Inducible Tumor Necrosis Factor-α Research Articles

Newly Synthesized Indolylacetic Derivatives Reduce Tumor Necrosis Factor-Mediated Neuroinflammation and Prolong Survival in Amyotrophic Lateral Sclerosis Mice.

The debilitating neurodegenerative disease known as amyotrophic lateral sclerosis (ALS) is characterized by the progressive loss of motor neurons (MNs) in the brain, spinal cord, and motor cortex. The ALS neuroinflammatory component is being characterized and includes the overexpression of mediators, such as inducible nitric oxide synthase (iNOS) and tumor necrosis factor-α (TNF-α). Currently, there are no effective treatments for ALS. Indeed, riluzole, an N-methyl-D-aspartate (NMDA) glutamate receptor blocker, and edaravone, a reactive oxygen species (ROS) scavenger, are currently the sole two medications approved for ALS treatment. However, their efficacy in extending life expectancy typically amounts to only a few months. In order to improve the medicaments for the treatment of neurodegenerative diseases, preferably ALS, novel substituted 2-methyl-3-indolylacetic derivatives (compounds II-IV) were developed by combining the essential parts of two small molecules, namely, the opioids containing a 4-piperidinyl ring with indomethacin, previously shown to be efficacious in different experimental models of neuroinflammation. The synthesized compounds were evaluated for their potential capability of slowing down neurodegeneration associated with ALS progression in preclinical models of the disease in vitro and in vivo. Notably, we produced data to demonstrate that the treatment with the newly synthesized compound III: (1) prevented the upregulation of TNF-α observed in BV-2 microglial cells exposed to the toxin lipopolysaccharides (LPS), (2) preserved SHSY-5Y cell survival exposed to β-N-methylamino-l-alanine (L-BMAA) neurotoxin, and (3) mitigated motor symptoms and improved survival rate of SOD1G93A ALS mice. In conclusion, the findings of the present work support the potential of the synthesized indolylacetic derivatives II-IV in ALS treatment. Indeed, in the attempt to realize an association between two active molecules, we assumed that the combination of the indispensable moieties of two small molecules (the opioids containing a 4-piperidinyl ring with the FANS indomethacin) might lead to new medicaments potentially useful for the treatment of amyotrophic lateral sclerosis.

Arctiin Protects Chondrocytes From Interleukin-1β-Induced Inflammation and Apoptosis by RNA Sequence In vivo and In vitro

Abstract Objective: Osteoarthritis (OA) is a progressive joint disease characterized by degeneration and destruction of articular cartilage. Arctiin (ARC) has been shown in many studies to have potential anti-inflammatory, anti-apoptotic, and antioxidant effects in various diseases. However, the mechanism by which ARC exerts its protective effects in OA is not fully understood. Here, we explore the mechanism by which ARC plays its protective role in OA. Materials and Methods: Mouse chondrocytes were isolated and characterized through toluidine blue staining and collagen II immunofluorescence labeling. A mouse-based experimental model was developed to induce chondrocyte inflammation through Interleukin-1β (IL-1β). Subsequently, ARC was administered in various doses to mitigate this inflammation. Techniques such as biochemical assays, Enzyme-linked immunosorbent assay, quantitative real-time polymerase chain reaction (qRT-PCR), Western blotting, and immunofluorescence labeling were employed to detect changes in nitric oxide (NO), lactate dehydrogenase (LDH), inflammatory markers, and components of the cartilage matrix in chondrocytes. RNA-sequencing (RNA-seq) was utilized to explore variations in gene expression among chondrocytes across different groups. The genes and signaling pathways that were identified underwent analysis through Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment studies. Validation of gene and protein expression was carried out using qRT-PCR, Western blotting, and cellular flow cytometry, based on the results from sequencing. Furthermore, Safranin-O fast green staining and immunohistochemistry staining were performed on slices of the mice knee joint to evaluate the OA Research Society International score, alterations in the cartilage matrix, and levels of apoptosis-related proteins at sites of knee cartilage damage in an arthritis model induced by monosodium iodoacetate (MIA) and physical activity. Results: It was found that ARC effectively inhibits the production of IL-1β-induced chondrocytes’ inducible NO synthase, cyclooxygenase-2, NO, LDH, IL-6, and tumor necrosis factor-α. ARC exhibited a dose-dependent effect on chondrocytes by reducing IL-1β-induced matrix metalloproteinase-3 (MMP-3) and a disintegrin and metalloproteinase with thrombospondin motifs-5 levels while increasing Aggrecan levels. RNA-seq and bioinformatics analysis revealed that ARC’s therapeutic effects involve apoptotic signaling pathways through the downregulation of Bcl-2-associated X protein (Bax) and caspase-3 expression and the upregulation of B-cell lymphoma-2 (Bcl-2) expression in IL-1β-induced chondrocytes. ARC significantly raised the levels of aggrecan and Bcl-2 and decreased the levels of MMP-3, Bax, and caspase-3 in an arthritis model induced by MIA and movement. Conclusions: Through RNA-seq, in vitro cell assays, and in vivo experiments, this research established the link between apoptosis and inflammation in the progression of OA and confirmed the protective effects of ARC on chondrocytes and its key targets. This highlights ARC’s therapeutic potential and its role in the development of treatments for OA.

유용미생물에 의한 저이용 산채류의 복합발효 중 생리활성 성분 및 항염증 변화

It was confirmed that complex fermentation (CF) was more efficient than single-strain fermentations in inducing changes in the contents of phenolic compounds of Maclura tricuspidate and Pyrus Montana Nakai. A mixture of Maclura tricuspidata, Pyrus montana Nakai, Platycodon grandiflorum and Codonopsis lanceolata were fermented in CF using Aspergillus shirousamii (koji), yeast, and lactic acid bacteria (LAB) for 24 days, and the pH, °Brix, total acidity, anti-oxidant activity, polyphenol content, nitric oxide (NO), and Western blotting of inducible nitric oxide synthase (iNOS), cyclo-oxygenase-2 (COX-2), and tumor necrosis factor-α (TNF-α) of the sample were determined. There was no significant change in pH and total acidity. °Brix significantly decreased from day 6 onwards. HPLC confirmed that the concentrations of chlorogenic acid, 4-hydrobenzoic acid, vanillic acid, and caffeic acid significantly increased from day 18 during the fermentation. Additionally, DPPH, ABTS radical scavenging activity, total phenol, and total flavonoid were confirmed to be increased until 18 days. NO was significantly inhibited from day 6, along with significant inhibition of iNOS, COX-2, and TNF-a. In conclusion, this study confirmed that CF of low-use (or underutilized) wild vegetables enhances phenolic compounds. It effectively suppresses NO, iNOS, COX-2, and TNF-α, markers of inflammation-related pathogenesis. Altogether, our results suggest that CF of the above plants has a potential anti-inflammatory effect.

The protective effect of Ergolide in osteoarthritis: In vitro and in vivo studies

Osteoarthritis (OA), a prevalent degenerative condition, occurs due to the deterioration of joint tissues and cells. Consequently, safeguarding chondrocytes against damage caused by inflammation is an area of future research emphasis. There is growing evidence that Ergolide (ERG) has multiple biological functions. Nevertheless, it is still uncertain whether it can hinder the advancement of OA. In this study, we investigate the ERG's potential to reduce inflammation and protect cartilage. ERG treatment in vitro effectively inhibited the excessive production of pro-inflammatory substances, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX2), and tumor necrosis factor-α (TNF-α), leading to their complete suppression. Furthermore, ERG suppressed the production of matrix-degrading enzymes (ADAMTS-5) and matrix metalloproteinase 13 (MMP13), consequently impeding the breakdown of extracellular matrix (ECM) and restraining the synthesis of collagenase II and Aggrecan. Through the P38/MAPK pathway, we discovered that ERG hinders the activation of NF-κB in chondrocytes induced by IL-1β. The protective effect of ERG was enhanced by the p38 MAPK inhibitor SB203580. In vivo, ERG further demonstrated protective effects on cartilage in animal models of DMM. In conclusion, the study has discovered that ERG exhibits innovative therapeutic potential in the context of OA.

Potential Benefits of Integrin αvβ3 Antagonists in a Mouse Model of Experimental Dry Eye.

The purpose of this study was to extensively evaluate the efficacy of integrin αvβ3 antagonists for the treatment of experimental dry eye (EDE). Vitronectin, an αvβ3 ligand, was used to induce tumor necrosis factor-α gene expression in human THP-1 macrophages. To induce EDE, C57BL/6 mice were housed in a low-humidity controlled environment chamber and injected subcutaneously with scopolamine for 7 days. Subsequently, αvβ3 antagonists, including RGDfD, c(RGDfD), c(RGDiD), c(RGDfK), ATN-161, SB273005, and cilengitide, were administered topically to EDE animals under controlled environment chamber conditions. Corneal epithelial damage in EDE was assessed by fluorescein staining. The density of conjunctival goblet cells and secretion of tears was measured by period acid-Schiff staining and phenol red-impregnated cotton threads, respectively. Inflammation markers, including tumor necrosis factor-α, interleukin (IL)-1β, IL-6, IL-17A, and metalloproteinase (MMP)-9, in the pooled cornea and conjunctiva tissues were examined by real-time polymerase chain reaction. The inhibitory effects of αvβ3 antagonists on the vitronectin-induced tumor necrosis factor-α gene expression and integrin-mediated inflammatory signaling were validated in THP-1 macrophages. αvβ3 antagonists ameliorated the impairment of the corneal epithelial barrier with varying therapeutic efficacies, compared with vehicle-treated mice. c(RGDfD) and c(RGDiD) significantly protected against goblet cell loss, tear reduction, and proinflammatory gene expression in EDE. Topical applications of αvβ3 antagonists yield therapeutic benefits in EDE by promoting corneal epithelial defect healing and reducing inflammation. Antagonistic targeting αvβ3 may be a novel promising strategy to treat patients with dry eye disease.

In Vitro and In Vivo Anti-Inflammatory Potential of Cannabichromene Isolated from Hemp

Cannabichromene (CBC), a non-psychoactive cannabinoid found in Cannabis sativa, has recently been shown to possess several medicinal properties. However, how CBC produces anti-inflammatory effects and the mechanisms of this remain poorly studied. Therefore, we extracted and purified the CBC from the Cannabis sativa cv. pink pepper (hemp cultivar). The efficacy of CBC in reducing inflammation in RAW 264.7 macrophages and a λ-carrageenan-induced mouse model was then evaluated. CBC had no cytotoxicity up to a concentration of 20 μM and inhibited nitric oxide production by approximately 50% at a concentration of 20 μM. In addition, CBC treatment significantly inhibited causes of inflammation such as inducible nitric oxide synthase (iNOS), interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α) at both the mRNA and protein levels. Moreover, CBC suppressed LPS-stimulated inflammation in RAW 264.7 cells by downregulating the nuclear factor kappa B (NF-kB) and mitogen-activated protein kinase pathways (MAPK). Furthermore, our in vivo experiments confirmed that the λ-carrageenan-induced increase in the levels of the cytokines iNOS, IL-1β, and IL-6 was abrogated following treatment with CBC. Therefore, CBC has potential anti-inflammatory effects and may be useful for preventing or treating inflammation.

Anti-Neuroinflammatory Effect of the Ethanolic Extract of Black Ginseng through TLR4-MyD88-Regulated Inhibition of NF-κB and MAPK Signaling Pathways in LPS-Induced BV2 Microglial Cells.

Korean ginseng (Panax ginseng) contains various ginsenosides as active ingredients, and they show diverse biological activities. Black ginseng is manufactured by repeated steaming and drying of white ginseng, which alters the polarity of ginsenosides and improves biological activities. The aim of the present investigation was to examine the anti-neuroinflammatory effects of the ethanolic extract of black ginseng (BGE) in lipopolysaccharide (LPS)-induced BV2 microglial cells. Pre-treatment with BGE inhibited the overproduction of pro-inflammatory mediators including nitric oxide (NO), prostaglandin E2 (PGE2), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in LPS-induced BV2 cells. In addition, BGE reduced the activation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), p38 mitogen-activated protein kinase (MAPK), and c-jun N-terminal kinase (JNK) MAPK signaling pathways induced by LPS. These anti-neuroinflammatory effects were mediated through the negative regulation of the toll-like receptor 4 (TLR4)/myeloid differentiation primary response 88 (MyD88) signaling pathway. Among the four ginsenosides contained in BGE, ginsenosides Rd and Rg3 inhibited the production of inflammatory mediators. Taken together, this investigation suggests that BGE represents potential anti-neuroinflammatory candidates for the prevention and treatment of neurodegenerative diseases.

Magnesium sulfate enhances lipopolysaccharide tolerance

Introduction Lipopolysaccharide (LPS) tolerance is the downregulation of LPS signaling after pre-exposure to LPS, and it provides protection against hyperactive inflammation. Cytokine production decreases during LPS tolerance, and the phenotype of LPS-tolerant monocytes shifts toward M2 (anti-inflammatory) type. Magnesium sulfate (MgSO4) is a widely used anti-inflammatory agent. Although MgSO4 inhibits LPS signaling, the effect of MgSO4 on LPS tolerance is unknown. In the present study, we investigated the in vitro effects of MgSO4 on LPS tolerance. Methods To induce LPS tolerance, THP-1 cells were stimulated with LPS (200 ng/mL, 2 h) after pre-exposure to LPS (200 ng/mL, 24 h) with or without pre-treatment of MgSO4 (20 mM, 24 h). Results Our results revealed that MgSO4 enhanced LPS tolerance by downregulating nuclear factor-κB (NF-κB)-induced tumor necrosis factor-α or interleukin-6, and upregulating cluster of differentiation 163 (a M2-associated marker). Furthermore, the LPS-triggered upregulation of phosphoinositide 3-kinase (PI3K) was significantly increased during LPS tolerance. MgSO4 activated PI3K, but inhibited NF-κB in LPS-stimulated cells. Notably, MgSO4 mitigated the signaling of both PI3K and NF-κB in LPS-tolerant cells, suggesting the effect of MgSO4 on LPS tolerance relies on the modulation of the crosstalk between PI3K and NF-κB. Conclusion MgSO4 enhanced LSP tolerance, thus providing evidence for a novel underlying mechanism of the anti-inflammatory effects of MgSO4.

The protective effects of a novel AT2 receptor agonist, β-Pro7Ang III in ischemia-reperfusion kidney injury

Background and purposeThis study investigated the reno-protective effects of a highly selective AT2R agonist peptide, β-Pro7Ang III in a mouse model of acute kidney injury (AKI). MethodsC57BL/6 J mice underwent either sham surgery or unilateral kidney ischemia-reperfusion injury (IRI) for 40 min. IRI mice were treated with either β-Pro7Ang III or perindopril and at 7 days post-surgery the kidneys analysed for histopathology and the development of fibrosis and matrix metalloproteinase (MMP)-2 and -9 activity. The association of the therapeutic effects of β-Pro7Ang III with macrophage number and phenotype was determined in vivo and in vitro. Key resultsDecreased kidney tubular injury, interstitial matrix expansion and reduced interstitial immune cell infiltration in IRI mice receiving β-Pro7Ang III treatment was observed at day 7, compared to IRI mice without treatment. This correlated to reduced collagen accumulation and MMP-2 activity in IRI mice following β-Pro7Ang III treatment. FACS analysis showed a reduced number and proportion of CD45+CD11b+F4/80+ macrophages in IRI kidneys in response to β-Pro7Ang III, correlating with a significant increase in M2 macrophage markers and decreased M1 markers at day 3 and 7 post-IR injury, respectively. In vitro analysis of cultured THP-1 cells showed that β-Pro7Ang III attenuated lipopolysaccharide (LPS)-induced tumour necrosis factor-α (TNF-α) and interleukin (IL)− 6 production but increased IL-10 secretion, compared to LPS alone. ConclusionAdministration of β-Pro7Ang III via mini-pump improved kidney structure and reduced interstitial collagen accumulation, in parallel with an alteration of macrophage phenotype and anti-inflammatory cytokine release, therefore mitigating the downstream progression of ischemic AKI.

In Vitro Anti-Inflammatory and Antioxidant Activities of pH-Responsive Resveratrol-Urocanic Acid Nano-Assemblies.

Inflammatory environments provide vital biochemical stimuli (i.e., oxidative stress, pH, and enzymes) for triggered drug delivery in a controlled manner. Inflammation alters the local pH within the affected tissues. As a result, pH-sensitive nanomaterials can be used to effectively target drugs to the site of inflammation. Herein, we designed pH-sensitive nanoparticles in which resveratrol (an anti-inflammatory and antioxidant compound (RES)) and urocanic acid (UA) were complexed with a pH-sensitive moiety using an emulsion method. These RES-UA NPs were characterized by transmission electron microscopy, dynamic light scattering, zeta potential, and FT-IR spectroscopy. The anti-inflammatory and antioxidant activities of the RES-UA NPs were assessed in RAW 264.7 macrophages. The NPs were circular in shape and ranged in size from 106 to 180 nm. The RES-UA NPs suppressed the mRNA expression of the pro-inflammatory molecules inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages in a concentration-dependent manner. Incubation of LPS-stimulated macrophages with RES-UA NPs reduced the generation of reactive oxygen species (ROS) in a concentration-dependent manner. These results suggest that pH-responsive RES-UA NPs can be used to decrease ROS generation and inflammation.

Docosahexaenoic acid inhibits TNF-α-induced osteoclast formation and orthodontic tooth movement through GPR120.

Docosahexaenoic acid (DHA) is an omega-3 fatty acid that has a range of positive impacts on human health, including anti-inflammatory effects and inhibition of osteoclast formation via G-protein-coupled receptor 120 (GPR120). Orthodontic force was reported to induce tumor necrosis factor-α (TNF-α) expression, which activates osteoclast differentiation during orthodontic tooth movement (OTM). The aim of this study was to investigate the influence of DHA on TNF-α-induced osteoclast formation and OTM in vivo. We examined osteoclast formation and bone resorption within the calvaria of both wild-type (WT) and GPR120-deficient (GPR120-KO) mice injected with phosphate-buffered saline (PBS), TNF-α, TNF-α and DHA, or DHA. DHA inhibited TNF-α-induced osteoclast formation and bone resorption in WT mice but had no effect in GPR120-KO mice. OTM experiments were performed in mouse strains with or without regular injection of DHA, and the effects of DHA on osteoclast formation in the alveolar bones during OTM were examined. DHA also suppressed OTM in WT but not GPR120-KO mice. Our data showed that DHA suppresses TNF-α-induced osteoclastogenesis and bone resorption via GPR120. TNF-α has considerable significance in OTM, and therefore, DHA may also inhibit TNF-α-induced osteoclast formation and bone resorption in OTM.

Yomogin, Isolated from Artemisia iwayomogi, Inhibits Neuroinflammation Stimulated by Lipopolysaccharide via Regulating MAPK Pathway.

Neuroinflammation causes various neurological disorders, including depression and neurodegenerative diseases. Therefore, regulation of neuroinflammation is a promising therapeutic strategy for inflammation-related neurological disorders. This study aimed to investigate whether yomogin, isolated from Artemisia iwayomogi, has anti-neuroinflammatory effects. First, we evaluated the effects of yomogin by assessing pro-inflammatory mediators and cytokines in lipopolysaccharide (LPS)-stimulated BV2 microglial cells. The results showed that yomogin inhibited the increase in neuroinflammatory factors, including nitric oxide, inducible nitric oxide synthase, cyclooxygenase-2, interleukin-6, and tumor necrosis factor-α, and suppressed phosphorylation of c-Jun N-terminal kinase, extracellular signal-regulated kinase and p38, which participate in the mitogen-activated protein kinase (MAPK) pathway. To confirm these effects in vivo, we measured the activation of astrocyte and microglia in LPS-injected mouse brains. Results showed that yomogin treatment decreased astrocyte and microglia activations. Collectively, these results suggest that yomogin suppresses neuroinflammation by regulating the MAPK pathway and it could be a potential candidate for inflammation-mediated neurological diseases.

The relationship between leukocyte level and hypertension in elderly patients with hyperuricemia

To evaluate the change of leukocyte level caused by hyperuricemia, and to explore the relationship between leukocyte level and hypertension in elderly patients with hyperuricemia. A cross-sectional study of serum uric acid (UA) level was conducted in 1352 elderly people over 65 years old. The samples were divided into 3 categories according to the tertiles of leukocyte: Tertile 1, leukocyte ≤ 5.2 × 109/L; Tertile 2, leukocyte = 5.3-6.3 × 109/L; Tertile 3, leukocyte ≥ 6.4 × 109/L. Multiple logistic regression models were used for modeling relationships between leukocyte, hyperuricemia and hypertension. Human vascular endothelial cells were treated by different concentrations of UA. The levels of interleukin-1 beta, tumor necrosis factor-α, endothelial nitric oxide synthase, inducible nitric oxide synthase and reactive oxygen species were measured by Western Blot or fluorescence microscope. The levels of leukocyte were higher in elderly patients with hyperuricemia than without hyperuricemia. Hyperuricemia was an independent risk factor of leukocyte in Tertile 3 (odds ratio [OR] = 1.657, 95% confidence interval [CI]: 1.180-2.328). The prevalences of hypertension were higher in elderly patients with hyperuricemia than without hyperuricemia (77.0% vs 63.5%). In the Model 1, hyperuricemia was an independent risk factor of hypertension (OR = 1.536, 95% CI: 1.026-2.302). Leukocyte in Tertile 3 was an independent risk factor of hypertension (OR = 1.333, 95% CI: 1.031-1.724). Expression levels of interleukin-1 beta, inducible nitric oxide synthase and tumor necrosis factor-α were obviously higher in the UA group than the control group, along with the productions of reactive oxygen species. But the expression level of endothelial nitric oxide synthase was obviously lower in the UA group. Hyperuricemia was associated with an increased risk for hypertension. The chronic inflammation caused by hyperuricemia maybe one of important pathogenesis of incident hypertension in patients with hyperuricemia.

Levosimendan Increases Survival in a D-Galactosamine and Lipopolysaccharide Rat Model.

Levosimendan, a calcium sensitizer, has an organ protective profile through the inhibition of inflammatory mediators and cytokines in critical conditions, such as heart failure, ischemia-reperfusion injury, and sepsis. The survival effect of levosimendan for acute liver failure has not been examined yet. Male Sprague-Dawley rats were examined in the D-galactosamine hydrochloride and lipopolysaccharide (GalN/LPS) model. Levosimendan was injected intraperitoneally before GalN/LPS treatment. Survival was monitored for 7 days. For biochemical analyses, liver and blood samples were collected from the rats at 1 and 8 h after GaIN/LPS treatment. The pretreatment of levosimendan at 4 mg/kg significantly increased survival in GalN/LPS rats. In the liver specimen, levosimendan significantly inhibited the activation of nuclear factor-κB (NF-κB) at 1 h, and significantly decreased the mRNA expression of inflammatory mediators, including inducible nitric oxide synthase and tumor necrosis factor-α (TNF-α), at 8 h. In serum, levosimendan decreased the levels of nitrite, a metabolite of nitric oxide, and TNF-α protein, as well as aspartate aminotransferase and alanine aminotransferase. These results indicated that Levosimendan ameliorated liver dysfunction and survival in acute liver failure model rats through the suppression of NF-κB activation.

Steroidal Saponins from the Rhizomes of Smilax china and Their Inhibitory Effects on Lipopolysaccharide-Induced Proinflammatory Cytokines Expression.

Four new furostanol saponins (1: - 4: ) and a new pregane-type saponin (5: ) along with six known steroidal saponins (6: - 11: ) were isolated from the rhizomes of Smilax china. The structures of 1: - 5: were elucidated by extensive analysis of NMR and HR-ESI-MS data in addition to enzymatic hydrolysis and other chemical methods. Compounds 1, 4: , and 11: showed inhibitory activity against the expression of proinflammatory mediators, inducible nitric oxide synthase, interleukin-1β, interleukin-6, and tumor necrosis factor-α in lipopolysaccharide-induced RAW264.7 cells. Compound 1: , at a concentration of 20 µM, decreased the production of inducible nitric oxide synthase, interleukin-1β, interleukin-6, and tumor necrosis factor-α by 36, 62, 72, and 67%, respectively, which is comparable to that of the positive control dexamethasone.

Immunotoxic potential of nanoparticles of cerium oxide and gadolinium oxide in human monocyte (THP-1) cells

ObjectivesNPs of cerium oxide (CeO2) and gadolinium oxide (Gd2O3; designated as GO for simplicity) belong to the lanthanide group of elements. Due to the potential applications of GO NPs in MRI or CeO2 NPs as novel antioxidant, attention about their impact on the immune-toxicity is needed. MethodsPotential immune-toxicity response due to interactions of GO NPs (54.8 ± 29 nm in length and 13.7 ± 6 nm in diameter with a fiber-like shape) and CeO2 NPs (40 ± 17 nm with a cuboidal shape) was examined in human monocytic THP-1 cells exposed for 24 h, 48 h and 72 h. A battery of parameters intended to measure oxidative stress and mechanism of cell death, if occurring, in THP-1 cells exposed to NPs for the periods as mentioned above was also examined. ResultsNPs of GO induced higher toxicity and oxidative stress than it was by NPs of CeO2. Interestingly, CeO2 NPs behaved as antioxidant when exposed for 24 h at concentrations of 100- and 200 µg/mL. This trend, however, reversed at 72 h of exposure that resulted in induction of significant cytotoxicity, reactive oxygen species (ROS) and exhaustion of cellular antioxidant glutathione (GSH). Cuboid CeO2 NPs with low potential to induce toxicity and oxidative stress caused a rise in Bax/Bcl2 ratio and caspase 3 apoptotic markers in THP-1 cells. Lack of these apoptotic markers in more toxic fiber-like GO NPs treated THP-1 cells indicate an apoptosis-independent mechanism of cell death by GO NPs. Although, CeO2 NPs caused induction of interleukin-1β (IL-1β) after 72 of exposure, CeO2 NPs did not induce tumor necrosis factor-α (TNF-α) at any duration of exposure. ConclusionDespite belonging to same rare-earth metal groups in lanthanide series, NPs of GO and CeO2 induce different bio-responses in human immune THP-1 cells. GO NPs were highly inflammatory than the NPs of CeO2. To the best of our knowledge, this is the first report on comparative analysis carried out for these NP’s potential cytotoxicity, oxidative stress and inflammation in THP-1 cells.

Qingchang Wenzhong Decoction Alleviates DSS-Induced Inflammatory Bowel Disease by Inhibiting M1 Macrophage Polarization In Vitro and In Vivo.

An imbalance of macrophage M1/M2 polarization significantly influences the pathogenesis of inflammatory bowel disease. Qingchang Wenzhong decoction (QCWZD) has a proven therapeutic effect on patients with inflammatory bowel disease (IBD) and can significantly inhibit the inflammatory response in mice with colitis. However, its effect on macrophages during IBD treatment remains nebulous. Aim of the Study. Explore the mechanism underlying QCWZD effects in a dextran sulfate sodium (DSS)-induced colitis mouse model in vivo and RAW264.7 cell in vitro by observing macrophage polarization dynamics. The main active components of QCWZD were determined using high-performance liquid chromatography. Surface marker expression on M1-type macrophages was analyzed using flow cytometry and immunofluorescence. The effect on inducible nitric oxide synthase (iNOS), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) released by M1 type macrophages was determined using ELSA and RT-PCR. The expression of key proteins in the JAK2/STAT3 signaling pathway was analyzed using western blotting. QCWZD cytotoxicity in macrophages was measured using CCK8 and Annexin V-FITC/PI assays. The main active components of QCWZD were berberine chloride, coptisine chloride, epiberberine chloride, gallic acid, ginsenoside Rg1, ginsenoside Rb1, indigo, indirubin, notoginsenoside R1, palmatine chloride, and 6-curcumin. QCWZD markedly alleviated DSS-induced colitis in mice, as revealed by the rescued weight loss and disease activity index, attenuated the colonic shortening and mucosal injury associated with the inhibition of M1 macrophage polarization and expression of related cytokines, such as IL-6 and TNF-α, in vivo and in vitro. Furthermore, QCWZD decreased the iNOS, JAK2, and STAT3 levels in vivo and in vitro, regulating the JAK2/STAT3 signaling pathway. QCWZD administration improves intestinal inflammation by inhibiting M1 macrophage polarization. The JAK2/STAT3 signaling pathway may mediate the effects of QCWZD on M1 macrophage polarization in colitis treatment. This study presents a novel macrophage-mediated therapeutic strategy for the treatment of IBD.

Expression of concern on: Mitogen-activated protein kinases regulate Mycobacterium avium-induced tumor necrosis factor-α release from macrophages.

Expression of concern on: Mitogen-activated protein kinases regulate Mycobacterium avium-induced tumor necrosis factor-α release from macrophages Get access Pathogens and Disease, Volume 80, Issue 1, 2022, ftac017, https://doi.org/10.1093/femspd/ftac017 Published: 22 June 2022

Amelioration of TPA-induced skin inflammation by the leaf extract of Vernonia amygdalina involves ERK/STAT3 (Ser727) signaling inhibition

BackgroundUncontrolled inflammation causes health problems. Extracellular signal-regulated kinase (ERK) phosphorylates signal transducer and activator of transcription 3 (STAT3) at Ser727, resulting in inflammation. The leaf of Vernonia amygdalina (VA) is a medicinal herb for managing inflammation-associated diseases. Oral administration or topical application of VA leaf extract exerts anti-inflammatory effects in rat models. However, the anti-inflammatory mechanisms of the herb are not fully understood. PurposeIn this study, we aimed to investigate the involvement of ERK/STAT3 (Ser727) signaling in the anti-inflammatory effects of an ethanolic extract of VA leaves. Study design and methodsExtracts of VA leaves were prepared with different concentrations of ethanol. A LPS-stimulated RAW264.7 cell model was used for in vitro assays, and a TPA (12-O-tetradecanoylphorbol-13-acetate)-induced ear edema mouse model was employed for in vivo assays. The 95% ethanol extract of VA leaves (VAE) exerted the strongest inhibitory effect on nitric oxide (NO) production in LPS-stimulated macrophages; thus it was selected for use in this study. Hematoxylin and eosin (H&E) staining was used to examine pathological conditions of mouse ear tissues. Griess reagent was employed to examine NO generation in cell cultures. Immunoblotting and ELISA were used to examine protein levels, and RT-qPCR was employed to examine mRNA levels. ResultsTopical application of VAE ameliorated mouse ear edema induced by TPA. VAE suppressed the phosphorylation of ERK (Thr202/Tyr204) and STAT3 (Ser727); and decreased protein levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin (IL)-6, IL-1β and tumor necrosis factor-α (TNF-α) in the mouse ear tissues and in LPS-stimulated RAW 264.7 cells. VAE also inhibited NO production, and lowered mRNA levels of IL-6, IL-1β and TNF-α in the macrophages. ConclusionsVAE ameliorates TPA-induced mouse ear edema. Suppression of ERK/STAT3 (Ser727) signaling is involved in VAE's anti-inflammatory effects. These novel data provide further pharmacological justifications for the medicinal use of VA in treating inflammation-associated diseases, and lay the groundwork for developing VAE into a new anti-inflammatory agent.

The ameliorative effect of Acetylsalicylic acid plus Ascorbic acid against renal injury in Corn Syrup-fed rats

Dietary consumption of commercially prepared (often through processing) corn syrup can activate reactive oxygen species and the inflammatory pathways observed in kidney damage in humans and experimental animals. This study was designed to assess the effects of antioxidant properties of acetylsalicylic (ASA) and ascorbic acid (AA) on corn syrup (CS) induced renal injury. Forty male Sprague–Dawley rats (14–16 weeks) were randomly classified into 5 groups, each containing 8 rats: control group (C), CS group, CS+ASA group (ASA-10 mg/kg/day/po), CS+AA group (AA-200 mg/kg/day/po) and CS+ASA+AA group (combination treatment with ASA and AA doses). At the end of the experiment, blood and tissue samples were collected for biochemical, histopathological and immunohistochemical examinations. Malondialdehyde (p<0.001) levels were increased in the CS group compared with control group and were decreased in the ASA, AA and ASA+AA treated groups (p≤0.001 for all) compared with CS group. Catalase activities were decreased in the CS group (p≤0.001) compared with the control group (p<0.001) and significantly increased in AS+AA-treated group (p<0.001) compared with the CS group. Significant histopathological changes including tubular vacuoler degeneration, tubular dilatation, cortical and medullar haemorrhage, mononuclear cell infiltration, and increased inducible nitric oxide synthase and tumor necrosis factor-α staining were observed in the CS group. Combination theraphy reversed all these changes. The administration of ASA and AA for the treatment of kidney damage due to corn syrup consumption provides an ideal target for improving oxidative stress and potential therapeutic treatments.