by Epi-1, which is expected to enhance the immune response against LPS. Immunoglobulin-associated Nuggc was also upregulated by Epi-1. Finally, we found that Epi-1 downregulated the expression of host defense peptides CRAMP, Leap2 and BD3. Taken together, these findings suggest that Epi-1 treatment induces orchestrated changes in the transcriptome of LPS-stimulated RAW264.7cells.

Immunomodulatory Effect of Benincasa hispida Extract Fermented by Bacillus subtilis CJH 101 on RAW 264.7 Macrophages

This study aimed to confirm the immunomodulatory effects of fermented Benincasa hispida extract (HR1901-BS) on RAW 264.7 macrophages. B. hispida was fermented for 72 h using Bacillus subtilis CJH 101. To evaluate the efficacy of HR1901-BS in enhancing macrophage function, we measured and compared the levels of macrophage activation-related markers in HR1901-BS-stimulated RAW 264.7 cells. We evaluated the effects on nitric oxide (NO) production and the release of pro-inflammatory cytokines (interleukin IL-1β, IL-6, and tumor necrosis factor TNF-β) in the RAW 264.7 cell line. We confirmed that HR1901-BS affected macrophage activation by inducing a significantly high dose-dependent NO production and increasing the release of pro-inflammatory cytokines in RAW 264.7 macrophages. It also suggested that the immunomodulatory effect by HR1901-BS occurred mainly as a result of the activation of cyclooxygenase-2 (COX-2), inducible NO synthase (iNOS), and mitogen-activated protein kinase (MAPK)/nuclear factor-κB (NF-κB) pathways. Our results indicate that HR1901-BS is a promising candidate as a functional material that enhances immune responses through macrophage activation.

Hydraulic calcium silicate-based root canal sealers mitigate proinflammatory cytokine synthesis and promote osteogenesis in vitro

Background/purposeThe mineralized tissue-inductive ability and anti-inflammatory properties of hydraulic calcium silicate-based (HCSB) sealers have not been fully elucidated. This study aimed to evaluate the effects of the HCSB sealers Bio-C sealer (BioC), Well-Root ST (WST), and EndoSequence BC sealer (BC), on osteoblastic differentiation/mineralization and proinflammatory cytokine synthesis by macrophages. Materials and methodsDiluted extracts of set sealers or calcium chloride solutions of approximately equivalent Ca2+ concentrations were applied to a mouse osteoblastic cell line (Kusa-A1 cells) and lipopolysaccharide-stimulated mouse macrophage cell line (RAW264.7 cells). Expressions of osteoblastic markers in Kusa-A1 cells and proinflammatory cytokines in RAW264.7 cells were evaluated by reverse transcription-quantitative polymerase chain reaction and enzyme-linked immunosorbent assays. Mineralized nodules were detected by Alizarin red S staining. Cell proliferation was assessed by WST-8 assay and cell attachment on set sealers was examined by scanning electron microscopy. ResultsThe three sealer extracts significantly upregulated osteocalcin and osteopontin mRNA, and promoted significant mineralized nodule formation in Kusa-A1 cells. The three sealer extracts significantly downregulated the mRNA expressions of interleukin (IL)-1α, IL-1β, IL-6, and tumor necrosis factor (TNF)-α and protein levels of IL-6 and TNF-α in RAW264.7 cells. Calcium chloride solutions induced osteoblastic differentiation/mineralization. AH Plus Jet (a control sealer) extract did not. The three HCSB sealers did not interfere with the growth and attachment of Kusa-A1 cells. ConclusionBioC, WST, and BC were biocompatible, upregulated osteoblastic differentiation/mineralization, and downregulated proinflammatory cytokine expression. Ca2+ released from HCSB sealers might be involved, at least in part, in the induction of osteoblastic differentiation/mineralization.

Potent Anti-Inflammatory Effects of a Helix-to-Helix Peptide against Pseudomonas aeruginosa Endotoxin-Mediated Sepsis.

Although considerable scientific research data is available for sepsis and cytokine storm syndrome, there is a need to develop new treatments or drugs for sepsis management. Antimicrobial peptides (AMPs) possess anti-bacterial and anti-inflammatory activity, neutralizing toxins such as lipopolysaccharides (LPS, endotoxin). Most AMPs have been designed as a substitute for conventional antibiotics, which kill drug-resistant pathogens. The present study aimed to determine the anti-inflammatory potential of 10 designed XIW (X: lysine, arginine, or glutamic acid) α-helical peptides in macrophages and a mouse model in the presence of LPS. Among them, WIKE-14, a peptide with a helix-to-helix structure, having the 12th amino acid substituted with glutamic acid, suppressed pro-inflammatory cytokines in RAW 264.7 macrophages. This reaction was mediated by the inhibition of the binding between LPS and macrophages. In addition, the WIKE-14 peptide exhibited a potent anti-inflammatory activity in mice ears and lungs inflamed using LPS. Thus, our results may provide useful insights for the development of anti-sepsis agents via the sequence and structure information of the WIKE-14 peptide.

Immunomodulatory effects of Bacillus subtilis-fermented soybean extract in mice

ABSTRACT This study investigated the potential immunomodulatory effects of fermented soybean extract (FSE) in RAW264.7 cells and BALB/c mice. The expression of proinflammatory cytokines in RAW264.7 cells was measured after LPS stimulation in the presence or absence of FSE. Mice were orally administered 125, 250, and 500 mg/kg FSE for 21 days, and their immune response was analyzed. In RAW264.7 cells, FSE significantly reduced the expression of proinflammatory cytokines. In vivo experiments revealed significantly elevated splenocyte proliferation; neutrophil migration; phagocytic activities; expression of CD18+, CD4+, and CD8+ T cells; and expression of IL-2, IL-4, and IFN-γ. The major compounds in FSE were identified by GC–MS and by LC–MS/MS. Molecular docking analysis showed that daidzin, genistin and daidzein could potentially bind to the human high-affinity FcγRI–IgG complex to be involved in immunological responses. Taken together, FSE can be a candidate ingredient for health functional food with enhanced immunomodulating ability.

Designed symmetrical β-hairpin peptides for treating multidrug-resistant salmonella typhimurium infections

The rapid emergence and prevalence of multidrug-resistant salmonellosis lack effective therapies, which causes epidemic health problems and stimulates the development of antimicrobials with novel modes of action. In this research, 10 short symmetrical β-hairpin peptides are synthesized by combining the β-turn of Leucocin-A with recurring hydrophobic and cationic amino acid sequences. Those designed peptides exhibited potent antibacterial activities against drug-susceptible and drug-resistant Salmonella. One of the 10 peptides, WK2 ((WK)2CTKSGC(KW)2), displayed best cell selectivity towards Salmonella cells over macrophages and erythrocytes in a co-culture model. Fluorescent measurements and microscopic observations reflected that WK2 exerted its antimicrobial activity through a membrane-lytic mechanism. Moreover, the β-hairpin peptides can bind to endotoxin (LPS) and suppress the production of LPS-induced proinflammatory cytokines in RAW264.7 cells, indicating as a potent anti-inflammatory activity. The preliminary in vivo studies can also demonstrate that WK2 decreased loads of Salmonella in the liver and spleen, mitigated Salmonella-caused inflammation and maintained the integrity of intestinal mucosal surfaces. Ultimately, the results highlight that WK2 is a promising therapeutic agent to prevent multidrug-resistant S. Typhimurium infections in humans and animals.

Anti-Inflammatory Effects of Limosilactobacillus fermentum KGC1601 Isolated from Panax ginseng and Its Probiotic Characteristics.

We investigated the potential probiotic properties of Limosilactobacillus fermentum KGC1601 isolated from Panax ginseng. Ginseng cultivated in an experimental field of the Korea Ginseng Research Institute was fermented, followed by single colony selection from MRS agar. We performed 16s-rRNA sequencing and whole-genome analysis to identify L. fermentum and evaluate the biosafety parameters of this strain, respectively. We confirmed this strain was susceptible to six antibiotics, as proposed by the European Food Safety Authority, did not produce biogenic amines, and did not exhibit any hemolytic activity. Acid resistance and bile salt tolerance, which are essential properties of a probiotic agent, were investigated. Notably, distinguishing properties of this strain were that it exhibited excellent bile salt tolerance and anti-inflammatory effects. The excellent bile salt tolerance was confirmed by scanning electron microscopy. Through qRT-PCR and ELISA studies, it was revealed that L. fermentum KGC1601 pre-treatment up-regulates anti-inflammatory cytokines and down-regulates pro-inflammatory cytokines in RAW 264.7 cells. Consequently, we suggested that L. fermentum KGC1601 can be safely used as a potential anti-inflammatory functional probiotic agent.

Perillyl alcohol attenuates rheumatoid arthritis via regulating TLR4/NF-κB and Keap1/Nrf2 signaling pathways: A comprehensive study onin-vitro and in-vivo experimental models

BackgroundRheumatoid arthritis is a chronic and idiopathic autoimmune disorder. Perillyl alcohol (POH) is a monoterpene which can be extracted from widely available essential oils and is known for its strong anti-inflammatory and anti-oxidant properties. Hypothesis/PurposeRecent studies have been proven that inhibitors of farnesyltransferase enzyme showed significant anti-arthritic activity. POH is one such natural molecule having anti-inflammatory and anti-oxidant properties by inhibiting farnesyltransferase enzyme which further down regulates NF-κB and Nrf2 via Ras/Raf/MAPK pathway. Also, the effect of POH against rheumatoid arthritis is not known yet. Hence, the present research was intended to assess the anti-arthritic potential of POH in-vitro and in-vivo. MethodsThe in-vitro effects of POH on RAW 264.7 cells stimulated with LPS 1 µg/ml were investigated to its potential therapeutic effects. CFA 100 µl was intradermally administered to rats for the induction of arthritis. POH 100 mg/kg and 200 mg/kg administered topically from day 1 to day 28. Paw volumes measured, radiography analysis, anti-oxidant status, Gene expression studies, western blot analysis and histological analysis were performed to check the effects of POH. ResultsOur in-vitro findings suggested that POH inhibits inflammation by suppressing reactive oxygen species (ROS), NF-кB and Nrf2 signaling axis. Besides this, POH also rescinded the nitrate levels, pro-inflammatory cytokine levels like IL-1β, IL-6 and TNF-α also PGE2 and COX-2 levels induced by LPS in murine macrophages. Additionally, our in-vivo results revealed that POH conscientiously alleviated CFA induced inflammation by restoring arthritis index, body weight, nitrosative, lipid peroxidation assays. Macroscopically through measuring paw volumes and X-ray, it was evidenced that POH has decreased inflammation and bone erosion. Not only in-vitro but also in-vivo, POH has abridged cytokine levels IL-1β, IL-6, and TNF-α. Histopathological evaluation presented POH treatment alleviated joint inflammation, pannus formation, and bone erosion significantly. Moreover, POH suppressed the protein expression of NF-кB, COX-2, iNOS and improved Nrf2, and SOD2 levels in paw tissues estimated by western blotting. ConclusionPOH was effective in ameliorating LPS stimulation mediated oxidative stress and pro-inflammatory cytokines in RAW 264.7 cells in-vitro and FCA induced arthritis in rats in-vivo through its anti-inflammatory effects via regulating TLR4/NF-κB and Keap1/Nrf2 signaling pathways..

Elaeocarpus sylvestris extract (ESE) inhibits inflammatory mediators and increases the efficacy of sulfasalazine in rheumatoid arthritis

BackgroundInterest in plant-based therapeutic products for treatment of several autoimmune diseases, including rheumatoid arthritis (RA), is increasing as current RA medications and treatments are either very expensive or ineffective. PurposeWe investigated the anti-inflammatory activity of Elaeocarpus sylvestris extract (ESE) and its therapeutic potential in collagen-induced arthritis (CIA) in the DBA/1 J mouse model and an in vitro model. MethodsIn the in vitro assay, the effects of ESE and a combination of ESE + sulfasalazine (SLZN) on nitric oxide and proinflammatory cytokine expression induced by lipopolysaccharide in RAW264.7 cells were confirmed. Furthermore, enzyme-linked immunosorbent assays and real-time polymerase chain reactions were used to measure levels of RA-related inflammatory cytokines and biomarkers in serum and knee joints to evaluate the changes caused by co-administration of ESE and SLZN in our mouse CIA model. Micro-CT scans of knee joints were analyzed for all groups. ResultsESE and the combination of ESE + SLZN downregulated the expression of lipopolysaccharide-induced nitric oxide and proinflammatory cytokines in RAW264.7 cells. Also, combined ESE and SLZN treatment improved arthritis score, edema volume, and proinflammatory cytokine serum levels compared with the CIA group. In addition, chemokine expression was downregulated compared with the CIA group. ConclusionThese findings suggest that coadministration of ESE and SLZN can improve the efficacy of arthritis treatment with reduced side effects.

Structural Characterization of Sulfated Polysaccharide Isolated From Red Algae (Gelidium crinale) and Antioxidant and Anti-Inflammatory Effects in Macrophage Cells.

Gelidium crinale, the red algae belonging to Geliaceae Gelidium, is a traditional edible and industrial alga in China. A sulfated polysaccharide (GNP) is successfully separated from Gelidium crinale by acid extraction and two-step column chromatography. Chemical analysis showed that the molecular weight of GNP was 25.8 kDa and the monosaccharide composition had the highest galactose content and confirmed the presence and content (16.5%) of sulfate by Fourier transform infrared spectroscopy (FT-IR) spectrometry as well as barium chloride-gelatin methods. In addition, the effect of GNP on lipopolysaccharide (LPS)-induced oxidative stress and inflammation in macrophages was also evaluated. The research results showed that GNP had fairly strong scavenging activities on 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical, hydroxyl radical, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and had Fe2+-chelating ability in a dose-dependent manner. At the same time, it significantly inhibits the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) and the production of pro-inflammatory cytokines in RAW 264.7 cells induced by LPS through blocking the mitogen-activated protein kinase (MAPK)/nuclear factor kappa beta (NF-κB) signaling pathway. These results indicate that GNP may be a latent component anti-inflammation in pharmaceutical and functional food industries.

Aronia melanocarpa Extract Fermented by Lactobacillus plantarum EJ2014 Modulates Immune Response in Mice.

The present study aimed to assess the immunomodulatory effects of fermented Aronia melanocarpa extract (FAME) on RAW 264.7 cells and BALB/c mice. Aronia melanocarpa fruit was fermented with Lactobacillus plantarum EJ2014 by adding yeast extract and monosodium glutamate for 9 days at 30 °C to produce γ-aminobutyric acid (GABA). After fermentation, significant GABA production was noted, along with minerals, polyphenols, and flavonoids (p < 0.05). The polyphenol content was confirmed by liquid chromatography with tandem mass spectrometry (LC–MS/MS) analysis. RAW 264.7 cells were stimulated with lipopolysaccharide (LPS, 1 μg/mL) in the presence or absence of FAME, and proinflammatory cytokine contents were measured by qPCR. In the in vivo experiment, female BALB/c mice were administered 125, 250, and 500 mg/kg of FAME for 21 days. FAME treatment increased neutrophil migration and phagocytosis (p < 0.05). It also increased splenocyte proliferation, CD4+ and CD8+ T-cell expression, and lymphocyte proliferation. Furthermore, it increased IFN-γ, IL-2, and IL-4 cytokine levels in a dose-dependent manner (p < 0.05). However, it decreased TNF-α and IL-6 levels (p < 0.05). These results indicate that FAME fortified with GABA including bioactive compounds exerts anti-inflammatory effects by inhibiting proinflammatory cytokines in RAW 264.7 cells and modulates immune response in mice. Thus, FAME could be a potential therapeutic agent for inflammatory disorders.