Pro-inflammatory Cytokines TNF-α Research Articles

Intermittent Fasting Reduces Intestinal Inflammation in Dextran Sulfate Sodium-Induced Colitis of Mice.

Inflammatory bowel disease (IBD), comprising ulcerative colitis (UC) and Crohn's disease (CD), is a chronic condition impacting both the gastrointestinal tract and the immune system. Intestinal inflammation and epithelial injury are the pathological features of IBD. Recent studies have reported that some strategies of dietary restriction (DR) can regulate immune system, correct the immune disorders, and improve some immune-associated diseases such as IBD. However, as a form of DR, the effect of intermittent fasting (IF) on the IBD remains unknown. In this study, we investigated the therapeutic efficacy of two cycles of IF on the IBD mouse model induced by dextran sulfate sodium (DSS). It was found that two cycles of IF significantly decreased the score of the disease activity index (DAI) and alleviated the IBD-related symptoms. In addition, IF reversed the shortening of colon length mediated by DSS, significantly increased the number of colonic crypts, and decreased the colonic histological score. Furthermore, the proportion of CD4+ T cells in both the spleen and mesenteric lymph node was reduced by IF treatment. The expression of serum pro-inflammatory cytokines IL-1β, TNF-α, and IL-6 was restrained by IF intervention. Moreover, IF administration significantly reduced the number of leukocytes and macrophages infiltrating around the crypt base in the colon. In conclusion, these results demonstrated that IF administration can alleviate the symptoms and pathology of IBD in the DSS-induced IBD mouse model by reducing the intestinal inflammation.

Delayed Immune Response Upon Injury in Diabetic Wounds Impedes Healing.

Chronic wounds are a severe complication of diabetes. Dysregulated inflammatory signalling is thought to underly the poor healing outcomes. Yet, there is little information available on the acute response following injury and its impact on healing. Using a murine full thickness excisional wound model, the current study therefore assessed the expression of pro-inflammatory and pro-resolving lipid mediators during the early stages post injury in acute and diabetic wounds and compared the timeframe for transitioning through the phases of healing. Tissue eicosanoid (LTB4, PGE2, TxA2, MaR1, RvE1, RvD1, PD) and MMP-9 levels were assessed at 6 h post wounding using ELISAs. Wound closure, healing dynamics (histology), cellular infiltration and MPO, TNF-α expression (IHC) were assessed at 6 h, day2, day7 post wounding. Eicosanoid expression did not differ between groups (LTB4 24-125 pg/mL, PGE2 63-177 pg/mL, TxA2 529-1184 pg/mL, MaR1 365-2052 pg/mL, RvE1 43-1157 pg/mL, RvD1 1.5-69 pg/mL, PD1 11.5-4.9 ng/mL). An inverse relationship (p < 0.05) between MMP-9 and eicosanoids were however only evident in acute and not in diabetic wounds. Diminished cellular infiltration (x5 fold) (p < 0.05) in diabetic wounds coincided with a significant delay in the expression of TNF-α (pro-inflammatory cytokine) and MPO (neutrophil marker). A significant difference in the expression of TNF-α (C 1.8 ± 0.6; DM 0.7 ± 0.1 MFI) and MPO (C 4.9 ± 1.9; DM 0.9 ± 0.4 MFI) (p < 0.05) was observed as early as 6 h post wounding, with histology parameters supporting the notion that the onset of the acute inflammatory response is delayed in diabetic wounds. These observations imply that the immune cells are unresponsive to the initial eicosanoid expression in the diabetic wound tissue.

Effect of 3D-Printed Polycaprolactone Scaffold With Powdery/Smooth Micromorphology on Local Immune Environments.

Selective laser sintering (SLS) has become a viable approach for producing biodegradable medical implants in various clinical applications. The resulting scaffolds typically exhibit a powdery microstructure, which may potentially impact the behavior of immune cells and immune responses in surrounding tissues. However, limited research has been conducted to understand the effect of surface morphology in SLS-fabricated scaffolds on local immune environments. This study aims to compare the effect of SLS-fabricated polycaprolactone (PCL) scaffolds with powdery and smooth surface morphologies on immune-related biological responses. Compared with those on the powdery micromorphology, RAW264.7 macrophages displayed greater dispersion and adopted a spread and elongated morphology on the scaffolds with smooth surface. The expression levels of arginase-1 and CD206 were found to be upregulated in macrophages adhering to the PCL scaffolds with smooth surface, accompanied by an augmented secretion of anti-inflammatory cytokines TGF-β and IL-10. Conversely, there was a decrease in the secretion of pro-inflammatory cytokines TNF-α and IL-12. When implanted invivo, the SLS-derived scaffolds were completely covered by host tissues, Withing increased collagen deposition, indicating good histocompatibility. At 1-week post-implantation, there was a significantly higher presence of M2-type macrophages surrounding the scaffold compared to M1 macrophages in both groups. By 3 weeks post-implantation, the overall level of macrophages had decreased in both groups. However, a significant higher level of M1 macrophages were observed in the powdery scaffold group. At the same time, the number of neutrophils around the powder scaffold increased significantly, demonstrating long-term local inflammatory responses. The results suggested that post-treated scaffolds with smooth surfaces can effectively reduce local inflammation, making them more suitable for clinical implantation.

Adjunctive Treatment Effect of Non-Thermal Atmospheric Pressure Plasma in Periodontitis-Induced Rats

Background/Objectives: As non-thermal atmospheric pressure plasma (NTP) is known to have advantages in application in the medical field, we consider its applicability to periodontitis, a representative chronic inflammatory disease. The purpose of this study was to evaluate the effect of NTP in inhibiting the progression of periodontitis in a rat model when additionally used in scaling and root planing (SRP). Methods: To induce experimental periodontitis in 20 rats, ligatures were placed in the maxillary second molar and lipopolysaccharide from Porphyromonas gingivalis was injected around the teeth. Then, NTP treatment was performed for 2 or 5 min, together with scaling and root planing (SRP). To evaluate alveolar bone loss, micro-computed tomography (micro-CT) analysis and hematoxylin–eosin (H-E) staining were performed. Tartrate-resistant acid phosphatase (TRAP) analysis was performed to compare the number of osteoclasts, while immunohistochemistry (IHC) analysis was performed to determine the expression levels of receptor activator of nuclear factor-𝜅B ligand (RANKL) and osteoprotegerin (OPG). Enzyme-linked immunosorbent assay (ELISA) analysis was performed for the detection of cytokines (TNF-α, IL-1β, and IL-10) in tissues and sera. Results: When SRP was combined with NTP, alveolar bone loss was decreased, the number of osteoclasts and RANKL expression were decreased, OPG expression was increased, and pro-inflammatory cytokine (TNF-α and IL-1β) levels were significantly decreased. Compared with the NTP treatment for 2 min, when treated for 5 min, less alveolar bone loss, fewer osteoclasts, a lower RANKL expression level, and a higher OPG expression level were observed. Conclusions: This study evaluated the adjunctive treatment effect of NTP in periodontitis-induced rats. Based on the results of this study, we suggest that supplemental NTP treatment may be a good option for non-surgical periodontal treatment; however, further studies are needed to elucidate the mechanism through which NTP suppresses periodontal inflammation.

Dietary supplementation of mulberry leaf oligosaccharides improves the growth, glucose and lipid metabolism, immunity, and virus resistance in largemouth bass (Micropterus salmoides)

This study investigated the effects of dietary supplementation of mulberry leaf oligosaccharides (MLO) on the growth performance, serum biochemistry, glucose and lipid metabolism, antioxidant activity, liver health, and virus resistance in largemouth bass (Micropterus salmoides). The fish were fed with CK (basal diet), MLOL (basal diet supplemented with 0.5%MLO), and MLOH (basal diet supplemented with 1.0% MLO) for 80 days, and then subjected to a 21-day viral challenge experiment. The results showed that MLO supplementation had no adverse effect on the weight gain rate, specific growth rate, feed intake, and condition factor (P &amp;gt; 0.05), but significantly decreased the feed conversion rate and viscerosomatic index (P&amp;lt; 0.05). Moreover, the MLOL and MLOH group had significantly lower contents of triglyceride, blood glucose, and malondialdehyde and activities of serum alanine aminotransferase and aspartate aminotransferase, while significantly higher levels of serum and liver total superoxide dismutase and lower levels of glutathione than the CK group (P&amp;lt; 0.05). MLO supplementation significantly up-regulated the relative expression of glycolytic genes gk and pfk and lipid catabolism genes ppar-α and cpt-1, while obviously down-regulated that of acc, fas, and dgat related to fatty acid synthesis in the liver tissue (P&amp;lt; 0.05). In terms of liver health, MLO supplementation significantly up-regulated the relative expression of anti-inflammatory cytokines il-10 and tgf-β, while decreased that of pro-inflammatory cytokines nf-κb, il-8, and tnf-α in the liver tissue (P&amp;lt; 0.05). The viral challenge test showed that MLO supplementation significantly improved the survival rate of M. salmoides after largemouth bass ranavirus (LMBV) infection. Dietary MLO supplementation promoted liver glucose and lipid metabolism, and improved the immunity and resistance of M. salmoides to LMBV by regulating the PPAR signaling way and inhibiting the NF-kB signaling pathway. The appropriate addition amount of MLO to the diet was determined to be 1.0%.

Porphyromonas gingivalis affects neutrophil pro-inflammatory activities

Porphyromonas gingivalis is the primary pathogen responsible for the development of periodontal inflammatory disease. Although gingipains are the major virulence factor of the pathogen, their role in impairing apoptosis and immune cell function is not fully understood. To investigate the impact of gingipains on neutrophil viability and function, we conducted studies using murine HoxB8 neutrophils and primary human neutrophils infected with wild-type strains of Porphyromonas gingivalis (W83 and ATCC 33277), or a gingipains-null mutant with deleted gingipains encoding genes, or wild-type bacteria preincubated with specific gingipain inhibitors. Flow cytometry revealed that wild-type Porphyromonas gingivalis had a marked effect on neutrophil viability regulated by anti-apoptotic proteins belonging to the Bcl-2 family; however, these effects were independent of gingipain expression or activity. Importantly, experiments using primary human neutrophils and macrophages revealed that gingipains play a significant role in the disruption of immune cell functions via the induction of reactive oxygen species and inactivation of neutrophil elastase activity. Additionally, although gingipains played a role in modulating the IL-8-dependent inflammatory response of human neutrophils, they did not affect the expression levels of pro-inflammatory cytokines TNF-α and IL-6.

Kurarinone Mitigates LPS-Induced Inflammatory Osteolysis by Inhibiting Osteoclastogenesis Through the Reduction of ROS Levels and Suppression of the PI3K/AKT Signaling Pathway.

Inflammatory bone resorption represents a pathological condition marked by an increase in bone loss, commonly associated with chronic inflammatory conditions such as rheumatoid arthritis and periodontitis. Current therapies primarilyfocuson anti-inflammatory drugs and bisphosphonates; however, these treatments are limited due to side effects, inadequate efficacy, and unpredictable long-term complications. Kurarinone (KR), a bioactive compound isolated from the traditional Chinese herb Sophora flavescens, exhibits a range of biological activities, including anti-inflammatory, anticancer, and cardiovascular protective effects. To address the limitations of existing therapies and enhance drug utilization, this study explores the potential of KR as a therapeutic agent for inflammatory bone resorption and delineates its underlying mechanisms. In vitro experiments reveal that KR notably inhibits osteoclastogenesis and reduces the expression of osteoclastic markers. Additionally, KR decreases the levels of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α, while downregulating NADPH oxidase 1 (NOX1) and Kelch-like ECH-associated protein 1 (Keap1) to diminish ROS production. Furthermore, KR activates the nuclear factor erythroid 2-related factor 2 (Nrf2), which enhances the activity of heme oxygenase-1 (HO-1) and catalase (CAT), facilitating the clearance of excess ROS. The compound also hinders osteoclast formation and functionality by inhibiting the PI3K/AKT/GSK-3β signaling pathway. Lentiviral knockdown of CAT can partially reverse these effects of KR. Meanwhile, in vivo experiments indicate that KR effectively mitigates bone loss in an LPS-induced inflammatory bone resorption model. In summary, KR is a promising new star in breaking through the limitations of previous drugs and treating inflammatory bone resorption.

Time-restricted feeding improves metabolic syndrome by activating thermogenesis in brown adipose tissue and reducing inflammatory markers

BackgroundObesity and metabolic syndrome (MetS) have become increasingly significant global health issues. Time-restricted feeding (TRF), as a novel dietary intervention, has garnered attention in recent years. However, there is limited research focusing on the effects of TRF on energy expenditure and systemic low-grade inflammation. This study aims to investigate the impact of TRF on weight management, glucose metabolism, insulin resistance, and lipid metabolism in male C57BL/6J mice, particularly in the context of metabolic disorders induced by a high-fat diet (HFD).MethodsC57BL/6J mice were divided into two groups: a normal diet (ND) group and a high-fat diet (HFD) group. The study duration was 12 weeks. Key parameters observed included body weight, glucose tolerance (via glucose tolerance tests), insulin resistance (HOMA-IR), and insulin secretion under glucose stimulation. Additionally, liver tissue was subjected to Oil Red O staining to assess lipid accumulation, and white and brown adipose tissues were stained with hematoxylin and eosin (HE) to evaluate adipocyte size. The expression of hepatic lipogenesis-related genes (Srebp-c, Chrebp, Fasn, and Acc1) and thermogenic genes in brown adipose tissue (UCP1 and PGC-1α) were also measured. Furthermore, temperature changes in the interscapular brown adipose tissue (BAT) were monitored.ResultsIn the ND group: TRF improved insulin resistance and reduced circulating levels of the pro-inflammatory cytokine IL-6, with a slight reduction in body weight.In the HFD group: TRF significantly mitigated weight gain, improved glucose tolerance and insulin resistance, and enhanced insulin secretion under glucose stimulation. Additionally, TRF reduced hepatic steatosis by downregulating the expression of lipogenesis-related genes in the liver. TRF also increased thermogenesis by upregulating the expression of thermogenic genes (UCP1 and PGC-1α) in BAT, while lowering serum levels of pro-inflammatory cytokines IL-6 and TNF-α, though IL-1β levels remained unchanged.ConclusionThis study demonstrates that TRF can activate thermogenesis in brown adipose tissue and reduce inflammation maker, leading to an improvement in hepatic steatosis and a reduction in white adipose tissue accumulation. These findings suggest that TRF may be a promising intervention for mitigating metabolic disturbances associated with obesity and metabolic syndrome. The study provides mechanistic insights into the beneficial effects of TRF, highlighting its potential in modulating lipid metabolism and exerting anti-inflammatory effects.

Assessment of Lactobacillus acidophilus (L. acidophilus) therapeutic and prophylactic role in rats experimentally infected with Blastocystis subtype 3 (ST3).

Blastocystis, an eukaryote, inhabits the intestinal tract of humans and animals worldwide. Lactobacillus acidophilus (L. acidophilus), a probiotic, has been reported to be effective against blastocystosis. The present study evaluated the therapeutic and prophylactic effectiveness of L. acidophilus compared to metronidazole (MTZ) in rats experimentally infected with Blastocystis subtype 3 (ST3). Four groups of Blastocystis ST3-infected rats received MTZ, L. acidophilus, both MTZ and L. acidophilus, or prophylactic L. acidophilus. Non-infected and infected control groups were included. The effectiveness of treatment and prevention was evaluated using parasitological monitoring, histopathological examination, immunohistochemical staining for TNF-α and IgA expression, and immunological testing for TNF-α and IL-10. In the L. acidophilus-treated group, a 76% reduction in the mean fecal parasitic count and a 67% clearance in the intestinal wash were achieved aligning closely with outcomes observed in the MTZ-treated group. An immunomodulatory impact via upregulation of the anti-inflammatory cytokine IL-10 and downregulation of the pro-inflammatory cytokine TNF-α was demonstrated as well. Combined administration of MTZ and L. acidophilus exhibited superior efficacy with a 99% decrease in the mean fecal parasitic count and a 98% decrease in the intestinal fluid parasitic count. Additionally, the lowest TNF-α and the highest IL-10 serum levels. Prophylactic use of L. acidophilus for 7 days neither prevented infection nor reduced its severity. Furthermore, no significant differences were detected in serum levels of TNF-α and IL-10 following post-prophylaxis andpost-treatmentwith L. acidophilus. Accordingly, L. acidophilus might be recommended as an adjuvant treatment alongside MTZ for improved efficacy against blastocystosis.

P0068 Distribution of α4β7 integrin and MAdCAM-1 expression according to the location of intestinal inflammation in a murine model of spontaneous chronic enterocolitis

Abstract Background The selective blockade of α4β7 integrin is a treatment for inflammatory bowel disease (IBD), with different efficacy according to the location of inflammation along the gastrointestinal tract. A more pronounced effect has been reported in patients with predominant colonic inflammation. It may be attributed to differences in the regional expression of adhesion molecules. This study aimed to investigate the expression of α4β7 integrin and mucosal addressin cell adhesion molecule-1 (MAdCAM-1) in colon, ileum, and mesentery, using IL-10 knockout (KO) mice as a model of spontaneous chronic enterocolitis. Methods At 20 weeks, the colon, ileum, and mesentery were extracted from IL-10KO and C57BL/6 wild-type (WT) mice. Protein levels of α4β7 integrin and MAdCAM-1 were determined by Western blot in the different tissues. Additionally, pro-inflammatory cytokines (TNF-α and IFN-γ) and epithelial adhesion molecules (Zonula Occludens-1, ZO-1; E-cadherin and Mucin 2, Muc2) expression in the ileum, as well as adiponectin expression in the mesentery, were analyzed using real-time PCR. Results Analysis demonstrated a significant increase in gene expression of the pro-inflammatory cytokines TNF-α and IFN-γ in the ileum of IL-10 KO mice compared with WT mice (p&amp;lt;0.01). In addition, IL-10KO mice exhibited reduced expression of ZO-1, E-cadherin, and Muc2 in the ileum relative to WT mice (p&amp;lt;0.001). IL-10 deficiency also led to a marked decrease in adiponectin expression in the mesentery (p&amp;lt;0.05). We then analyzed the protein level of MAdCAM-1 and α4β7 integrin according to anatomic region. Remarkably, the protein analysis revealed significantly higher levels of MAdCAM-1 (p&amp;lt;0.01) and α4β7 integrin (p&amp;lt;0.05) in the colon of IL-10 KO mice compared to WT. In the ileum, while MAdCAM-1 levels remained unchanged, α4β7 integrin expression was increased in IL-10KO mice (p&amp;lt;0.01). No significant differences in MAdCAM-1 or α4β7 integrin levels were observed in the mesentery between IL-10KO and WT mice Conclusion IL-10-deficient mice develop inflammation in the ileum, accompanied by a reduction in adiponectin production in the mesentery. In this setting, both α4β7 integrin and MAdCAM-1 showed increased expression in the colon, which was not observed in the ileum or mesentery, despite the inflammation. This differential regional expression pattern of adhesion molecules may explain the variation in the efficacy of anti-integrin therapies. Characterizing these patterns in patients with IBD could help identify those who would benefit most from these treatments.

DOP076 Translational and preclinical study validating anterior gradient 2 as a novel epithelial therapeutic target in Inflammatory Bowel Disease (IBD) involved in inflammation, fibrosis, and intestinal barrier integrity

Abstract Background Anterior Gradient 2 (AGR2) is an endoplasmic reticulum (ER) resident protein involved in protein folding, predominantly expressed in mucin-secreting epithelial cells. Under stress, AGR2 is secreted into the extracellular milieu (eAGR2), where it disrupts epithelial integrity, attracts monocytes, and activates fibroblasts. We explored the relationship between AGR2 tissue expression in IBD patients and the efficacy of a neutralizing anti-eAGR2 antibody (TH-009) in murine models of colonic inflammation and fibrosis Methods AGR2 expression was analyzed by immunohistochemistry on ileal and colonic biopsies from IBD patients and controls. In mice, the efficacy of TH-009 was evaluated in models of colitis (3% DSS) and digestive fibrosis (3 cycles of 1.5% DSS) Results 132 ileal and colonic endoscopic biopsies from 66 patients with Crohn’s disease (CD), 50 patients with ulcerative colitis (UC), and 16 controls, and 96 surgical samples from 11 CD patients, 24 UC patients, and 11 controls were included. AGR2 expression correlated with mucosal inflammation (R² = 0.416, p&amp;lt;0.0001) and tissue damage (R² = 0.329, p&amp;lt;0.0001). All patients with mucosal inflammation showed AGR2 overexpression, while 38% of histologically healed patients had high AGR2, with a relapse rate at 18 months three times higher than those whose AGR2 levels were comparable to controls.In a murine acute colitis model (3% DSS), intravenous (IV) treatment with TH-009 at doses of 5 and 20 µg showed an improvement in weight (p&amp;lt;0.005 and p=0.0001, respectively), in the global colitis DAI score (p=0.002 and p=0.001), and in the histological colitis score (p=0.04 for 5 µg) compared to controls. Intraperitoneal and subcutaneous administration were also effective. There was a significant reduction in tissue infiltration by macrophages (p&amp;lt;0.01) and T lymphocytes (p&amp;lt;0.001), in myeloperoxidase activity, and in the tissue and serum expression of pro-inflammatory cytokines TNFα and IL-6. TH-009-treated mice restored intestinal permeability, measured by FITC-Dextran blood passage, to levels comparable to unexposed DSS mice, unlike DSS-only exposed mice (p&amp;lt;0.01). In a fibrosis model (3 cycles of 1.5% DSS), preventive and curative treatment with TH-009 reduced digestive fibrosis by 49% (p&amp;lt;0.0001), and reduced submucosal fibrosis thickness by 59% (p&amp;lt;0.0001). There was a significant decrease in the colonic expression of fibrosis-related genes (Collagen A1, αSMA, fibrinogen, and TGFβ). Conclusion AGR2 is a novel therapeutic target in IBD, correlating with inflammation and complications. Its expression is not affected by major therapies and is overexpressed in 40% of patients with deep histological healing. TH-009 improves inflammation, reverses fibrosis, and restores epithelial barrier functions.

P0131 Characterisation of a human multi-organ-on-chip model to unravel gut-blood-brain communication

Abstract Background Persistent fatigue is a frequently reported symptom among patients with Inflammatory Bowel Disease (IBD), significantly diminishing quality of life. Due to the lack of understanding how IBD-associated fatigue originates, currently available treatments do not efficiently tackle this symptom. Developing targeted therapies requires tools to unravel the mechanisms behind central nervous system dysfunction in response to chronic intestinal inflammation. We are therefore establishing and characterising a human multi-organ-on-chip model to recapitulate key physiological elements of gut-brain communication via the systemic circulation. Methods In our model, a gut-blood barrier, comprised of Caco-2 (enterocytes) and HT29-MTX cells (goblet cells), is connected to a blood-cerebrospinal fluid (CSF) barrier, composed of HIBCPP and iHCPEnC cells (choroid plexus epithelial and endothelial cells respectively), and to a brain compartment (SH-SY5Y, neurons) via recirculation of cell culture medium. Gut inflammation was mimicked by the addition of the pro-inflammatory cytokines TNFα and IFNγ to the intestinal epithelial cells. Barrier integrity was measured by transepithelial/endothelial resistance (TEER) and by paracellular permeability assays with 4 kDa FITC-dextran. In addition, we performed a bead-based immunoassay to measure secreted IL-8 levels and RT-qPCR to analyse differential gene expression. Results The gut-blood barrier under low perfusion (3,6*10-4 mPa) exhibited increased expression of differentiation marker genes compared to static culture, such as LYZ, SOX9 and CYP3A4, indicating an improvement of cell differentiation. Moreover, low perfusion induced the formation of 3D villi-like structures, mucus production and the reduction of barrier strength towards more physiological relevant levels (TEER: 81,6±44,8 Ω.cm2; Papp: 1.10-5±4.10-6 cm/s). The gut-blood barrier was responsive to the addition of TNFα and IFNγ and high IL-8 levels could be measured in the circulatory blood flow. The barrier properties of the blood-CSF barrier under low perfusion (endothelial: 2,52 mPa, epithelial: 4,6*10-4 mPa) were not significantly different from a static setup, but less variance was observed in TEER. Neurons differentiated with BDNF and retinoic acid in our model displayed long projections and expressed mature neuronal marker genes (SYP, SNAP25, TUBB3). Conclusion We have successfully established and characterised each organ compartment of the gut-blood-brain model. Moreover, we have proven that low perfusion improves the physiological relevance of our model, especially of the gut-blood barrier. We are now exploring phenotypical and transcriptional alterations across the blood-CSF barrier and the brain compartment when gut inflammation is triggered.

Maternal exposure to chronic, low-dose nonylphenol in zebrafish: Disruption of ovarian health, reproductive function, and embryogenesis.

Nonylphenol (NP), a non-ionic surfactant and potent endocrine disruptor, is known for its environmental persistence, biotic accumulation potential and toxicity. Nonetheless, mechanisms underlying NP modulation of female fertility with potential impact on embryogenesis in the unexposed offspring remain elusive. This study investigates the effects and toxic mechanisms of maternal exposure to NP at varying concentrations (50 and 100μg/L) on zebrafish (Danio rerio), specifically focusing on ovarian health, reproductive parameters, and early developmental potential in the F1 generation. Our findings indicate a higher accumulation of NP in the ovaries compared to muscle tissue. Further, chronic (28 days) NP exposure promotes ovarian reactive oxygen species (ROS) accumulation, activates the MAPK (JNK, p38 MAPK, ERK1/2) pathways, AP-1 induction, and elevated expression of pro-inflammatory cytokines (Tnf-α, Il-1β, Il-6) triggering inflammation. Besides, heightened follicular atresia in NP-treated ovaries relates to increased Bax/Bcl2 ratio, cleaved caspase 3 and Parp1 activation prompting apoptosis. While it showed higher affinity to zebrafish ERα (in silico analysis), NP exposure in vivo promotes a robust increase in ovarian ERα but abrogated ERβ expression and a significant alteration in fshr and lhcgr transcripts. While attenuated StAR and P450 aromatase expression at both mRNA and protein levels and reduced igf3 expression reveal impaired ovarian microenvironment, NP-induced dysregulated NO/NOS/cyclooxygenase signaling and attenuation of hCG-induced p34cdc2 activation and oocyte maturation correspond well with decreased fecundity and fertilization efficiency. Intriguingly, maternal exposure to NP resulted in delayed embryogenesis, developmental aberrations, and reduced hatching rates in the unexposed offspring, risking F1 generation. Collectively, this study provides mechanistic insights into the detrimental influence of maternal exposure to NP on ovarian dysfunction, reproductive insufficiency and embryotoxicity.

Protective effects of hydatid cyst fluid on inflammation and tissue damage in rat model of type 1 diabetes.

Cystic echinococcosis, caused by Echinococcus granulosus, is a zoonotic disease with immunomodulatory properties attributed to hydatid cyst fluid (HCF). Given the immune-modulating and anti-inflammatory properties of HCF observed in other contexts, its potential therapeutic effects in diabetes remain unexplored. This study aimed to investigate the potential therapeutic effects of HCF on glycemic control, inflammatory cytokines, and tissue histopathology in a streptozotocin (STZ)-induced model of type 1 diabetes. Twenty male rats were randomly divided into four groups (n = 5): a healthy control group, a hydatid cyst group that received three intraperitoneal injections of HCF at two-week intervals, a diabetic group that received a single intraperitoneal dose of STZ to induce diabetes, and a hydatid cyst + diabetic group (HCF + STZ) that received both HCF treatment and STZ administration. Serum glucose levels, inflammatory cytokines (TNF-α, IL-1β, and IL-10), and histopathological changes in pancreatic and renal tissues were analyzed. The HCF + STZ group demonstrated a significant reduction in serum glucose levels compared to the STZ-only group. Pro-inflammatory cytokines TNF-α and IL-1β were significantly decreased in HCF-treated diabetic rats, while the anti-inflammatory cytokine IL-10 was partially restored. Histopathological examination revealed severe pancreatic islet atrophy and renal degeneration in the diabetic group, which were markedly alleviated in the HCF + STZ group. These findings suggest that HCF's immunomodulatory and anti-inflammatory properties may mitigate hyperglycemia and inflammatory responses in type 1 diabetes, warranting further investigation into its mechanisms and clinical applications.

Design, synthesis, and anti-inflammatory activity of 2H-1,4-benzoxazin-3(4H)-one derivatives modified with 1,2,3-triazole in LPS-induced BV-2 cells.

Given the potent anti-inflammatory properties of the 1,2,3-triazole structure and the wide use of 2H-1,4-benzoxazin-3(4H)-one in developing treatments for neurodegenerative diseases, a series of 2H-1,4-benzoxazin-3(4H)-one derivatives were synthesized by introducing a 1,2,3-triazole moiety. Screening for anti-inflammatory activity in microglial cells revealed that compounds e2, e16, and e20 exhibited the most promising effects without significant cytotoxicity. These compounds effectively reduced LPS-induced NO production and significantly decreased the transcription levels of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α. Furthermore, they downregulated the transcription and protein levels of the inflammation-related enzymes iNOS and COX-2 in response to LPS stimulation. To further investigate the anti-inflammatory mechanisms of these derivatives in microglia, the intracellular ROS levels and the activation of the Nrf2-HO-1 signaling pathway were analyzed. The results indicated that the 2H-1,4-benzoxazin-3(4H)-one derivatives significantly activated the Nrf2-HO-1 pathway, reduced LPS-induced ROS production, and alleviated microglial inflammation. Molecular docking studies suggested that compounds e2, e16, and e20 could interact with Nrf2-related binding sites, preventing its degradation by Keap1. Additionally, acute toxicity tests in mice demonstrated that compound e16 exhibited favorable safety.

Modulation of Intestinal Inflammation and Protection of Dopaminergic Neurons in Parkinson’s Disease Mice through a Probiotic Formulation Targeting NLRP3 Inflammasome

Emerging evidence highlights the significance of peripheral inflammation in the pathogenesis of Parkinson’s disease (PD) and suggests the gut as a viable therapeutic target. This study aimed to explore the neuroprotective effects of the probiotic formulation VSL#3® and its underlying mechanism in a PD mouse model induced by MPTP. Following MPTP administration, the striatal levels of dopamine and its metabolites, as along with the survival rate of dopaminergic neurons in the substantia nigra, were significantly reduced in PD mice. MPTP also significantly increased the mRNA expression of pro-inflammatory cytokines TNF-α and IL-1β, while reducing anti-inflammation mediators, like glia cell line-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF) in the striatum. These pathological changes were notably mitigated by VSL#3® treatment, suggesting its neuroprotective and anti-inflammatory effects in the brain. Additionally, VSL#3® significantly lowered the circulating levels of pro-inflammatory cytokines, and reduced TNF-α and IL-1β mRNA expression in the liver, indicating an inhibition of cytokine transfer. In the intestine, the probiotic treatment markedly decreased the mRNA expression of pro-inflammatory cytokines, (TNF-α, IL-1β, IL-6 and IL-17), and the other two key components of the NLRP3 inflammasome, caspase-1 and NLRP3, demonstrating an inhibition of VSL#3® on gut NLRP3 inflammasome. VSL#3® exerts neuroprotective effects in PD mice through the suppression of intestinal inflammation, particularly inhibiting the intestinal NLRP3 inflammasome. This study supports the therapeutic potential of targeting intestinal inflammation and utilizing probiotics in PD treatment.

Resveratrol Anti-inflammatory Effect against Palmitate-induced Cytotoxicity in Raw 264.7 Macrophages.

Resveratrol (RES) is a phytochemical bioactive compound with suggested therapeutic benefits. The current work aimed to evaluate the anti-inflammatory effect of RES against palmitate (PA) induced lipotoxicity in raw 264.7 macrophages cell line. The cells viability was assessed by lactate dehydrogenase assay. Then the effects of RES and PA on nitric oxide (NO), triglyceride (TG) content, and cytokines release were studied. The effect of RES and PA on the treated cells bioenergetics and redox status was evaluated via different assays Results: The results showed that at doses of 10 and 20μM, RES dramatically increased the vitality of PA-exposed macrophages with dramatic significant decrease in the release the proinflammatory cytokines TNF-α, MHGB-1, IL-1β, and IL-6 and their coding genes expression with marked improvement in the cells phagocytic capacity. In addition, RES dramatically lowered the levels of NO and TG in PA-stimulated macrophages. In addition, PA markedly decreased mitochondrial complexes I and III activities with decreased mitochondrial membrane potential and lowered ATP production with induction of oxidative stress. RES was shown to mitigate the effect of PA on macrophages bioenergetics and the oxidative damage and counteracted PA effect on genes linked to oxidative damage, such as Nrf2, Ho-1, NF-κB p65, SOD1, and SOD2. RES could reduce PA-induced lipotoxicity in macrophages via enhancing their viability and counteracting the excess release of cytokines through alleviating PAinduced bioenergetic disruption and oxidative damage with a suggested positive impact of RES on obesity related illnesses caused by triggered cellular inflammation.

L-selectin Promotes Migration, Invasion and Inflammatory Response of Fibroblast-Like Synoviocytes in Rheumatoid Arthritis via NF-kB Signaling Pathway.

Rheumatoid arthritis (RA) is a chronic, systemic autoimmune disease characterized by chronic inflammation of the synovium and progressive joint damage. Fibroblast-like synoviocytes (FLSs) exhibit excessive proliferative and aggressive phenotypes and play a major role in the pathophysiology of RA. Previous studies have confirmed the pathologic role of L-selectin in cell adhesion and migration. In rheumatoid arthritis models, L-selectin regulates leukocyte homing, which leads to joint inflammation. Moreover, in L-selectin knockout mice, there is a reduction in joint inflammation. However, the associations of L-selectin with FLSs in RA remain unclear. This study aims to reveal the effect of L-selectin on RA-FLSs and to investigate the molecular mechanism of L-selectin in RA. Our findings indicated that L-selectin was significantly expressed in RA synovial tissues and RA-FLSs. L-selectin silencing reduced RA-FLSs migration and invasion and attenuated the secretion of pro-inflammatory cytokines TNF-α, IL-1β and IL-6 in vitro. Moreover, investigations into mechanisms revealed that L-selectin activated the nuclear factor kappa-B (NF-κB) signaling pathway while blocking this signaling pathway could compromise the effects of L-selectin. Finally, in vivo experiments with a collagen-induced arthritis rat model revealed that silencing L-selectin alleviated inflammatory infiltration of the synovium and cartilage destruction, and validated the NF-κB signaling pathways findings observed in vitro. In summary, we show that L-selectin enhances the migration and invasion of RA-FLSs through the activation of NF-κB signaling pathways, ultimately worsening the progression of RA.

Detrimental effects of chronic sugar-sweetened carbonated soft drink consumption on inflammatory response and the size of apical periodontitis: An animal-based study.

This study aimed to evaluate the effects of chronic consumption of two sugar-sweetened carbonated soft drinks - one containing caffeine (Coca-Cola®) and one without (Sprite®) - on the progression of periapical lesions and the levels of pro-inflammatory cytokines in rats. Twelve Wistar rats were divided into three groups (n = 4): Control group, Coca-Cola group and Sprite group. The rats in Coca-Cola and Sprite groups were given ad libitum access to their respective soft drinks for 3 months, while the Control group received filtered water. After 2 months of consumption, the pulps of the lower left first molars were exposed for 28 days to induce periapical lesions. Following euthanasia, the jaws were removed, and the periapical lesions were assessed using micro-computed tomography imaging. Blood samples were collected to analyse the pro-inflammatory cytokines IL-1β, IL-6, IL-2, IL-17 and TNF-α via Luminex assay. Non-parametric data were analysed using the Kruskal-Wallis test followed by Dunn's test, while parametric data were analysed using one-way ANOVA followed by Tukey's test (p < 0.05). Both the Coca-Cola and Sprite groups exhibited periapical lesions with significantly greater volume and diameter compared to the Control group (p < 0.05). Additionally, both soft drink groups had significantly higher levels of IL-1β, IL-6 and IL-2 compared to the Control group (p < 0.05). The Sprite group displayed significantly higher levels of IL-1β than the Coca-Cola group (p < 0.05), while the Coca-Cola group showed significantly elevated TNF-α levels compared to both the Control and Sprite groups (p < 0.05). No significant differences in IL-17 levels were observed among the groups (p > 0.05). The chronic consumption of sugar-sweetened carbonated soft drinks, regardless of caffeine content, has detrimental effects on the inflammatory response and progression of apical periodontitis in rats.

Uterine Myometrial Distension Augments the Production of Angiogenic and Proinflammatory Factors

We recently found that myometrial distension stimulates maternal uterine vascular remodeling, and hypothesized that this may be a previously unrecognized mechanism for inducing arterial growth during pregnancy. The aim of this study was to further characterize a recently developed surgical model in which medical-grade silicone was infused into one uterine horn of a non-gravid rat to induce acute myometrial stretch, followed by an additional, gradual distension due to the secretion of an exudate into the uterine lumen. Our objectives were to better understand the effects of stretch on the myometrium and to look for the expression of proangiogenic and proinflammatory factors that may stimulate uterine vascular remodeling. Morphometric analysis showed hypertrophy of the uterine corpus that was primarily due to axial growth since the myometrial cross-sectional area was unchanged due to a thinning of the uterine wall secondary to stretch. This finding was supported by significantly increased myometrial smooth muscle cell mitosis. There was also an increase in the concentration of myometrial elastin but not collagen. The analysis showed modest increases in neutrophils, activated and unactivated macrophages, and the proinflammatory cytokines RANTES, MIP-3α, GRO-KC, and TNFα. The most dramatic change was the extremely high level of VEGF in the exudate, which was increased &gt;900× above circulating levels.