Expression Levels Of IL-1β Research Articles

Evaluation of the immunomodulatory effect of siRNA Nup98.1 и Nup205.1 during influenza infection using in vivo models

BACKGROUND: Influenza infection is one of the most pressing global health problems. The problems of the formation of viral resistance to existing drugs and the immunosuppressive effect induced by the influenza virus are currently acute. A promising solution seems to be the creation of drugs whose action is based on inhibition of the activity of cellular genes through RNA interference. AIM: Evaluate the anti-influenza and immunomodulatory effect of small interfering RNAs directed to cellular genes in an in vivo model. METHODS: The study used the influenza virus strain A/California/7/09 (H1N1), BALB/c mouse. Small interfering RNA was administered and animals were infected intranasally. Changes in the cytokine profile and viral activity were assessed using molecular genetic and virological methods. RESULTS: In the lungs of mice with intranasal administration of all siRNA complexes, a decrease in the expression level of IL-1β and TNF-α was noted, but an increase in the expression of IL-1β and IL-6 in washings from the upper respiratory tract (URT) was noted. When using siRNA Nup98.1 and Nup98.1 / Nup205.1, a reliable increase in the expression of IL-10 in the lungs and URT by the third day was observed. Changes in expression were also noted in relation to TGF-β1. Reliable differences in the expression of pro- and anti-inflammatory cytokines were obtained. Based on this, the most active immunomodulatory effect of the used small interfering RNA complexes is shown in the upper respiratory tract and lungs, at the sites of their direct administration. CONCLUSION: The study presented data that small interfering RNAs directed to one or more cellular genes in an in vivo model can reliably lead to a decrease in viral reproduction, a change in the cytokine profile, as well as an increase in the survival rate of laboratory animals.

Network Pharmacology and Experimental Validation to Reveal the Pharmacological Mechanisms of Gynostemma pentaphylla against Acute Pharyngitis.

Acute pharyngitis (AP) is a prevalent ailment. Gynostemma pentaphylla (GP), a traditional Chinese medicine (TCM), may treat AP due to its anti-tumor and anti-inflammatory properties, but this remains unexplored. This study utilized the TCMSP and Swiss Target Prediction databases to analyze GP's chemical composition and target proteins. The Genecards database was used to identify targets relevant to AP. A PPI network diagram of drug-disease intersection targets was created using the STRING database, and Cytoscape was utilized to create a network visualization diagram of "GP active components-targets-AP" in order to determine key active components of GP in treating AP. Gene ontology (GO) and biological pathway (KEGG) enrichment analyses were conducted on targets in the David database. Molecular docking verification of key targets and components was performed using AutoDock Vina software. In animal experiments, a rat model of AP was induced by a 15% concentrated ammonia solution, and HE staining was conducted to observe histopathological changes in the rat pharynx after intragastric administration of Houyanqing. ELISA was used to detect expression levels of serum interleukin-1-beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor (TNF-α). A total of 18 active ingredients were screened from GP, among which Ruvoside _ qt, Rhamnazin, 3 ' -methyleriodictyol, and sitosterol were five key active ingredients. The key targets involved EGFR, STAT3, MAPK3, SRC, AKT1, etc. KEGG enrichment analysis showed that GP mainly acted on Pathways in cancer, P13K-AKT signaling Pathways, JAK-STAT signaling pathways, and other signaling pathways. Molecular docking results showed that four core compounds and five key targets met the energy matching. Animal experiments showed that compared with the normal group, the expression levels of IL-1β, IL-6, and TNF-α in the AP model group were significantly up-regulated (P < 0.05). In addition, compared with the model group, intragastric administration of the dexamethasone group and gypenosides group could alleviate the up-regulation of inflammatory factors in model rats, and the levels of IL-1β, IL-6, and TNF-α were decreased (P < 0.05). This study predicted the possible targets of GP in the treatment of AP through network pharmacology. The results suggest that gypenosides may inhibit the expression of inflammatory factors by regulating Pathways in cancer, P13K-AKT, and JAK-STAT signaling pathways to treat AP.

Protective Role of Nano-encapsulated Bifidobacterium breve, Bacilllus coagulans, and Lactobacillus plantarum in Colitis Model: Insights Toward Propagation of Short-Chain Fatty Acids and Reduction of Exaggerated Inflammatory and Oxidative Response.

Irritable bowel disease (IBD), also known as ulcerative colitis and Crohn's disease, is a chronic inflammatory disorder affecting millions of people worldwide. Herein, nano-encapsulated multi-strain probiotics formulation, comprising Bifidobacterium breve DSM24732 and B. coagulans SANK 70258 and L. plantarum DSM24730 (BBLNPs) is used as an effective intervention technique for attenuating IBD through gut microenvironment regulation. The efficacy of the prophylactic role of BBLNPs in alleviating injury induced by dextran sulfate sodium (DSS) was evaluated by assessing oxidative and inflammatory responses, levels of short-chain fatty acids (SCFAs) and their regulation on GPR41/43 pathway, expression of genes related to tight-junctions and autophagy, immunohistochemistry of IL1β and GPR43, and histological examination of inflamed colonic tissue. The severity of clinical signs and paracellular permeability to FITC (fluorescein isothiocyanate)-labeled dextran was significantly decreased after BBLNP treatment. Reduction of oxidative stress-associated biomarkers (MDA, ROS, and H2O2) and acceleration of antioxidant enzyme activities (SOD, CAT, and GSH-Px) were noted in the BBLNP-treated group. Subsiding of inflammatory markers (TNF-α, IL-18, IL-6, TRL-4, CD-8, NLRP3, and caspase 1) and upregulation of tight-junction-related genes (occludin and JAM) was detected in BBLNPs. Administration of BBLNPs remarkably resulted in a higher level of SCFAs which parrel with colonic upregulation of GPR41 and GPR43 expression compared to DSS-treated rats. Notable modulation of autophagy-related genes (p62, mTOR, LC3, and Beclin-1) was identified post BBLNP treatment. The mRNA expressions of p62 and mTOR were significantly downregulated, while LC3 and Beclin-1 were upregulated after prophylactic treatment with BBLNPs. Immune-stained labeled cells showed lower expression of IL-1β and higher expression levels of GPR43 in BBLNPs compared to the DSS-induced group. The intestinal damage caused by DSSwas effectively mitigated by oral BBLNP treatment, as supported by the restoration of healthy colonic tissue architecture. The findings suggest that BBLNPs have a promising avenue in the remission of IBD by modulating inflammation, oxidative stress, microbial metabolites such as SCFAs, and autophagy.

Coenzyme Q10 and its liposomal form prevent copper cardiotoxicity by attenuating oxidative stress, TLR-4/NF-κB signaling and necroptosis in rats.

Copper (Cu) is an essential element involved in numerous biochemical, metabolic and cellular processes. Excessive exposure to the pesticide copper sulfate (CuSO4) was associated with toxic effects. This study aims to evaluate the efficacy of Coenzyme Q10 (CoQ10) and its liposomal form (L-CoQ10) against myocardial injury induced by CuSO4, pinpointing the involvement of redox imbalance, TLR-4/NF-κB signaling and apoptosis. Cardiac injury in rats was induced by daily oral doses of CuSO4 for 7 days, the rats were treated orally with either CoQ10 or L-CoQ10 concurrently with CuSO4 for 7 days. Elevated serum cTnI, CK-MB and LDH were observed in CuSO4-intoxicated animals. Additionally, cellular antioxidant biomarkers were decreased and the expression levels of cardiac MDA, TLR-4, NF-κB, IL-6, IL-1β, and TNF-α were upregulated. CoQ10 and L-CoQ10 prevented myocardial injury and decreased the levels of both MDA and pro-inflammatory cytokines. CoQ10 and L-CoQ10 enhanced antioxidant capacity and Bcl-2, and downregulated caspase-3, Bax, p53, RIP3, MLKL, caspase-8 and TLR-4/NF-κB signaling. In conclusion, CoQ10 and L-CoQ10 effectively prevent CuSO4 cardiotoxicity in rats. Attenuation of redox imbalance, TLR-4/NF-κB signaling, pro-inflammatory response, and necroptosis along with enhancement of antioxidant response mediated their cardioprotective efficacy. CoQ10 could be valuable in protecting people vulnerable to Cu toxicity.

Role of TNFRSF12A in cell proliferation, apoptosis, and proinflammatory cytokine expression by regulating the MAPK and NF-κB pathways in thyroid cancer cells.

Tumor necrosis factor receptor superfamily member 12A (TNFRSF12A) has been reported to be upregulated in thyroid cancer (THCA). However, the role and mechanism of TNFRSF12A in THCA remain largely unknown. TNFRSF12A expression in THCA samples was analyzed using bioinformatics analysis. CCK-8, EdU incorporation assay, TUNEL, and caspase-3 activity assay was used to detect cell proliferation and apoptosis in THCA cells. Correlated genes of TNFRSF12A were identified using LinkedOmics database and subjected to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Western blot analysis was performed to determine proliferating cell nuclear antigen (PCNA), cyclin D1 (CCND1), Bax, and Bcl-2 expression and to analyze the effect of TNFRSF12A on mitogen-activated protein kinase (MAPK) and nuclear factor-kappaB (NF-κB) pathways. Results showed that TNFRSF12A was increased in THCA tissue samples and cells. KEGG analysis showed that correlated genes of TNFRSF12A were significantly enriched in MAPK and NF-κB signaling pathways. Moreover, TNFRSF12A knockdown inactivated the MAPK and NF-κB signaling pathways in THCA cells. TNFRSF12A silencing alone or combined with inhibitor of ERK (PD98059), JNK (SP600125), p38 (SB203580), or NF-κB (Bay 11-7082) impeded cell proliferation and reduced PCNA and CCND1 expression in THCA cells. Meanwhile, TNFRSF12A knockdown alone or combined with PD98059, SP600125, SB203580, or Bay 11-7082 facilitated cell apoptosis, increased caspase-3 activity, downregulated Bcl-2 expression, and upregulated Bax expression in THCA cells. TNFRSF12A knockdown alone or combined with PD98059, SP600125, SB203580, or Bay 11-7082 also decreased the expression levels of proinflammatory cytokines IL-1β, IL-6, and IL-8 in THCA cells. On the contrary, TNFRSF12A overexpression showed an opposite effect. Treatment with PD98059, SP600125, SB203580, or Bay 11-7082 reversed the effects of TNFRSF12A overexpression on cell proliferation, apoptosis, and proinflammatory cytokine expression. In conclusion, the effects of TNFRSF12A on proliferation, apoptosis, and proinflammatory cytokine expression in THCA cells were regulated by the MAPK and NF-κB pathways.

Tryptophan ameliorates soybean meal-induced enteritis via remission of oxidative stress, mitophagy hyperactivation, and apoptosis inhibition in hybrid yellow catfish gut (Pelteobagrus fulvidraco ♀ × Pelteobagrus vachelli ♂)

In this study, the effects of Trp on the growth performance and intestinal health of hybrid yellow catfish suffering from soybean meal-induced enteritis (SBMIE) were investigated. A total of 600 individuals (initial body weight: 6.47 ± 0.01 g) were randomly divided into 12 cages, each with a water volume of 0.9 m3 and containing 50 individuals. Four different diets were used to feed hybrid yellow catfish: a fish meal-based diet (containing 40 % fishmeal, FM diet), a soybean meal-based diet (dietary replacement of 75 % FM with soybean meal, SBM diet), and two tryptophan-supplemented diets (soybean meal-based diet containing 0.5 % or 1.0 % of tryptophan, respectively, Trp1 diet and Trp2 diet), were used to feed hybrid yellow catfish. After 42 days of feeding, growth performance, oxidation and antioxidant indices, intestinal histomorphology, mitophagy and apoptosis were analyzed. The results revealed that the final body weight (FBW), weight gain rate (WGR), specific growth rate (SGR), feed conversion rate (FCR), viscerosomatic index (VSI), hepatosomatic index (HSI), and condition factor (CF) were unaffected by Trp (P > 0.05) compared with those of fish fed the SBM diet. The fish fed the SBM diet exhibited typical SBMIE, with significantly decreased T-AOC, T-SOD, CAT, and GSH-PX, and increased MDA, ROS, DAO and D-lactate (P < 0.05). The intestinal and mitochondrial morphologies were disrupted. Conversely, the decreased antioxidant capacity, increased oxidant stress and intestinal permeability were significantly reversed by dietary Trp, and the intestinal and mitochondrial structures were repaired (P < 0.05). Moreover, the protein and mRNA levels of ZO-1, Occludin, and Claudin-1, as well as the mRNA expression levels of tgf-β, il-10, caspase 3, caspase 9, fis1, mff, drp1, mfn1, opa1, and nrf1, were significantly higher in the dietary Trp groups compared to the SBM group (P < 0.05). Additionally, the apoptotic signals in the hindgut were significantly greater in the dietary Trp groups than in the SBM group. (P < 0.05). The mRNA expression levels of tnf-α, il-1β, beclin1, lc3a, lc3b, parkin, pink1, bnip3, and fundc1, as well as the protein level of LC3B, were significantly decreased by dietary Trp (P < 0.05). In conclusion, these results suggest that Trp can be used as a feed additive to alleviate oxidative stress, ameliorate intestinal epithelial injury, and reduce hyperactivated mitophagy and apoptosis resistance in the intestinal epithelium of hybrid yellow catfish subjected to SBMIE.

P2RX1-blocked neutrophils induce CD8+ T cell dysfunction and affect the immune escape of gastric cancer cells

BackgroundGastric cancer (GC) is one of the deadly malignancies of the gastrointestinal tract. Research has confirmed the linkage of P2RX1 with immune cell activation and tumor progression. This project focused on the impact of P2RX1 level in neutrophils on the efficacy of immune checkpoint inhibitor (ICI) treatment in GC. MethodsBlood samples from 23 GC patients eligible for camrelizumab treatment were collected. Flow cytometry was carried out to analyze the proportion of P2RX1 in neutrophils. IHC was utilized to detect the expression level of PD-L1. We also evaluated the chemotaxis ability of neutrophils using a Transwell system, assessed the viability and apoptosis rate of GC cells using CCK-8 and flow cytometry, measured the proportions of CD8+PD-1+ and CD8+GZMB+ cells, determined the expression levels of IL-6, TNFα, IFN-γ, IL-8, IL-12, IL-1β, and GZMB by utilizing enzyme-linked immunosorbent assay (ELISA), and examined the expression levels of P2RX1 and PD-L1 using western blot (WB). By establishing a xenograft mouse model, we studied the impact of P2RX1-blocked neutrophils on the efficacy of ICI treatment in the GC microenvironment. ResultsIn GC, clinical analysis revealed increased infiltration of P2RX1-lowly expressed neutrophil subsets and increased expression of PD-L1. In vitro experiments demonstrated that abnormal expression of P2RX1 affected neutrophil function. Furthermore, the blockage or knockdown of P2RX1 in neutrophils modulated CD8+ T cell function, promoting GC progression. In in vivo experiments, the blockage of P2RX1 in neutrophils inhibited the effectiveness of ICI treatment in the GC microenvironment. ConclusionThis project validated that the loss of P2RX1 in neutrophils induces CD8+ T cell dysfunction and affects the GC development, indicating that P2RX1 may be an accurate biomarker for predicting ICI response, thus providing a theoretical basis for the clinical application of ICI.

Bacillus coagulans alleviates intestinal barrier injury induced by Klebsiella pneumoniae in rabbits by regulating the TLR4/MyD88/NF-κB signalling pathway.

Probiotics effectively alleviate host diarrhoea, but the specific mechanism is not clear. Therefore, we explored the protective mechanism of Bacillus coagulans (BC) on intestinal barrier injury induced by Klebsiella pneumoniae (K. pneumoniae) in rabbits by HE, immunofluorescence and 16S rRNA. The results showed that BC pretreatment alleviated the changes in average daily gain, average daily feed intake and FCR caused by K. pneumoniae in rabbits. Moreover, BC alleviated the inflammatory cell infiltration, intestinal villus reduction, crypt deepening and goblet cell reduction caused by K. pneumoniae in rabbits. Further research revealed that BC improved the intestinal barrier by improving the mechanical barrier, chemical barrier, immune barrier and microbial barrier. Specifically, BC improved the intestinal mechanical barrier by improving the intestinal structure, increasing the protein expression of PCNA, increasing the number of goblet cells, and altering the expression of occludin, claudin-1 and ZO-1. BC improved the intestinal chemical barrier by regulating the expression of MUC1 and MUC2 and inhibited the TLR4/MyD88/NF-κB signalling pathway by altering the expression levels of the inflammatory factors IL-1β, IL-6 and TNF-α, thus optimizing the intestinal immune barrier. In addition, adding BC to the diet improved the intestinal microbial barrier of rabbits by reducing the abundance of harmful bacteria and increasing the abundance of beneficial bacteria. In summary, BC protects against K. pneumoniae-induced intestinal barrier damage by improving intestinal morphology, mitigating the inflammatory response and regulating the microbial composition. Among the pretreatments, the pretreatment effect of 1 × 106 CFU/g was the best. This study provides a theoretical basis for the use of BC to prevent and treat diarrhoea caused by K. pneumoniae in rabbits.

Low-intensity Pulsed Ultrasound Stimulation of the Intestine Improves Insulin Resistance in Type 2 Diabetes.

Ultrasound stimulation of internal organs and peripheral nerves has demonstrated promising potential in regulating blood glucose metabolism. This study aims to assess the effectiveness of low-intensity pulsed ultrasound stimulation (LIPUS) on intestine in improving insulin resistance with type 2 diabetes mellitus (T2DM). C57BL/6J mice, both normal and T2DM, were randomly divided into three groups: Control, T2D-sham, and T2D-LIPUS. The T2D-LIPUS group received LIPUS stimulation in the intestine. The parameters were as follows: 1 MHz frequency, 1.0 kHz pulse repetition frequency (PRF), 20% duty cycle, 100 mW/cm² intensity spatial average temporal average (ISATA), for 20 minutes per session, five days per week, over four weeks. Blood glucose analysis indicated that mice in the T2D-LIPUS group displayed significantly lower area under the curve (AUC) of glucose tolerance tests (GTT) and insulin tolerance tests (ITT) (p < 0.001), HOMA-IR (p < 0.001), and fasting serum insulin levels (p < 0.01) compared to the T2D-sham group. LIPUS treatment effectively lowered serum levels of IL-1β (p < 0.001) and TNF-α (p < 0.01) along with mRNA expression levels of IL-1β (p < 0.01) and IL-18 (p < 0.001) in the intestines of T2DM mice. Additionally, Western blot analysis revealed a reduction in the protein levels of NLRP3, caspase-1, and GSDMD-N in the intestinal tissues of mice treated with LIPUS. These findings suggest that LIPUS can reduce inflammation and cellular apoptosis, while improving insulin resistance by inhibiting the NLRP3/Caspase-1/GSDMD signaling pathway. This research introduces a novel, non-pharmacological approach for managing T2DM.

Molecular responses to clove and oregano essential oils are associated with reduced inflammation and improved gut barrier function in broiler chickens.

In vitro tests were conducted to characterize the host-mediated responses of chickens to Clove Essential Oil (CEO) and Oregano Essential Oil (OEO). Chicken macrophage cells (CMCs), chicken intestinal epithelial cells (IECs), quail muscle cells (QMCs), and chicken embryonic muscle cells (EMCs) were utilized in these assays. EMCs were collected from the 13-day-old embryo during egg incubation and all cell lines were seeded at 2 × 105/mL in a 24-well plate. In CMCs, an inflammatory response was induced by stimulating with 1.0 µg/mL of Lipopolysaccharide (LPS). To induce muscle cell differentiation, 0.5 % FBS was used in QMCs and 2.0 % FBS in EMCs. Three different concentrations (1.0, 10.0, and 100 µg/mL) of CEO and OEO were administered. qRT-PCR was used to measure gene expression levels of IL-1β and IL-8 from CMCs, occludin, ZO-1, and MUC2 from IECs, and Pax7 and MyoG from QMCs and EMCs. Cytotoxic effects of CEO and OEO were determined using an MTT assay; CEO and OEO did not show cytotoxicity at concentrations below 0.1 mg/mL in CMCs, IECs, QMCs, and EMCs. CEO reduced (P < 0.05) the LPS-induced increase of IL-1β and IL-8 in CMCs and increased (P < 0.05) ZO-1 and MUC2 in IECs. OEO suppressed (P < 0.05) the release of IL-8, increased ZO-1, and Pax7. Both CEO and OEO demonstrated microbicidal activity against sporozoite of E. tenella and C. perfringens bacteria, but only at doses 10-100 × higher than those that would be used in feed. These findings support our previous findings on other phytochemicals; both CEO and OEO are promising candidates for improved resilience in chickens not due to their direct antimicrobial effects, but due to gut physiological responses that take place at the level of the host.

The Potential Roles of IL-1β, IL-6, and RIPK3 in the Pathogenesis of Stevens–Johnson Syndrome/Toxic Epidermal Necrolysis

Background/Objectives: Stevens–Johnson Syndrome and Toxic Epidermal Necrolysis (SJS/TEN) are rare but severe skin conditions, often triggered by medications, that can be life-threatening. These conditions frequently affect the eyes, causing ocular surface disease, which can result in visual impairment or blindness. Although the exact mechanisms behind SJS/TEN remain unclear, key inflammatory mediators such as IL-1β, IL-6, and RIPK3 are believed to play critical roles in inflammation, necroptosis, and regulatory processes. Investigating these factors offers new insights into the disease’s underlying mechanisms and potential targets for treatment. This study aims to determine the roles of IL-1β, IL-6, and RIPK3 in the pathogenesis of SJS/TEN. Methods: The study examined the expression levels of IL-1β, IL-6, and RIPK3 in skin biopsies from patients with biopsy-confirmed SJS/TEN, using lichen planus as a positive control and normal skin as a baseline control. Immunohistochemistry was employed for this analysis. Additionally, the impact of SJS/TEN patient plasma on mitochondrial function was assessed in platelets and human corneal epithelial (H-CET) cells. Using a fluorescent plate reader, mitochondrial activity and superoxide ion levels were measured, comparing plasma from SJS/TEN patients to normal human plasma. Results: Skin biopsies from SJS/TEN patients showed a significantly higher expression of IL-1β, IL-6, and RIPK3 compared to both lichen planus and normal controls. Furthermore, plasma from SJS/TEN patients significantly reduced platelet viability and increased mitochondrial and total cellular superoxide ions, as demonstrated by elevated levels of MitoSOX Red and CellROX Red. Conclusions: These findings suggest that IL-1β, IL-6, and RIPK3 may contribute to the pathogenesis of SJS/TEN and highlight their potential as targets for therapeutic intervention.

P0183 GSDMD regulates interactions between macrophages and intestinal epithelial cells in inflammatory bowel disease

Abstract Background Our previous studies have demonstrated the significant role of the GSDMD-mediated pyroptotic pathway in inflammatory bowel disease (IBD). However, there remains some controversy surrounding the research findings concerning GSDMD in IBD. Methods Employing lentiviral transduction techniques, we manipulated GSDMD expression levels in THP-1 and HT-29 cells, assessing the impact of GSDMD on macrophage M1 polarization and epithelial barrier function. Further, co-culturing transduced THP-1 and HT-29 cells enabled the examination of changes in the intestinal barrier. Moreover, we generated conditional knockout mice with Gsdmdflox/floxLyz2-Cre and Gsdmdflox/floxVillin1-Cre, inducing experimental colitis with dextran sodium sulfate (DSS) to evaluate mucosal macrophage phenotypes and intestinal barrier integrity. Additionally, primary macrophages and intestinal epithelial cells were co-cultured, and downstream inflammatory factors were screened through transcriptome sequencing. Results Knockdown of GSDMD in THP-1 cells significantly reduces CD86 expression, indicating a decrease in M1 polarization efficiency, whereas overexpression of GSDMD yields opposite results. In HT-29 cells, overexpression of GSDMD followed by LPS treatment leads to a significant decrease expression of Claudin-1, Occludin, and Zo-1, whereas knockdown of GSDMD shows the opposite outcome. ELISA results demonstrate a correlation between GSDMD expression levels and levels of LDH, IL-1β, IL-18, TNF-α, and IL-6. In the inflammatory co-culture model of THP-1 and HT-29 cells with double knockdown of GSDMD, the severity of intestinal barrier damage and release of inflammatory factors is minimal. Conversely, in the inflammatory co-culture model of THP-1 and HT-29 cells with double overexpression of GSDMD, the intestinal barrier damage and release of inflammatory factors are maximal. Following DSS-induced colitis, compared to the Gsdmdflox/flox group, two conditional knockout mice exhibit reduced inflammatory evaluation. Furthermore, the protein expression of Zo-1, Occludin, and Claudin-1 in two conditional knockout mice is significantly higher than that in the Gsdmdflox/flox group. Flow cytometry results show that the proportion of M1 macrophages in the mucosal lamina propria is significantly lower in two conditional knockout mice compared to the Gsdmdflox/flox group. Additionally, primary macrophage and intestinal epithelial cell co-culture experiments, coupled with transcriptomic sequencing, revealed IL-1β as a downstream mediator of GSDMD-mediated macrophage-epithelial cell interplay. Conclusion It was confirmed that GSDMD serves as a key target in the pathological interplay between macrophages and epithelial cells.

Selective Gamma-Secretase Inhibition by CHF5074 Attenuates Inflammation and Neovascularization in a Murine Model of Choroidal Neovascularization

Purpose Chronic inflammation plays an important role in the pathogenesis of choroidal neovascularization (CNV). This study aimed to investigate the effect of the CHF5074, a γ-secretase inhibitor, on angiogenesis in a laser-induced CNV model and elucidate its possible molecular mechanism. Methods Male C57/BL6J mice aged between 6 to 8 weeks were employed to set up a laser-induced model of CNV. Then, CHF5074 was injected intraperitoneally on the day after laser modeling, as well as on the second, third, and fourth days. Immunofluorescence staining was used to evaluate the retinal and choroidal complex. The markers used were CD31 for neovascularization and IBA1 for microglia staining in ocular tissue slices. Fundus fluorescein angiography on days 3d, 7d, and 14d analyzed neovascularization and leakage areas. Inflammatory indicators were examined by Western blot (WB) and enzyme-linked immunosorbent assay (ELISA). High-throughput whole-tissue sequencing of retinal choroids identified relevant cell pathways. Key regulatory factors modulated by CHF5074 were identified via WB. Co-culture of BV2 cells and human umbilical vein endothelial cells (HUVEC) were used to explore the function of CHF5074 on the inhibition of tube formation. Results CHF5074 significantly decreased CD31 expression in the choroid on 3d, 7d, and 14d post-laser modeling (p < 0.05) and decreased both neovascularization and leakage areas (p < 0.05). Additionally, CHF5074 significantly lowered TNF-α, IL-10, and IL-1β expression levels in the choroid (p < 0.05), as demonstrated by WB analysis and ELISA. High-throughput whole-tissue sequencing identified P38-MAPK, JNK, and Wnt signaling pathways associated with neovascularization. CHF5074 decreased P38 protein phosphorylation (p < 0.05) as confirmed by WB analysis. CHF5074 inhibited the tube formation of HUVECs co-cultured with LPS and ATP-treated BV2 cells. Conclusion CHF5074 significantly suppresses angiogenesis in laser-induced choroidal neovascularization models, suggesting its potential as a novel agent for preventing and treating CNV.

Resveratrol alleviates blast lung injury by modulating the epithelial sodium channel (ENaC) via the PI3K/AKT pathway.

Blast lung injury (BLI) is a major cause of death in blast injuries, largely due to pulmonary edema. Effective clearance of alveolar fluid is critical for resolving pulmonary edema, with the epithelial sodium channel (ENaC) playing a key role in this process. Resveratrol (RES), a natural compound with known antioxidant and anti-inflammatory properties, has shown promise in treating respiratory diseases. However, its potential protective effects against BLI, particularly through ENaC modulation, remain unclear. In this study, complementary in vivo and in vitro models were used to investigate mechanisms underlying pulmonary edema following a gas explosion. We discovered that RES significantly alleviated BLI by decreasing pulmonary edema, pulmonary index, W/D ratio, oxidative stress, inflammatory cell infiltration, and TNF-α and IL-1β expression levels in rat lung tissue. RES upregulated the expression of ENaC and PI3K/AKT both in rats and in A549 cells. The knockout of the PI3K-p85 gene suppressed RES-induced upregulation of p-AKT and ENaC, reversing protein expression in A549 cells. Altogether, RES shows potential to attenuate blast-induced lung injury by upregulating ENaC, partly through the PI3K/AKT signaling pathway. This study highlights RES as a potential therapeutic agent for BLI and offers new insights into its mechanisms of action.

Zebrafish as a Visible Neuroinflammation Model for Evaluating the Anti-Inflammation Effect of Curcumin-Loaded Ferritin Nanoparticles.

It is crucial to inhibit the neuroinflammation response as it is a prominent factor contributing to the pathogenesis of neurodegenerative disorders. However, the limited development of neuroinflammation models dramatically hinders the efficiency of nanomedicine discovery. In recent years, the optically transparent zebrafish model provided unique advantages for in vivo imaging of the whole body, allowing the progression of the disease to be visualized. In this study, a lipopolysaccharide (LPS)-mediated zebrafish neuroinflammation model was established to visualize the brain distribution and quickly evaluate the anti-inflammation effect of human ferritin-loaded curcumin (Cur@HFn) nanoparticles. The Cur@HFn drug delivery system was successfully prepared and characterized. The HFn nanocage demonstrated significant brain accumulation and prolonged circulation in a zebrafish larval model. In the LPS-induced zebrafish model, Cur@HFn significantly reduced neutrophil recruitment within the brain region of the LPS-treated zebrafish. Additionally, Cur@HFn mitigated nitric oxide (NO) release and downregulated the mRNA expression levels of proinflammatory cytokines, including TNF-α and IL-1β. Lastly, Cur@HFn significantly reduced the damage of raphe nucleus neurons and alleviated the locomotion deficiency caused by LPS. Overall, our findings highlight that Cur@HFn is a promising drug delivery system for the targeted treatment of brain disorders. This zebrafish neuroinflammation model could be used for high-throughput in vivo drug screening and discovery.

Zeolite Imidazole Framework-8 Exacerbates Astrocyte Activation and Oxidative Stress in the Brain of Rats.

Metal-organic frameworks (MOFs) have been gaining significant attention due to their potential application in medicine. Here, we investigated the effect of zeolite imidazole framework-8 (ZIF-8) on neuro-behavioral parameters, histopathology, inflammation, and oxidative stress levels of rats' brain samples. Forty-eight male Wistar rats were injected by four injections of saline or ZIF-8 at different doses of 5, 10, or 20 mg/kg via the caudal vein. Y-Maze, Morris-Water Maze (MWM), and three chamber tests were conducted to explore working memory, spatial learning and memory, and social interactions, respectively. Histological staining and immunohistochemistry were used to evaluate pathological changes and astrocyte activation levels. The inflammation levels were measured using quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR). The total antioxidant capacity (TAC) and oxidative stress production were assessed by biochemical assays. The results showed that ZIF-8 induces neuromotor impairment dose-dependently. Although histopathological studies indicated increased neuronal loss, inflammatory changes, and elevated active astrocytes in the hippocampus, the expression levels of IL-1β and TNF-α were not significantly increased in ZIF-8-treated rats. The TAC level significantly reduced and the malondialdehyde (MDA) level remarkably increased in the brain tissues. Our findings suggest that administration of ZIF-8 induce neuromotor impairment, probably through amplified inflammation and oxidative stress.

New insight into the role of the pathway NLRP1 and NLRP3 inflammasomes and IL-33 in ultraviolet-induced cutaneous carcinogenesis.

Inflammasomes NLRP1 (NLR family pyrin domain containing 1) and NLRP3 are pivotal regulators of the innate immune response, activated by a spectrum of endogenous and exogenous stressors, including ultraviolet radiation (UVR). The precise molecular mechanisms underlying the activation of these inflammasomes remain unclear. Furthermore, the involvement of interleukin-33 (IL-33) in UVR-induced skin carcinogenesis is not well defined. The objective of this study is to evaluate the expression of interleukin genes (IL-33, IL-18, IL-1β) following the activation and silencing of NLRP1 and NLRP3 at various wavelengths and doses of UV radiation, and to correlate these expressions with pertinent tumor markers (e.g., Gli1, Gli2, FOXO3A, SerpinA1, SerpinA3, and EphB2). Cultures of keratinocyte cell lines were exposed to varying doses of UV radiation using specific lamps. To inhibit the expression of NLRP1 and NLRP3 genes, cells were transfected with targeted siRNAs. Gene expression of inflammasome components and effector proteins was quantified using Real-time PCR and ELISA. There was a marked upregulation in the expression levels of cytokine genes IL-18, IL-1β, and IL-33 upon exposure to UVB and UVA radiation, compared to non-irradiated keratinocytes. Silencing NLRP1 or NLRP3 via RNA interference in primary human keratinocytes resulted in a significant reduction of cytokine gene expression. Additionally, a notable increase in tumor marker gene expression was observed in cells with functional NLRP1 and NLRP3 following UV radiation, whereas silencing these inflammasome genes altered the expression profiles of these markers. This study provides a pioneering comprehensive assessment of the roles of NLRP1, NLRP3, and IL-33 in the pathogenesis of UV-induced cutaneous carcinogenesis. Our findings substantiate the role of IL-33 as a critical early danger signal elicited in response to inflammatory UV radiation, presumably regulated by inflammasomes.

Sea Anemone Kunitz Peptide HCIQ2c1 Reduces Histamine-, Lipopolysaccharide-, and Carrageenan-Induced Inflammation via the Suppression of Pro-Inflammatory Mediators

Inflammation is a physiological response of the immune system to infectious agents or tissue injury, which involves a cascade of vascular and cellular events and the activation of biochemical pathways depending on the type of harmful agent and the stimulus generated. The Kunitz peptide HCIQ2c1 of sea anemone Heteractis magnifica is a strong protease inhibitor and exhibits neuroprotective and analgesic activities. In this study, we investigated the anti-inflammatory potential of HCIQ2c1 in histamine- and lipopolysaccharide (LPS)-activated RAW 264.7 macrophages as well as in LPS-induced systemic inflammation and carrageenan-induced paw edema models in CD-1 mice. We found that 10 μM HCIQ2c1 dramatically decreases histamine-induced intracellular Ca2+ release and LPS-induced reactive oxygen species (ROS) production in RAW 264.7 macrophages. Moreover, HCIQ2c1 significantly inhibited the production of LPS-induced tumor necrosis factor α (TNF-α), inducible NO-synthase (iNOS), and 5-lipoxygenase (5-LO) but slightly influenced the IL-1β and cyclooxygenase-2 (COX-2) expression level in macrophages. Furthermore, intravenous administration by HCIQ2c1 at 0.1 mg/kg dose reduced LPS-induced TNF-α, IL-1β, COX-2, and iNOS gene expression in CD-1 mice. The subplantar administration of HCIQ2c1 at 0.1 mg/kg dose to mice significantly reduced carrageenan-induced paw edema by a factor of two, which is comparable to the effect of diclofenac at 1 mg/kg dose. Thus, peptide HCIQ2c1 has a strong anti-inflammatory potential by the attenuation of systemic and local inflammatory effects through the inhibition of intracellular Ca2+ release, the production of ROS and pro-inflammatory cytokines, and enzymes involved in arachidonic acid metabolism.

Construction with recombinant epitope-expressing baculovirus enhances protective effects of inactivated H9N2 vaccine against heterologous virus.

Although the use of inactivated vaccines has kept avian influenza (AI) outbreaks largely under control, they fail to prevent virus shedding. To enhance the efficacy of inactivated H9N2 AIV vaccines (InV), we constructed a multi-epitope recombinant baculovirus (BV-BNT) containing two B cell epitopes and nine T cell epitopes of H9N2 AIV for combined immunization with InV. The results showed that HI titer, IgG and IgM levels, and the percentage of B cells, CD4+ T cells, CD8+ T cells, and CD4+CD8+ T cells were significantly higher in the InV+BV-BNT immunization group than the InV immunization group. Besides, the expression levels of IL-1β, IFN-γ, IFN-α, IL-4, IL-13, and CXCLi1 were significantly higher in the InV+BV-BNT group than the InV group. Moreover, four conservative peptides (NP182-190, NP455-463, NS198-106, and NP380-393) significantly stimulated splenocytes to express IFN-γ in the InV+BV-BNT group instead of InV group. After heterologous virus challenging, the percentages of CD4+ T and CD8+ T cells were significantly upregulated in the InV+BV-BNT group compared to Inv group at 3 DPI. Viral loads in oropharyngeal of the InV+BV-BNT group was significantly lower than that in the InV group at 3 days post-infection (DPI). Furthermore, compared to the InV group, the virus positivity rate of oropharyngeal and cloacal swabs in the InV+BV-BNT group was lower at 5 DPI, with none positive at 7 DPI. Hence, this study indicated that the combined immunization of InV and BV-BNT could induce stronger humoral and cellular immune responses, shorten the detoxification period and reduce viral load compared to Inv alone, which suggests BV-BNT could act as a supplementary vaccine to potentially address the protection deficiency of the H9N2 inactivated vaccine.

Combination of inorganic nitrate and vitamin C prevents collagen-induced arthritis in rats by inhibiting pyroptosis.

Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disease characterized by synovial inflammation, cartilage damage, and bone loss. Although effective treatments are currently lacking, early interventions hold promise for alleviating RA symptoms. Inorganic nitrates and vitamin C (VC) are essential bioactive substances abundant in fruits and vegetables. Notably, nitrates and VC exhibit synergistic effects in a series of physiological and pathological conditions. In this study, we aim to examine the combination of nitrate and VC for preventing RA in a collagen-induced arthritis (CIA) rat model. Nitrate partly reduced foot swelling and arthritis scores and was more effective when combined with VC. Histopathological and radiological analyses revealed that nitrate + VC treatment alleviated synovial hyperplasia and bone loss. Additionally, nitrate + VC lowered the levels of TNF-α and IL-1β in serum as well as synovial tissue, decreased the expression of NF-κB and reduced the number of macrophages in synovial tissue. Compared to the CIA group, nitrate + VC decreased the levels of NLRP3 and GSDMD in macrophages, thus inhibiting pyroptosis. According to in vitro experiments, nitrate inhibited the activation of the NLRP3/caspase-1/GSDMD pathway in macrophages by conversion into nitrite. VC reduced the expression and phosphorylation of NF-κB in macrophages and thus reduced the expression levels of TNF-α and IL-1β. Therefore, nitrate/nitrite and VC may exert synergistic effects by blocking the interaction between NF-κB and NLRP3, further alleviating the inflammation and pyroptosis of macrophages, which provides a new strategy for RA prevention.