Expression Of Inflammatory Factors Research Articles

Fucoidan ameliorates lipid accumulation, oxidative stress, and NF-κB-mediated inflammation by regulating the PI3K/AKT/Nrf2 signaling pathway in a free fatty acid-induced NAFLD spheroid model

BackgroundNon-alcoholic fatty liver disease (NAFLD) is the most prevalent chronic liver disease worldwide. Previous studies have reported that fucoidan can relieve obesity and hepatic steatosis in vivo, although the molecular mechanism remains unclear. This study aimed to explore the effect and potential molecular mechanism of fucoidan in NAFLD using the free fatty acid (FFA)-induced NAFLD spheroid model.Materials and methodsThe spheroids were constructed by fusing the HepG2 and LX-2 cells. Spheroids and HepG2 cells were stimulated with FFAs and fucoidan, then the intracellular lipid contents and the oxidative stress levels (ROS/MDA/GSH/GR/GPx/NQO1/GCLC/HO-1) were detected. Furthermore, the regulation of PI3K/AKT/Nrf2 pathway and the expression of inflammatory factors (TNF-α and IL-6) were measured.ResultsFucoidan markedly reduced FFA-induced intracellular lipid accumulation in spheroids and HepG2 cells. Notably, fucoidan relieved FFA-induced oxidative stress by reducing the levels of ROS and MDA, and elevating the levels of GSH, GR, and GPx. Furthermore, fucoidan reduced FFA-induced oxidative stress by activating the PI3K/AKT/Nrf2 signaling pathway and by inhibiting ROS-induced P65 NF-κB activation and inflammatory responses via Nrf2 pathway activation.ConclusionsOur results demonstrated that fucoidan ameliorated FFA-induced lipid accumulation, oxidative stress, and NF-κB-mediated inflammation through the PI3K/AKT/Nrf2 signaling pathway in the spheroid and HepG2 cells model of NAFLD. These results provided new evidence for the clinical use of fucoidan in the treatment of NAFLD and its potential molecular mechanism of action.

Increased macrophage migration inhibitory factor is associated with inflammation in patients with rheumatoid arthritis.

The macrophage migration inhibitory factor (MIF) in the plasma, hydrops articuli, and synovium, and its relationship with laboratory indexes in patients with rheumatoid arthritis (RA) were determined, for the purpose to reveal the role of MIF on the pathogenesis of RA. MIF mRNA expression in PBMCs was detected by qPCR. Plasma MIF was measured by enzyme linked immunosorbent assay (ELISA). MIF in hydrops articuli and synovium from RA patientsand OA patients was evaluated by immunofluorescence (IF) and immunohistochemistry (IHC). The relationship between MIF and laboratory indexes of RA patients was analyzed. Human fibroblast-like synoviocytes (FLS) were treated with recombinant human MIF, and expression of inflammatory factors was determined by qPCR. The matrix metalloproteinase (MMP) 9 and extracellular regulated protein kinases (ERK)1/2 in FLS with MIF treatment were detected. MIF is significantly increased in plasma and hydrops articuli in RA patients. The expression of multiple inflammatory factors and MMPs was increased in RA patients and in FLS with rhMIF treatment. MIF was correlated with laboratory indexes in RA patients. Mechanistically, MIF promoted production of MMP9 by FLS through the ERK1/2 pathway. Our results indicated that increased MIF was correlated with disease activity of RA patients. These findings also suggested that MIF induced multiple inflammatory factors and MMP 9 in FLS via ERK 1/2 pathway. Key Points • MIF plays a key role in the initiation of RA by promoting the expression of various inflammatory factors in FLS and MMPs. • This study provides a basis for MIF-targeted RA clinical therapy and for exploring the feasibility of MIF as a therapeutic target for RA. • Increased MIF correlates with disease activity in RA patients.

Micheliolide attenuates sepsis-induced acute lung injury by suppressing mitochondrial oxidative stress and PFKFB3-driven glycolysis

BackgroundSepsis is a potentially fatal condition with a significant risk of death. Acute lung injury (ALI) is a life-threatening complication of sepsis, and the inflammatory response plays a critical role in sepsis-induced ALI. The protective effects of micheliolide (MCL) against renal fibrosis and leukemia have been demonstrated, but the precise underlying mechanisms remain unclear.MethodsIn vitro, lipopolysaccharides (LPS) and interferon-gamma (IFN-γ) were used to stimulate RAW264.7 cells and bone marrow-derived macrophages (BMDMs) to investigate the protective effect of MCL on sepsis-induced ALI. Cecal ligation and puncture (CLP) models were constructed in mice to induce ALI in vivo. The expression of inflammatory factors, macrophage polarization markers, and the glycolysis-related enzyme 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) were measured in vivo. Mitochondrial function, oxidative stress, and mitochondrial-related proteins were evaluated in vitro.ResultsMCL inhibited CLP-induced ALI, as evidenced by improvements in proinflammatory factor levels, lung wet/dry ratios, and histopathological findings. In vitro, MCL treatment significantly suppressed LPS + IFN-γ-induced M1-type polarization of RAW264.7 cells and BMDMs, as well as the production of inflammatory factors and oxidative stress. Mechanistic experiments revealed that MCL suppresses PFKFB3-driven glycolysis to reduce inflammation and activates the mitochondrial unfolded protein response (UPRmt) to alleviate mitochondrial stress. However, the therapeutic effect of MCL was diminished when PFKFB3 was overexpressed in cells.ConclusionThis study is the first to demonstrate that MCL attenuates sepsis-induced ALI by reducing M1-type macrophage polarization. Its therapeutic effect is closely related to the suppression of oxidative stress and PFKFB3-driven glycolysis.

Treatment of Acute Ulcerative Colitis with Zinc Hyaluronate in Mice.

Ulcerative colitis (UC) is a type of inflammatory bowel disease arising from numerous factors, while UC patients face insufficient treatment options and a high incidence of adverse reactions to the current therapies. As a functional food additive, hyaluronic acid plays a certain role in intestinal repair. In this study, we constructed a mouse model of dextran sulfate sodium (DSS)-induced UC to examine the effects and underlying mechanisms of action of zinc hyaluronate (ZnHA) on the pathogenesis of UC. ZnHA effectively alleviated key clinical UC symptoms, such as weight loss, loose stools, and bloody stools. Mechanistically, ZnHA attenuated the expression of inflammatory factors, such as tumor necrosis factor-α, interleukin (IL)-6, and myeloperoxidase while upregulating the expression of IL-10. Furthermore, through intestinal flora and short-chain fatty acid analyses, ZnHA was found to promote propionic acid production by enriching beneficial bacteria. ZnHA simultaneously enhanced the expression of tight junction proteins, specifically ZO-1 and occludin, thereby restoring intestinal barrier function. Overall, our findings elucidate the therapeutic potential of ZnHA in treating acute UC by inhibiting intestinal inflammation and regulating flora, while also providing further theoretical support for development of hyaluronic acid to treat this disease.

Pulsed electromagnetic field ameliorates the progression of osteoarthritis via the Sirt1/NF-κB pathway

BackgroundPulsed electromagnetic field (PEMF) is a non-invasive treatment that utilizes electromagnetic fields to reduce inflammation and promote tissue repair. However, PEMFs’ anti-inflammatory effect on osteoarthritis (OA) and the potential mechanism has not been fully elucidated.MethodsHuman chondrocytes (C28/I2) were stimulated with interleukin (IL)-1β with or without the treatment of PEMF. CCK-8 assay Kit was used to detect cell viability. RT-qPCR, ELISA, immunofluorescent staining and western blot was used to analyze relative markers of inflammatory response and extracellular matrix (ECM) under the treatment of PEMF and related mechanism. Besides, the significance role of Sirt1 was assessed by using the Sirt1 inhibitor (EX-527). Moreover, immunohistochemistry and immunofluorescence staining were carried out to evaluate the curative effect of PEMF on OA mice induced by the destabilization of the medial meniscus (DMM).ResultsPEMF inhibited IL-1β-mediated the expression of pro-inflammatory factors. Besides, PEMF alleviated IL-1β-induced degradation of ECM by increasing the expression of Col2a1 and ACAN, while inhibiting the expression of MMP13 and ADAMTS5. At the mechanism level, PEMF increased the expression of Sirt1 and inhibited IL-1β-induced the activation of NF-κB pathway. Furthermore, blocking Sirt1 with EX-527 attenuated the effect of PEMF on the inhibition of NF-κB pathway and the expression of ECM in IL-1β-induced chondrocytes. In vivo, PEMF-treated OA mice showed low modified mankin scores, reduced the number of osteophytes and preserved joint structure.ConclusionsOur results suggest that PEMF inhibits NF-κB pathway and blocks the expression of inflammatory factors by activating the expression of Sirt1, which may be a novel strategy for OA.

Study on the effect and mechanism of ZeXie decoction in treating MSU-induced acute gouty arthritis model through PI3K-AKT-mTOR signaling pathway.

The incidence of acute gouty arthritis (AGA) is annually increasing, significantly detrimenting the quality of life for patients. ZeXie decoction (ZXT), composed of Atractylodes macrocephala Koidz and Alisma rhizome (Sam.), a timeless formula detailed in "Synopsis of the Golden Chamber" of Chinese medical sage Zhong-Jing Zhang, has shown promising clinical application in treating AGA. Alisol B, a principal component of ZXT, remains however, elusive in its mechanism of action against AGA. This study aimed to delve into the anti-inflammatory effects of Alisol B, a key component within ZXT, and to clarify its mechanism of action in the treatment of AGA. We adopted a network pharmacology approach to pinpoint the core targets and pathways involved in ZXT and Alisol B's treatment of AGA patients. Molecular docking was conducted using Autodock software to investigate potential interactions between Alisol B and its target proteins. An in vitro inflammation model was subsequently established. The impact of Alisol B on the expression of inflammatory factors in BMDMs treated with MSU was evaluated using RT-qPCR, supplemented by comparison with the PI3K agonist 740 Y-P (740YPDGFR) treated BMDMs. Subsequently, the expression of EGFR, PIK3CA, PIK3CB, and JAK2 - key players in the PI3K/AKT/mTOR signaling pathway - was assessed via RT-qPCR and Western blotting. Finally, the effect of MSU treatment and Alisol B's treatment on macrophage polarization was determined by flow cytometry. Findings from network pharmacology and molecular docking suggest that Alisol B may modulate the PI3K-AKT-mTOR signaling pathway to treat AGA. In vitro experiments revealed that Alisol B inhibited the expression of inflammatory vesicles and pro-inflammatory factors by suppressing MSU-induced activation of the PI3K/AKT/mTOR signaling pathway. Additionally, Alisol B improved the cellular inflammatory environment, fostering the production of M2 cells, which could potentially repair cells within the inflammatory environment. Our research unveils that Alisol B curtails the production of inflammatory vesicles and pro-inflammatory cytokines while enhancing the production of anti-inflammatory factors by targeting the PI3K-AKT-mTOR signaling pathway in BMDMs. This may elucidate the pivotal mechanism of Alisol B in the treatment of AGA.

Icariin Induces Chondrocyte Degeneration via Activation of the NF-κB Signalling Pathway and Reduces the Inflammation Factor Expression Induced by Lipopolysaccharide.

To investigate the effect of icariin on chondrocytes by activating the nuclear factor kappa-B (NF-κB) signalling pathway and the mechanism of icariin in inducing chondrocytes degeneration. Chondrocytes were cultured invitro and treated with icariin at concentrations of 0, 0.01, 0.1, 10 and 20 μm. Cell proliferation was evaluated by cell counting kit-8 (CCK8) method, apoptosis was detected by TUNEL method and the expression of NF-κB was detected by Western blotting (WB) and RT-PCR. The nuclear localization of NF-κB p65 protein was observed by immunofluorescence staining. Lipopolysaccharide (LPS) was used to establish the inflammatory model, and WB and RT-PCR were used to detect the expression levels of tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6). After treatment with icariin, the proliferation ability of chondrocytes was markedly enhanced, and the apoptosis rate (AR) was markedly decreased (p < 0.05). In addition, icariin could inhibit the nuclear translocation of NF-κB p65 protein, promote the gene expression of Collagen II (Col-II) and Aggrecan in chondrocytes, and reduce the expression of NF-κB. Moreover, icariin markedly reduced the expression of IL-6 and TNF-α in LPS-induced inflammation (p < 0.05). Icariin can enhance chondrocyte proliferation, promote phenotypic gene expression by inhibiting the NF-κB signal transduction pathways, thus suppressing the chondrocyte degeneration, and can reduce the inflammation factor expression induced by LPS.

Hydrogen sulfide modulates acute lung injury in sepsis by inhibiting NLRP3 inflammasome activation in mice

Objective: To investigate the effects and mechanisms of the hydrogen sulfide donor GYY4137 on acute lung injury in sepsis. Methods: An acute lung injury model was established using the method of cecal ligation and puncture(CLP). Mice received an intraperitoneal injection of GYY4137 (50 mg/kg) or saline 30 minutes after surgery. C57BL/6J mice were divided into four groups: the sham operation group (Sham), the sham operation with GYY4137 group (GYY4137), the sepsis group (CLP), and the sepsis with GYY4137 group (CLP+GYY4137). Respiratory parameters (minute ventilation volume and expiratory flow 50) were measured using a whole-body plethysmography system. Lung tissue was evaluated by hematoxylin and eosin (H&E) staining, and inflammatory cells were identified by immunofluorescence. mRNA expression of inflammatory factors (interleukin-1β, interleukin-6) and adhesion molecules (vascular endothelial cadherin, intercellular cell adhesion molecule-1, vascular cell adhesion molecule-1) were quantified by real-time quantitative polymerase chain reaction (RT-PCR). Levels of NLRP3, Pro-IL-1β, IL-1β, and the cyclic guanosine monophosphate synthase (cGAS)/stimulator of interferon genes (Sting)/NF-κB signaling proteins were analyzed by Western blotting. In vitro, murine pulmonary microvascular endothelial cells were cultured and exposed to lipopolysaccharide (LPS, 1 μg/ml) and GYY4137 (25 μmol/L) to simulate sepsis-induced damage and to study the mechanism of hydrogen sulfide action. Levels of inflammatory factors and adhesion molecules in cells were quantified by RT-PCR. Data were analyzed for normal distribution using Shapiro-Wilk test, then variance homogeneity by Brown-Forsythe test. P-value<0.05 was set as statistical significance for all analyses. Results: (1) MV and EF50 values were significantly lower in the CLP group [(36.32±3.91)ml/min and (1.43±0.26)ml/s, respectively] compared to the Sham group [(50.14±6.07)ml/min and (2.70±0.46)ml/s, respectively]. These parameters were higher in the CLP+GYY4137 group [(45.83±2.33)ml/min and (2.02±0.16)ml/s, respectively] compared to the CLP group (P<0.05). (2) Severe lung tissue damage, alveolar collapse, increased septal thickening and exudation were observed in the CLP group and significantly reduced in the CLP+GYY4137 group (P<0.05). (3) Infiltration of inflammatory cells and mRNA levels of inflammatory factors and adhesion molecules were increased in the CLP group compared to the Sham group (P<0.05), and decreased in the CLP+GYY4137 group compared to the CLP group (P<0.05). (4) Protein levels of NLRP3, Pro-IL-1β, IL-1β, cGAS, Sting, and NF-κB were higher in the CLP group compared to the Sham group (P<0.05) and were reduced in the CLP+GYY4137 group compared to the CLP group (P<0.05). (5) LPS induced higher levels of inflammatory factors and adhesion molecules in pulmonary endothelial cells compared to the control, which were reduced in cells co-treated with GYY4137 and LPS (P<0.05). Conclusion: The hydrogen sulfide donor GYY4137 effectively modulates acute lung injury in septic mice by inhibiting NLRP3 inflammasome activation via the cGAS/Sting/NF-κB signaling pathway.

Qingfei Jiedu Huatan Formula Inhibits NLRP3 Inflammasome Activation to Attenuates Inflammation and Pyroptosis in Severe Pneumonia: Integrating Experimental Verification, Network Pharmacology and Transcriptomics.

Severe pneumonia (SP) represents an acute, critical condition characterized by high morbidity and mortality rates, along with numerous complications. The Qingfei Jiedu Huatan Formula (QJHF), a traditional Chinese medicine (TCM) formulation, is indicated for the treatment of severe pneumonia. However, its underlying therapeutic mechanisms remain uncertain. This study aimed to investigate the beneficial effects and molecular mechanisms of QJHF in the treatment of severe pneumonia. The anti-inflammatory properties of QJHF were assessed using a Klebsiella pneumoniae-induced rat model of SP and LPS-induced MH-S cells. Network pharmacology and transcriptomics were employed to identify potential targets and elucidate the molecular mechanisms underlying QJHF's action against SP. Pyroptosis in lung tissue and MH-S cells was examined via immunofluorescence and scanning electron microscopy. The expression of the NLRP3 inflammasome and its upstream regulatory pathways was measured through Western blot analysis. QJHF was found to alleviate pulmonary edema, enhance lung pathology, and improve the blood oxygenation index, while reducing inflammatory cell infiltration, expression of inflammatory factors, and lactic acidosis in SP rats. Serum containing QJHF-containing serum significantly reduced the secretion and transcription of inflammatory factors in MH-S cells. Network pharmacology and RNA-Seq analysis revealed potential targets modulated by QJHF against SP, showing a significant association between these targets and NLRP3 within the PPI network. Pathway enrichment analysis suggested that the NOD-like receptor, TNF, NF-κB, and JAK/STAT signaling pathways might regulate the NLRP3 inflammasome and coordinate the inflammatory response. Additionally, QJHF was shown to suppress NLRP3 inflammasome activation in rats with SP and in MH-S cells, which corresponded with markedly reduced levels of TNFR1, TRAF2, TAK1, phosphorylated p65, IκBα, JAK2, and STAT3. QJHF effectively attenuated the inflammatory response in severe pneumonia by inhibiting macrophage-mediated inflammation and NLRP3 inflammasome activation through TNF, NF-κB, and JAK/STAT signaling pathways.

Identify of blood glucose metabolism regulation pathways and related proteins in the db/db mouse model through iTRAQ quantitative mass spectrometry.

Diabetes cardiomyopathy (DCM) has become the main cause of death of diabetes patients due to heart failure. As the initial unclear symptoms and complex underlying pathological mechanisms, it presents significant challenges for early diagnosis. It is essential to explore valuable biomarkers to enhance our understanding involved in DCM. Twelve-week-old db/db model mice (diabetes group) and normal mice (control group) were maintained in a specific pathogen-free (SPF) environment. Body weight, blood glucose, and insulin levels were measured regularly. At 26 weeks, cardiac tissue was collected to assess oxidative stress, inflammatory factors, and fibrosis markers, followed by histopathological examination. Meanwhile, iTRAQ-based quantitative mass spectrometry was employed to identify differentially expressed proteins (DEPs) in cardiac tissue. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) methods were utilized to analyze their biological functions and signaling pathways. Furthermore, specific proteins and their associated signaling pathways related to blood glucose regulation were validated in cardiac tissue and myocardial cells using western blot and immunofluorescence analysis. The mice in the control group were active and exhibited healthy growth, whereas those in the diabetes group showed increased food and water intake. Furthermore, there were significant elevations in blood glucose concentration, insulin levels, and body weight in the diabetes group. Histopathological examinations revealed that the myocardium in the diabetes group was markedly hypertrophic due to persistent hyperglycemia, accompanied by muscle fiber disarray, nuclear damage, and a significant increase in the expression of oxidative stress and inflammatory factors. Mass spectrometry analysis identified a total of 107 DEPs, comprising 83 up-regulated and 24 down-regulated proteins. Notably, the most significant difference was observed in the regulation of the glycogen metabolism enzyme (PYGM). GO and KEGG analyses indicated that the DEPs were primarily involved in glycogen metabolism, catalysis, and oxidative stress, with signaling pathways related to fatty acid metabolism, the PPAR pathway, insulin resistance, and the tricarboxylic acid cycle. Subsequent immunofluorescence and western blot analysis confirmed that hyperglycemia inhibits the PI3K/AKT signaling pathway and upregulates the expression of PYGM. Conversely, the knockout of PYGM significantly enhanced the activity of the PI3K/AKT signaling pathway. The research suggests that the inhibition of PYGM enhances the activity of the PI3K/AKT signaling pathway in patients with DCM, and then help to reduce blood glucose levels. This molecular mechanism exerts a protective effect on DCM. These findings highlight the potential of targeting PYGM as a novel biomarker for the early diagnosis, which may serve as an effective therapeutic strategy.

Human cytomegalovirus tegument protein UL23 promotes gastric cancer immune evasion by facilitating PD-L1 transcription

Immune checkpoint therapy targeting PD-1/PD-L1 has shown promise in treating tumors, however, its clinical benefits are limited to a subset of gastric cancer (GC) patients. Recent research has highlighted a the correlation between PD-L1 expression and the clinical efficacy of anti-PD-1/PD-L1 therapies. Human cytomegalovirus (HCMV) has been implicated in GC, but its specific role in modulating this disease remains elusive. In this study, we analyzed clinical tissue samples using bioinformatics and real-time quantitative polymerase chain reaction (RT-qPCR). We found that GC tissues infected with HCMV presented higher PD-L1 expression compared to those without virus. Furthermore, we demonstrated that HCMV infection enhances PD-L1 expression in GC cells. Cytotoxicity assays revealed that HCMV modulates cancer immune responses via the PD-1/PD-L1 pathway. Mechanistically, we showed that HCMV activates the PI3K-Akt signaling cascade and modulates PD-L1 expression through its tegument protein UL23. Functionally, increased UL23 expression leads to elevated PD-L1 levels, which diminishes tumor cell sensitivity to T-cell-mediated cytotoxicity and triggers T-cell apoptosis. Additionally, in vivo experiments revealed that UL23-induced PD-L1 upregulation inhibits CD8+ T-cell infiltration and reduces the expression of inflammatory factors in tumor microenvironment, ultimately weakening antitumor immunity. Our findings reveal a novel mechanism whereby HCMV and its tegument protein UL23 contribute to cancer immunosuppression through the regulation of PD-L1 expression. This discovery may serve as a potential therapeutic target for enhancing the efficacy of cancer immunotherapy.

FBN2 promotes the proliferation, mineralization, and differentiation of osteoblasts to accelerate fracture healing

Fracture is a disease in which the continuity of bone is interrupted or the integrity of bone is destroyed due to various reasons. It can be life-threatening when severe fractures occur. The RNA-seq datasets related to ‘fracture’ were screened and the common differentially expressed genes (DEGs) were determined. Protein-protein interaction network was constructed to identify hub genes. The fracture mice model was constructed and HE staining was performed to observe the histological characteristics of fracture. The expression of inflammatory factors and hub genes were evaluated by ELISA and qRT-PCR. CCK-8 assay, flow cytometry and Alizarin Red S staining were performed to evaluate the effects of fibrillin2(FBN2) on viability, apoptosis and mineralization of MC3T3E1 cells, respectively. Western blot was executed to measure expression of osteogenic markers (ALP and RUNX2). A total of 78 common DEGs were screened from GSE157460 and GSE152677 datasets. FBN2 was down-regulated in fracture and identified as the hub gene. In fracture mice, the thickness of the compact bone decreased in Day 1, accompanied by callus and woven bone formation, filled with a large number of osteoblasts, while IL-1β, IL-6 and TNF-α levels were increased. FBN2 enhanced cell viability and mineralization, suppresses apoptosis of MC3T3E1 cells, and facilitated the expression of ALP and RUNX2. Meanwhile, the knockdown of FBN2 demonstrated opposing trends. Through bioinformatics analysis, FBN2 was identified as the hub gene in fracture, and FBN2 promoted the proliferation, mineralization, and differentiation of osteoblasts, thereby accelerating fracture healing.

Itaconate promotes mitophagy to inhibit neuronal ferroptosis after subarachnoid hemorrhage.

Subarachnoid hemorrhage (SAH), representing 5-10% of all stroke cases, is a cerebrovascular event associated with a high mortality rate and a challenging prognosis. The role of IRG1-regulated itaconate in bridging metabolism, inflammation, oxidative stress, and immune response is pivotal; however, its implications in the early brain injury following SAH remain elusive. The SAH nerve inflammation model was constructed by Hemin solution and BV2 cells. In vitro and in vivo SAH models were established by intravascular puncture and Hemin solution treatment of HT22 cells. To explore the relationship between IRG1 and neuroinflammation by interfering the expression of Irg1 in BV2 cells. By adding itaconate and its derivatives to explore the relationship between mitophagy and ferroptosis. IRG1 knockdown increased the expression of inflammatory factors and induced the transformation of microglia to pro-inflammatory phenotype after SAH; Itaconate and itaconate derivative 4-OI can reduce oxidative stress and lipid peroxidation level in neuron after SAH, and reduce EBI after SAH; IRG1/ itaconate promotes mitophagy through PINK1/Parkin signaling pathway to inhibit neuronal ferroptosis. IRG1 can improve nerve inflammation after SAH, M2 of microglia induced polarization. IRG1/ Itaconate participates in mitophagy through PINK1/Parkin to alleviate neuronal ferroptosis after SAH and play a neuroprotective role.

Effect of intestinal microbiota on adaptation to overcrowding stress in grouper (Epinephelus fuscoguttatus♀×E. lanceolatus♂).

Density is an important aquaculture parameter. When the pearl gentian grouper (Epinephelus fuscoguttatus♀×E. lanceolatus♂) is farmed intensively, it could lead to a degradation in genetic resources and an increase in disease outbreaks. The composition of the intestinal microbiota plays a key role in creating a specific intestinal microecosystem, which is essential for the survival, growth, and immune response of the host under environmental stress like overcrowding. This study utilized 16S rRNA sequencing and metabolomics analysis techniques to investigate the differences in intestinal microbial community stability of grouper under different stocking time and density pressure conditions. The research results showed that compared to the low-density group, the high-density group of groupers experienced an increase in mortality rate and feed coefficient in the early stages of culture, while the weight gain rate decreased. Differential analysis of intestinal microbial communities revealed significant differences in the gut microbiota of grouper between different density groups after 10 days of culture, but no significant differences were observed after 20 days of culture. At the same time, intestinal histopathology showed that the high-density group of groupers exhibited a reduction in intestinal villi length and thickness of the intestinal wall after 10 days of culture. However, the intergroup differences had reduced after 20 days of culture. Furthermore, high density cultivation upregulated the expression of inflammatory factors like IL-1β, TNF-α, IL-8, and IL-6 in the intestinal tract of groupers after 10 days of culture. However, after 20 days of culture, the expression levels of intestinal inflammatory factors in both the high-density and low-density groups of groupers were significantly reduced, and the differences between the intergroup diminished. Through correlation analysis of differential metabolites and species in the intestine, multiple metabolites significantly upregulated and associated with the upregulation of the Staphylococcus genus were identified in the intestinal tract of groupers after 20 days of high-density cultivation. The selected four associated metabolites (including creatine, fosinopril, 4-aminobutyric acid, and guanidinopropanoic acid) were validated to significantly reduce the expression of cellular inflammatory factors using the self-established grouper head kidney (HK) cell line. In conclusion, density pressure in the early culture period could affect the stability of the intestinal microbial environment of grouper. As aquaculture time increases, the intestinal microbial community of grouper drives the body's anti-inflammatory response and enhanced its adaptation to density pressure by regulating own structure and secretion of metabolites.

Cycloastragenol targets Fpr2 to inhibit the TLR4/NF-κB signaling pathway and alleviate neuroinflammation in Parkinson's disease.

Parkinson's disease (PD) is a neurodegenerative disease, and neuroinflammation is an important factor in its pathogenesis. Therefore, improving neuroinflammation has become a key direction in PD research. Cycloastragenol (CAG) is one of the active ingredients in Astragalus membranaceus, which has pharmacological activities such as anti-inflammatory, antioxidant, and neuroprotective effects. However, there are few reports on its pharmacological effects on PD. Therefore, it is necessary to comprehensively evaluate the pharmacological effects of CAG on PD and elucidate the potential mechanisms of action, providing new ideas for drug development in PD. To comprehensively and systematically evaluate the pharmacological effects of CAG on PD and reveal its potential mechanisms of action. Firstly, the pharmacological effects of CAG on cell viability, cytotoxicity, behavior, and pathology were evaluated using PD in vitro (MPP+ induced SH-SY5Y cells) and in vivo models (MPTP induced mouse model). Furthermore, potential targets and signaling pathways will be screened based on metabolomics and transcriptomics. Ultimately, the connection between the target and the signaling pathway will be validated to elucidate the potential mechanism by which CAG exerts its effects. CAG can significantly improve the behavioral indicators of PD mice, enhance neuronal vitality, and improve neuroinflammatory levels by inhibiting the expression of inflammatory factors. In addition, CAG can target and activate the expression of Fpr2, thereby regulating the TLR4/NF-κB signaling pathway and promoting the resolution of inflammation.

Phillyrin ameliorates DSS-induced colitis in mice via modulating the gut microbiota and inhibiting the NF-κB/MLCK pathway.

Phillyrin (PHY), also known as forsythin, is an active constituent isolated from the fruit of Forsythia suspensa (Thunb.) Vahl (Oleaceae). It exhibits anti-inflammatory, anti-viral, and antioxidant properties. However, the precise impact of PHY on colitis induced by dextran sodium sulfate (DSS) and its mechanism remain elusive. The present investigation revealed that PHY (12.5, 25.0, and 50.0 mg/kg) exhibited significant therapeutic efficacy in protecting mice against DSS-induced colitis. This effect was manifested as reduced weight loss, a shortened colon, increased secretion of inflammatory factors, increased intestinal permeability, and an enhanced disease activity index in mice with ulcerative colitis (UC). Molecular investigations have determined that PHY mitigates the nuclear translocation of nuclear factor kappa B, thereby downregulating myosin light-chain kinase-driven myosin light-chain phosphorylation. This mechanism results in the preservation of the integrity of the intestinal barrier. The outcomes of 16S rRNA sequencing suggest that PHY (50 mg/kg) augmented the relative abundance of certain probiotic strains, including Lactobacillaceae and Lachnospiraceae. Additionally, PHY supplementation elevated the short-chain fatty acid contents within the intestinal contents of mice with UC. In conclusion, pre-treatment with PHY may ameliorate the DSS-induced UC in mice by lowering the expression of inflammatory factors, protecting intestinal barrier function, and enhancing the structure of the intestinal flora.IMPORTANCEThe protective effect of phillyrin on DSS-induced colitis was explained for the first time, and the anti-inflammatory effect of phillyrin was demonstrated by fecal microbiota transplantation experiments mainly through intestinal flora.

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.

The Experimental Study of Double-Layer Heterogeneous CA Scaffold in Promoting the Surface Shape Recovery and Internal Osteogenesis of Alveolar Bone.

In this work, double-layer heterogeneous CA scaffolds were designed for alveolar bone defects. The outer layer featured high hardness and slow degradation, and large pores and rapid degradation characterized the inner layer. The CA scaffold morphology was akin to bone defects, and its direct implantation reduced the operation time. A higher concentration of CA resulted in smaller pores and slower degradation. CA can promote the formation of mineralized nodules and the expression of genes related to mineralization without inducing cytotoxic effects. It also promoted the expression of cellular inflammatory factors, potentially through the TLR4 pathway. In vivo studies confirmed that CA did not promote the aggregation of inflammatory cells or the expression of inflammatory factors. In conclusion, the scaffold's characteristics of high surface hardness and slow degradation were beneficial for surface osteogenesis and maintaining the defect's shape and osteogenic space. Conversely, rapid internal degradation favors the formation of bone tissue.

Effect of 0.05% cyclosporine A eye drops on the healing process of corneal epithelial defects in rats

0.05% cyclosporine A eye drops is a kind of new medication for dry eye after corneal refractive surgeries. However, it is still unclear how this eye drops affect the healing process of corneal epithelial defects caused by operative procedures. In this vivo study, the effect of 0.05% cyclosporine A eye drops on the healing process of the corneal epithelium was assessed in a rat model featuring mechanically induced central corneal epithelial defects. These Sprague-Dawley rats were randomly divided into three groups, the 0.05% cyclosporine A eye drops group (CsA group), the 0.1% sodium hyaluronate eye drops group (HA group), and the model control group (MC group). The epithelial healing, tear secretion, epithelial microvilli, expression of apoptosis markers (TUNEL, p53, and bcl-2 proteins), inflammatory factors (IL-1β, TNF-α, and IL-6), and epidermal growth factor (EGF) were detected in the current study using corneal fluorescein sodium staining, phenol red thread test, transmission electron microscopy (TEM), immunofluorescence, and enzyme-linked immunosorbent assay (ELISA), respectively. Compared to the other two groups, lower expression of apoptosis markers and inflammatory factors were exhibited in the CsA group, along with its faster and better epithelial healing and higher tear secretion (P<0.05). In conclusion, 0.05% cyclosporine A eye drops effectively promote the healing process for corneal epithelial defects in rats, potentially offering advantages for rapid corneal recovery after refractive surgical procedures.

Beclin1 regulates yak endometrial inflammation and TLR4/NF-κB signaling pathway through autophagy/non-autophagy function.

Beclin1 is an autophagy related factor, and it is capable of mediating non-autophagy functions, too. Yak endometritis represents a significant obstetric ailment that impedes the normal breeding process. The current understanding of the beclin1 effect on endometrial inflammation in yak remains limited. Accordingly, this study initially examined the expression profile of beclin1 in yak endometritis in vitro and vivo. Subsequently, the beclin1 was targeted inhibit through small interfering RNA (siRNA), with the objective of elucidating the regulatory function of beclin1 in yak endometritis. The findings reveal that expression of beclin1 in inflammatory tissues of yak endometrium is markedly elevate in comparison to control group, and predominant localization in the cytoplasm of the endometrium and uterine glands. 1µg/mL Lipopolysaccharide (LPS) was demonstrated to induce yak endometrial epithelial cells (YEECs) inflammation and increase the expression of beclin1. YEECs are disposed with 1μg/mL LPS, resulting in a gradual increase of p62 expression from 0h to 6h, and significant decrease at 12h, at 9h to 12h the expression of LC3 significant increase. These findings indicate that LPS impairs autophagy during the initial stages of inflammation, complete autophagy is occurred in cells during the subsequent phase. Initial stages of inflammation, inhibit beclin1 result significantly reduced expression of inflammatory factors (TNF-α and IL-1β) and TLR4/NF-κB signaling pathway (p65, IκBα phosphorylation, p65 nuclear translocation) compared to the control group. When complete autophagy occurred, inhibit beclin1 inhibit autophagy, result in a significantly higher expression of inflammatory factors (TNF-α and IL-1β) and TLR4/NF-κB signaling pathway than the control group. In conclusion, this study demonstrates for the beclin1 exerts both autophagic and non-autophagic functions during the inflammatory process in YEECs, making it become a potential target for the cure and diagnosis of various yak endometritis.