Toll-like Receptor Signaling Pathway Research Articles

Identification of Immune Infiltration-Associated CC Motif Chemokine Ligands as Biomarkers and Targets for Colorectal Cancer Prevention and Immunotherapy

Colorectal cancer (CRC) is the third most common cancer globally, with limited effective biomarkers and sensitive therapeutic targets. An increasing number of studies have highlighted the critical role of tumor microenvironment (TME) imbalances, particularly immune escape due to impaired chemokine-mediated trafficking, in tumorigenesis and progression. Notably, CC chemokines (CCLs) have been shown to either promote or inhibit angiogenesis, metastasis, and immune responses in tumors, thereby influencing cancer development and patient outcomes. However, the diagnostic and prognostic significance of CCLs in CRC remains unclear. In this study, multiple online tools for bioinformatics analyses were utilized. The findings revealed that the mRNA expression levels of CCL3, CCL4, and CCL26 were significantly elevated in CRC tissues compared to normal tissues, whereas CCL2, CCL5, CCL11, CCL21, and CCL28 mRNA levels were markedly downregulated. Additionally, dysregulation of CCL4, CCL5, and CCL21 was strongly associated with clinical staging, and elevated levels of CCL4, CCL11, and CCL28 were linked to significantly prolonged survival in CRC patients. Functional enrichment analysis indicated that the cellular roles of CCLs were predominantly associated with the chemokine, Wnt, and Toll-like receptor signaling pathways, as well as protein kinase activity. Furthermore, transcriptional regulation of most CCLs involved RELA and NFKB1. Key downstream targets included members of the SRC family of tyrosine kinases (HCK, LYN, and LCK), serine/threonine kinases (ATR and ATM), and others such as CSNK1G2, NEK2, and CDK2. Moreover, CCLs (CCL2, CCL3, CCL4, CCL5, CCL11, CCL21, and CCL28) exhibited strong correlations with major infiltration-related immune cells, including B cells, CD8+ T cells, CD4+ T cells, macrophages, neutrophils, and dendritic cells. In conclusion, our study provides novel insights into the potential utility of CCLs as biomarkers and therapeutic targets for CRC prevention and immunotherapy.

SFAs facilitates ceramide's de novo synthesis via TLR4 and intensifies hepatocyte lipotoxicity.

Non-alcoholic steatohepatitis (NASH), an advanced manifestation of non-alcoholic fatty liver disease (NAFLD), is characterized by hepatocyte injury, inflammation, and fibrosis. Saturated fatty acids (SFAs) have emerged as key contributors to hepatocyte lipotoxicity and disease progression. Toll-like receptor 4 (TLR4) acts as a sentinel for diverse ligands, including lipopolysaccharide (LPS) and endogenous molecules like palmitic acid (PA)-induced ceramide (CER) accumulation, promoting hepatocyte demise. However, the intricate mechanisms underlying TLR4's modulation of ceramide metabolism and their concerted effect on SFA-mediated hepatotoxicity remain elusive. A NASH mouse model with liver-specific TLR4 knockdown was established through palm oil feeding and AAV2/8 tail vein injection. Histological and biochemical assessments were conducted to evaluate the mice's condition and liver damage extent. Liquid chromatography-mass spectrometry (LC-MS) was employed to quantify ceramide levels in liver tissues, offering insights into NASH mechanisms. The PO-fed model exhibited elevated serum ALT, AST, and liver TG levels, enhancing lipid accumulation and hepatocellular damage. TLR4 knock-down reduced liver mass and the liver-to-body weight ratio, signifying a decreased hepatic burden. Histopathological evaluations revealed substantial improvement in hepatic steatosis in TLR4-silenced PO-fed mice, with diminished lipid droplets and inflammatory infiltrates. LC-MS analysis showed a marked decrease in long-chain ceramides (C14, C16, C20) in TLR4-knockdown PO-fed mice. Furthermore, expression of MyD88, SPTLC1, SPTLC2, and inflammatory markers IL-1β, IL-6, TNF-α were significantly attenuated. SFAs activate the TLR4 signaling pathway via MyD88, fostering ceramide de novo synthesis, which exacerbates hepatocyte lipotoxicity and accelerates NASH progression.

5-methoxytryptophan protects against toll-like receptor 2 mediated renal tissue inflammation and fibrosis in a murine unilateral ureteral obstruction model.

5-methoxytryptophan (5-MTP) is an anti-inflammatory metabolite. Several recent reports indicate that 5-MTP protects against post-injury tissue fibrosis. It was unclear how 5-MTP controls tissue fibrosis. We postulated that 5-MTP attenuates renal interstitial fibrosis by blocking toll-like receptor 2 (TLR2) and transforming growth factor β (TGFβ) signaling pathways. The effect of 5-MTP on renal fibrosis was evaluated by pretreatment of wild-type UUO mice with intraperitoneal administration of 5-MTP. To determine whether 5-MTP attenuates fibrosis by inhibiting TLR2 and TGFβ signaling pathways, we evaluated the effect of 5-MTP on TLR2-induced fibroblast phenotypic switch in NRK-49F fibroblasts and TLR2 and TGFβ signaling pathways in human proximal tubular epithelial cells (HPTEC) and RAW264.7 macrophages stimulated with Pam3CSK4 (Pam3) or TGFβ1. UUO-induced renal fibrosis was abrogated in tlr2-/- mice consistent with a crucial role of TLR2 in UUO-induced renal fibrosis. UUO-induced macrophage infiltration and pro-fibrotic cytokine production in renal tissues were suppressed by tlr2 knockout. 5-MTP attenuated renal tissue fibrosis accompanied by reduction of macrophage infiltration and IL-6 and TGFβ levels. 5-MTP inhibits TLR2 upregulation and blocks TLR2-MyD88-TRAF6 signaling pathway in macrophages. Furthermore, 5-MTP blocked Pam3- and TGFβ1-induced phenotypic switch of NRK-49F to myofibroblasts and inhibited Pam3- and TGFβ1-induced signaling pathways in HPTECs and RAW264.7 cells. 5-MTP effectively protects against UUO-induced renal interstitial fibrosis by blocking TLR2 and TGFβ signaling pathways.

Intracellular CIRP promotes liver regeneration via STAT3 signaling pathway activation after partial hepatectomy in mice.

Cold‑inducible RNA‑binding protein (CIRP) is a cold shock protein implicated in the regulation of multiple biological processes depending on its cellular localization. However, to the best of our knowledge, the role of CIRP in liver regeneration and injury after hepatectomy has not been investigated. The present study was therefore designed to explore whether CIRP is involved in liver regeneration after hepatectomy and its specific role and underlying molecular mechanism. The overall involvement of CIRP in liver regeneration and injury after hepatectomy was evaluated in CIRP‑deficient mice. C23, an antagonist of extracellular CIRP, was used to assess the effect of extracellular CIRP on liver regeneration and injury after hepatectomy. CIRP overexpression and short hairpin RNA plasmids were transfected into HepG2 cells to study the effect of intracellular CIRP on cell proliferation. The effects of extracellular CIRP on cell proliferation and injury were determined via the use of recombinant CIRP protein to stimulate HepG2 cells in vitro. The results indicated that both hepatic and serum CIRP levels significantly increased after partial hepatectomy. Additionally, CIRP deficiency impaired liver regeneration but alleviated liver injury after partial hepatectomy in mice. C23 administration attenuated liver injury and suppressed endoplasmic reticulum (ER) stress and oxidative stress. Loss‑ and gain‑of‑function analyses in HepG2 cells indicated that an increase in intracellular CIRP promoted cell proliferation via signal transducers and activation of transcription 3 (STAT3) signaling pathway activation. Moreover, recombinant CIRP had no effect on cell proliferation or STAT3 phosphorylation but induced ER stress, which was blocked by TAK242, an inhibitor of Toll‑like receptor 4 (TLR4), in HepG2 cells. Taken together, the results of the present study demonstrated that intracellular CIRP promotes liver regeneration by activating the STAT3 pathway, whereas extracellular CIRP induces ER stress possibly via the TLR4 signaling pathway after hepatectomy.

Polysaccharides of Atractylodes Macrocephala Koidz Alleviate LPS-Induced Bursa of Fabricius Injury in Goslings by Inhibiting EREG Expression.

The bursa of Fabricius (BF) plays crucial roles in the goslings' immune system. During waterfowl breeding, the presence of lipopolysaccharides (LPSs) in the environment can induce inflammatory damage in geese. Polysaccharides of Atractylodes macrocephala Koidz (PAMKs), as the main active component of the Chinese medicine Atractylodes macrocephala, have significant immune-enhancing effects. Accordingly, this study intended to investigate the effect of PAMKs on LPS-induced BF injury in goslings. Two hundred 1-day-old goslings (half male and half female) were selected and randomly divided into control, PAMK, LPS, and PAMK + LPS groups. The control and LPS groups were fed the basal diet, and the PAMK and PAMK + LPS groups were fed the basal diet containing PAMKs at 400 mg/kg. The goslings in the LPS and PAMK + LPS groups were injected intraperitoneally with LPS at a concentration of 2 mg/kg on days 24, 26, and 28 of this study. The control and PAMK groups were injected with equal amounts of saline. On the 28th day, 1 h after the LPS injection, the BF and serum were collected and analyzed for organ indices, cytokines, antioxidant indicators, and histological observations. Histological examination and HE staining demonstrated that the PAMK treatment ameliorated the LPS-induced BF atrophy, structural damage, increased cellular exudation, and reticulocyte hyperplasia in the goslings. The cytokine and antioxidant marker analyses in the BF cells demonstrated that the PAMK treatment mitigated the LPS-induced increase in the interleukin-1β (IL-1β), malondialdehyde (MDA), and inducible nitric oxide synthase (iNOS) levels, as well as the decrease in the transforming growth factor-β (TGF-β) and superoxide dismutase (SOD) activities. Further transcriptome sequencing identified a total of 373 differentially expressed genes (DEGs) between the LPS and PAMK + LPS groups. The KEGG enrichment pathway analysis showed that the DEGs were significantly enriched in the Toll-like receptor, p53, MAPK, GnRH, and ErbB signaling pathways. Among them, EREG played key roles in the activation of the MAPK, GnRH, and ErbB signaling pathways. Further research showed that the addition of PAMKs significantly inhibited the LPS-induced EREG expression, increased the cell viability, promoted the cell cycle entry into the S and G2 phases, and inhibited apoptosis. Meanwhile, PAMKs can reduce the protein expression of p-JNKs and c-FOS by inhibiting EREG. In summary, this study found that PAMKs could alleviate LPS-induced BF injury in goslings by inhibiting the expression of EREG.

Shenlian Decoction Ameliorates LPS-Related Inflammation in db/db Mice: Coupling Network Pharmacology With Experimental Verification.

Background: Shenlian (SL) decoction, a renowned traditional Chinese formula for diabetes mellitus, has also been employed to treat intestinal disorders. Previous studies have demonstrated the efficacy of SL decoction in regulating blood glucose and intestinal bacteria. Nevertheless, further analysis is required to elucidate the mechanistic link between SL decoction-mediated improvement of intestinal function and treatment of Type 2 diabetes mellitus (T2DM). Methods: Firstly, the active ingredients of SL decoction were sourced from the Traditional Chinese Medicine System Pharmacology (TCMSP) database, with putative targets of active ingredients being predicted using the same database. Secondly, the Online Mendelian Inheritance in Man (OMIM) and GeneCards databases were employed to screen the aforementioned targets that act on T2DM, and protein-protein interaction (PPI) networks were constructed in accordance with the results. Thirdly, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted using the Database for Annotation, Visualization, and Integrated Discovery (DAVID), which resulted in a comprehensive analysis of the association between SL decoction for the treatment of T2DM and the modulation of intestinal functions. Finally, the effect of the SL decoction on predicted lipopolysaccharide (LPS)-related targets, as well as intestinal function markers, was validated through in vivo experimentation. Results: A total of 36 active ingredients and 145 potential targets of SL decoction were predicted. GO enrichment analysis indicated that the principal biological processes by which the SL decoction acted against T2DM were responses to LPSs, while KEGG enrichment analysis identified the nuclear factor kappa B (NF-κB) signaling pathway and toll-like receptor signaling pathway as the key pathways involved. The in vivo experiments showed that SL decoction improved glycolipid metabolism indexes, inflammatory factor levels, and LPS levels in db/db mice. The immunohistochemical results demonstrated that the SL decoction restored the expression of Occludin, Claudin-1, and ZO-1 in the intestine and inhibited the expression of toll-like receptor 4 (TLR4), myeloid differentiation primary response gene 88 (MYD88), and NF-κB in both the intestine and pancreas. Furthermore, it may influence the levels of short-chain fatty acids (SCFAs) in feces. Conclusions: This research investigated the multigene pharmacological mechanism of SL decoction against T2DM using network pharmacology and in vivo experiments. SL decoction treatment of T2DM may reverse inflammation by inhibiting LPS-related pathway activation and improving intestinal function.

High glucose enhances malignant progression of MDA-MB-231 cells through cumulative effect

Previous investigations have shown that high glucose can promote breast cancer progression. However, the relationship between high glucose microenvironment and triple-negative breast cancer (TNBC) remains to be explored. In this study, we performed RNA-seq to explore the effect of short-term high glucose and long-term high glucose on MDA-MB-231 cell line. A total of 896 highly ranked differentially expressed genes (DEGs) were identified, including 57 DEGs of short-term high glucose group and 839 DEGs of long-term high glucose group. The DEGs of short-term high glucose group were mainly associated with IL-17 signaling pathway. Nonetheless, the DEGs of long-term high glucose group were primarily involved in IL-17 signaling pathway, MAPK signaling pathway, TNF signaling pathway, AGE-RAGE signaling pathway in diabetic complications, Toll-like receptor signaling pathway, and VEGF signaling pathway. Additionally, 8 hub genes of short-term high glucose group were enriched in metabolic pathway. Moreover, 10 hub genes of long-term high glucose group were enriched in ribosome pathway. Subsequently, in vitro experiment results found that high glucose can promote cell proliferation, and has a time accumulation effect. In addition, high glucose can induce the accumulation of inflammatory factors and promote angiogenesis. Collectively, these findings provide novel insights into the effect of diabetes mellitus type 2 (T2DM) on TNBC.

Inhibition of the neddylation E2 enzyme UBE2M in macrophages protects against E.coli-induced sepsis.

UBE2M, an essential neddylation E2 enzyme, has been implicated in the pathogenesis of various diseases, including cancers, viral infections, and obesity. However, whether UBE2M is involved in the pathogenesis of bacterial sepsis remains unclear. In an Escherichia coli (E.coli)-induced sepsis mouse model, increased UBE2M expression in macrophages in liver and lung tissues postinfection was observed. To further clarify the role of UBE2M in macrophages, mice with macrophage-specific deletion of UBE2M (Lysm+Ube2mf/f) were constructed. Compared with control mice, these mice presented decreased levels of proinflammatory cytokines, such as IL-1β, IL-6, and TNF-α; reduced sepsis-induced organ injury; and improved survival. Notably, macrophage-specific deletion of UBE2M did not impair E.coli clearance. Invitro experiments also revealed that UBE2M-deficient macrophages produced fewer proinflammatory cytokines after E.coli infection without hindering E.coli clearance. RNA-sequencing analysis revealed that UBE2M deletion in macrophages after lipopolysaccharide stimulation notably suppressed transcriptional activation within the JAK-STAT and Toll-like receptor signaling pathways, which was further confirmed by gene set enrichment analysis. Additionally, Western blotting results confirmed that UBE2M deletion inhibited the activation of the NF-κB, ERK, and JAK-STAT signaling pathways. In conclusion, our findings indicate that specific deletion of UBE2M in macrophages protects against E.coli-induced sepsis by downregulating the excessive inflammatory response, potentially providing a novel strategy against sepsis by targeting UBE2M.

Synthesis, characterization and biological profile of some new dihydropyrimidinone derivaties.

The rise of drug-resistant bacteria, viruses, and fungi has prompted the search for new drugs without cross-resistance to current treatments. As a result, the aim of this research was to synthesize various types of dihydropyrimidinones heterocyclic compounds and screened them for their antibiotic properties. Newly synthesized dihydropyrimidinone derivatives were characterized spectroscopically using proton NMR (1HNMR), and FT-IR. These substances were then subjected to molecular docking studies via Auto dock Vina software to determine their affinity for binding to proteins from different bacterial strains including (Staphylococcus epidermidis (S. epidermidis), Staphylococcus aureus (S. aureus), Mycobacterium luteus (M. luteus), Salmonella typhi (S.typhi), Bacillus subtilis (B. subtilis), and Escherichia coli (E. coli) and fungal (Candida glabrata (C. glabrata), Candida albicans (C. albicans), and Saccharomyces cerevisiae (S. cerevisiae) strains. Also in-vitro anti-fungal, anti-bacterial and anti-oxidant activity was performed by using ager well diffusion method and DPPH assay respectively. Moreover, the In-vivo biological evaluation of these derivatives was conducted by using carrageenan-induced hind paw model. The cytotoxicity profile of the synthesized derivatives was done via in-vitro MTT assay. All newly synthesized derivatives were confirmed via the multiple spectroscopic analysis techniques. All derivatives showed good binding affinities against the multiple targeted protein with. Compound 4c exhibited hightest potential with -10kcal/mol against bacterial strains. 4b showed best antifungal potential with -10.8kcal/mol binding affinities. For Bacillus subtilis compound 4b and 4c performed best with 17mm±2.21. for anti-fungal activity against Candida glabrata, amongst the five newly formed compounds, 4a showed best activity with 19mm±1.22 The analogue 4b exhibited best anti-oxidant potential with 63.85±1.39. Compound 4a and 4b showed highest anti-inflammatory potential with 1.011mm±0.247mg/kg and 1.447±0.212mg/kg in countering inflammation by targeting toll-like receptor activation and reduce the inflammation in hind paw edema. The selected derivatives exhibited no toxicity profile with 99 % and 98 % cell survival rate using compound 4a and 4b. Research has been done on the multiple biological activities of dihydropyrimidinones derivatives but the innovation on MDR is still pending. New dihydropyrimidinone derivatives were developed as agents to combat drug resistance. The results of these studies showed that newly synthesized dihydropyrimidinone derivatives are remarkably effective not only as anti-biotic agents but also counter inflammation caused by carrageenan resulting from the activation of the Toll-like receptors (TLRs) signaling pathway along with the non-toxic effect. So it is concluded that the recently synthesized new dihyropyrimidinone derivatives are highly effective antimicrobial derivatives with non-toxic effects on human cell lines.

Network pharmacology, transcriptomics, and biological validation reveal a lipid secretion inhibitory and anti-inflammatory mechanism of tanreqing gel in the treatment of acne.

Acne vulgaris is a common skin disease affecting the pilosebaceous unit, in which abnormal sebum secretion and inflammation play crucial roles. The traditional Chinese medicine Tanreqing has been utilized in dermatology to effectively treat various diseases. However, its effects and underlying mechanisms in acne vulgaris remain unclear. This study aims to assess the potential benefits of Tanreqing gel (TRQ) in acne treatment and to explore the mechanisms by which TRQ inhibits sebum secretion and reduces inflammation. A mouse model of acne induced by Cutibacterium acnes (C. acnes) was established. The impact of TRQ on acne lesions was assessed using optical imaging and histopathology. Network pharmacology and transcriptomics were used to identify significant intervention pathways and targets. Both in vivo and in vitro experiments were conducted to detect the expression of genes and proteins associated with inflammation and sebum metabolism. TRQ significantly improved pathological changes in the lesion areas of mice, such as redness, vascular dilation, and increased blood flow. It also reduced inflammatory cell infiltration in the dermis and inhibited the accumulation of lipids in the sebaceous glands. Network pharmacology analysis indicated that TRQ might exert anti-inflammatory effects through the IL-17, TOLL-like receptor, and NF-κB signaling pathways. The transcriptomic analysis confirmed the importance of these pathways in the C. acnes-induced acne model. Furthermore, TRQ was found to reduce sebum secretion by inhibiting fatty acid biosynthesis through the suppression of proteins in the PI3K-Akt signaling pathway. Cell experiments confirmed that TRQ could suppress the release of inflammatory factors induced by C. acnes surface structure peptidoglycan (PGN) and metabolite porphyrins. Additionally, it was observed to reverse the elevated porphyrin secretion associated with abnormal sebum production, ultimately relieving acne inflammation. This study demonstrated that TRQ effectively alleviates C. acnes-induced acne symptoms by inhibiting sebum secretion and inflammatory responses through multiple pathways and targets. It provides new insights and directions for acne treatment.

Transcriptomic and Microbiome Analyses of copepod Apocyclops royi in Response to an AHPND-causing strain of Vibrio parahaemolyticus

Copepods are small crustaceans that live in microorganism-rich aquatic environments and provide a key supply of live food for fish and shellfish larviculture. To better understand the host-pathogen interaction between the copepod and Vibrio parahaemolyticus causing acute hepatopancreatic necrosis disease (VPAHPND), the comparative transcriptome and microbiome analyses were conducted in copepod Apocyclops royi-TH following VPAHPND infection. Transcriptome analysis identified a total of 836 differentially expressed genes, with 275 upregulated and 561 downregulated genes. Subsequent analysis showed that a total of 37 differentially expressed genes were associated with the innate immune system, including 16 upregulated genes related to Toll-like receptor signaling pathway, antimicrobial peptides, and stress response genes, and 21 downregulated genes associated with immunological modulators, signaling molecules, and apoptosis-related proteins. Analysis of the copepod microbiome following VPAHPND infection showed that the microbes changed significantly after bacterial infection, with a reduced alpha diversity accompanied by the increased level of Proteobacteria and decreased levels of Bdellovibrionota, Bacteroidota, and Verrucomicrobiota. The population of Vibrio genera were increased significantly, while several other genera, including Denitromonas, Nitrosomonas, Blastopirellula, Fusibacter, Alteromonas, KI89A_clade, and Ruegeria, were decreased significantly after infection. These findings suggest that VPAHPND infection has a significant impact on the immune defense and the composition of the copepod microbiota.

Scientific knowledge about gene expression in ruminants under heat stress - A scientometric review.

Heat stress can alter the expression of genes in the individual's molecular response. The identification of these genes makes it possible to better understand the molecular response, identifying biomarker genes and indirect response pathways that can help with genetic improvement studies, animal welfare, separating more thermotolerant varieties and mitigating the effects of heat stress. The aim of this scientometric review was to characterize the state of the art of scientific research into gene expression in ruminants under heat stress, to define the most studied species, biology systems and genes, as well as the related biological pathways and processes. The articles for the dataset were compiled in the Web of Science database, refined individually and analyzed using the CiteSpace, RStudio, Excel and GraphPad Prism programs and the KEGG (Kyoto Encyclopedia of Genes and Genomes) database. The publications formed a data set containing 271 articles and an H-index of 37. The number of publications increased from 2011. The countries with the highest frequency of publications are India, the United States, China and Brazil, the ruminant species are cattle, buffaloes, sheep and goats, all zootechnical interest, and biology systems was reproduction, blood and lactation, due to the economic importance of the quality and quantity of production, to the ease of collecting and possibility of studies in vitro. Cattle have been extensively studied in comparison to other ruminants. The HSP70 gene has been the most studied, followed by the HSP family, HSF, BAX, TLR and BCL-2, these genes can be molecular markers of heat stress. The main pathways and biological processes of genes were in cattle the cancer pathway; in goats the Mixed, incl. myd88-dependent toll-like receptor signaling pathway, and lipopolys; in sheep the oxidoreductase; and in buffalo it was the BCL-2 family. The molecular responses are still recent and have not been established.

Peptostreptococcus Anaerobius enhances dextran sulfate sodium-induced colitis by promoting nf-κB-NLRP3-Dependent macrophage pyroptosis

ABSTRACT Evidence indicates that gut microbiota is crucial in ulcerative colitis (UC) development. Increased Peptostreptococcus species abundance is linked to UC, but its role and mechanisms in intestinal inflammation are not well understood. This study used a dextran sulfate sodium (DSS)-induced colitis model in mice, and different bacterial strains were administered via gavage. We assessed clinical manifestations, colonic barrier function, gut microbiota composition, and levels of inflammatory cytokines, NOD-like receptor family pyrin domain-containing 3 (NLRP3) signaling molecules, and pyroptosis-related proteins. Mouse bone marrow-derived macrophages (BMDMs) were infected with Peptostreptococcus anaerobius at different time points and multiplicities of infection (MOI). Cell viability and the expression of NLRP3 signaling molecules and pyroptosis-associated proteins were assessed. The inhibitors C29, TAK-242, and MCC950 were employed for Toll-like receptor (TLR) and NLRP3 signaling pathways. It was observed that P. anaerobius exacerbated intestinal inflammation and barrier injury in DSS-induced colitis in mice. Additionally, P. anaerobius contributed to gut microbiota dysbiosis during colitis progression. P. anaerobius induced the expression of NLRP3 signaling molecules and pyroptosis-associated proteins in mouse colitis tissues. In vitro assays demonstrated that P. anaerobius activated NLRP3 inflammasome and evoked gasdermin D-mediated pyroptosis and interleukin (IL)-1β secretion in macrophages. Furthermore, TLR2 and TLR4 were identified as key mediators of P. anaerobius-induced macrophage pyroptosis via activation of the Nuclear Factor-kappa B (NF-κB)-NLRP3 pathway. In conclusion, P. anaerobius promotes macrophage pyroptosis and IL-1β secretion through the TLR2/4-NF-κB-NLRP3 signaling axis, thereby aggravating colitis. P. anaerobius may represent a potential risk factor for UC development.

Transcriptome and Proteome Analyses Revealed Differences in JEV-Infected PK-15 Cells in Response to Ferroptosis Agonists and Antagonists.

Epidemic encephalitis B caused by Japanese encephalitis virus (JEV) is a common zoonotic disease that poses threats to both pigs and humans. The cellular defense mechanism is closely tied to the body's resistance to viral invasion. Regulated cell death, such as ferroptosis, is a strategy employed by host cells to defend against viral invasions. To understand the effect of ferroptosis on the proliferation of JEV, experimentally infected PK15 cells were treated with a ferroptosis agonist or antagonist. The results indicated that the ferroptosis agonist can suppress JEV proliferation, whereas the ferroptosis antagonist promotes JEV proliferation. Functional enrichment analysis showed that the ferroptosis agonist Erastin and antagonist SP600125 could affect JEV proliferation through the TNF, IL-17, Toll-like receptor, PI3K-AKT, and chemokine signaling pathways, as well as ECM-receptor interactions. Combined transcriptome and proteome analyses revealed 31 important genes, which are significantly associated with ferroptosis and the inflammatory response. Our results provide a better understanding of the molecular mechanisms through which ferroptosis affects the proliferation of JEV.

DNA and RNA Methylation in Periodontal and Peri-implant Diseases.

Periodontal and peri-implant diseases are primarily biofilm-induced pathologies in susceptible hosts affecting the periodontium and dental implants. Differences in disease susceptibility, severity, and patterns of progression have been attributed to immune regulatory mechanisms such as epigenetics. DNA methylation is an essential epigenetic mechanism governing gene expression that plays pivotal roles in genomic imprinting, chromosomal stability, apoptosis, and aging. Clinical studies have explored DNA methylation inhibitors for cancer treatment and predictive methylation profiles for disease progression. In periodontal health, DNA methylation has emerged as critical, evidenced by clinical studies unraveling its complex interplay with inflammatory genes and its regulatory role in periodontitis contributing to disease severity. Human studies have shown that methylation enzymes associated with gene reactivation (e.g., ten-eleven translocation-2) are elevated in periodontitis compared with gingivitis. Dysregulation of these genes can lead to the production of inflammatory cytokines and an altered initial response to bacteria via the toll-like receptor signaling pathway in periodontal diseases. In addition, in peri-implant diseases, this dysregulation can result in altered DNA methylation levels and enzymatic activity influenced by the properties of the titanium surface. Beyond traditional perspectives, recent evidence highlights the involvement of RNA methylation (e.g., N6-methyladenosine [m6A], N6,2'-0-dimethyladenosine [m6Am]) in periodontitis and peri-implantitis lesions, playing vital roles in the innate immune response, production of inflammatory cytokines, and activation of dendritic cells. Both DNA and RNA methylation can influence the gene expression, virulence, and bacterial behavior of well-known periodontal pathogens such as Porphyromonas gingivalis. Alterations in bacterial methylation patterns result in changes in the metabolism, drug resistance, and gene expression related to survival in the host, thereby promoting tissue degradation and chronic inflammatory responses. In summary, the present state-of-the-art review navigates the evolving landscape of DNA and RNA methylation in periodontal and peri-implant diseases, integrating recent developments and mechanisms to reshape the understanding of epigenetic dynamics in oral health.

Deciphering the pharmacological mechanism of Radix astragali for allergic rhinitis through network pharmacology and experimental validation

Radix Astragali (RA) has been recognized for its therapeutic potential in allergic rhinitis (AR), yet its potential pharmacological mechanisms remain elusive. This study systematically investigated the physicochemical properties and biological activities of RA’s phytochemicals, aiming to elucidate their targets and mechanisms in AR treatment. We identified 775 potential targets of RA’s key phytochemicals and intersected these with 29,544 AR-related disease targets, pinpointing 747 shared therapeutic targets. A protein-protein interaction network analysis categorized these targets into five subclusters, with TNF, NFKB1, IKBKB, NFKBIA, and CHUK emerging as central nodes. Enrichment analysis revealed their roles in inflammatory and immune responses, particularly through the NF-κB, TNF, IL-17, Toll-like receptor, and NOD-like receptor signaling pathways. Molecular docking and dynamics simulations confirmed the strong binding affinity and stability of RA’s phytochemicals to these targets. In vivo, RA intervention effectively reversed the expression of key inflammatory markers in an IL-13-induced nasal mucosa inflammation model. Our findings suggest that RA’s multitargeted approach involves the modulation of critical inflammatory pathways, highlighting its therapeutic potential.

Comprehensive analysis of differentially expressed genes in Toll-Like Receptor Signaling Pathway: Insights into New-Onset Microscopic Polyangiitis.

We aimed to elucidate the potential contributions of the toll-like receptor (TLR) signalling pathway and identify promising candidates for new-onset microscopic polyangiitis (MPA) using integrated bioinformatics analysis. A PCR array was used to determine the expression profiles of TLR signalling-related genes in CD4+T lymphocytes of individuals with new-onset MPA and healthy controls. Four genes were selected for validation through real-time quantitative polymerase chain reaction (RT-qPCR). Followed by functional enrichment and pathway analysis, we identified the hub genes with cytoHubba. The differentially expressed miRNAs of the target genes were subsequently predicted and visualized via Cytoscape. Finally, these candidates were validated and evaluated at the expression level and for diagnostic value in public databases. Nineteen differentially expressed genes were screened, and the levels of the validated genes detected using RT‒qPCR were consistent with the findings obtained through the PCR array. The significantly enriched signalling pathways involved were TLR signalling pathway, IL-17 signalling pathway, and NF-κB signalling pathway. Nine hub genes and nine key miRNAs were identified. Furthermore, analysis of three distinct gene expression datasets validated several key genes (TLR4, MYD88, IRF1, CXCL10, CXCL8, and CSF2), showing significant differences between groups and strong diagnostic value, especially TLR4, MYD88, and IRF1. Interestingly, in contrast to the validation results, our results showed that CXCL10 and CXCL8 expression levels were markedly lower, but CSF2 was highly expressed in patients with MPA compared to controls. Aberrant expression of TLRs may occur in CD4+ T lymphocytes of patients with new-onset MPA, offering insights into the pathogenesis as well as potential biomarkers and novel therapeutic targets.

Mechanistic Evaluation of EGCG against Atopic Dermatitis: An in Vivo and in Silico Study

Background: Atopic dermatitis, often referred to as AD, is a common chronic and relapsing skin disease with few treatment options. Epigallocatechin 3-gallate (EGCG), the most abundant catechin derived from green tea, has gained widespread attention due to its excellent anti-inflammatory and anti-oxidative properties. This research aims to investigate the effect and mechanism of EGCG in alleviating AD by integrative approach of an in vivo AD mouse model, network pharmacology and molecular docking. Materials and Methods: EGCG was administered by intragastric administration at the dose of 100 mg/kg, once daily for 14 consecutive days to AD mice, a model established by repeated application of 2, 4-dinitrofluorobenzene (DNFB) to the mouse dorsal skin. Results: Macroscopical and histopathological examination showed that oral administration with EGCG could markedly relieve AD-like skin phenotypes, as evidenced by a substantial reduction in severity score, serum immunoglobulin E (IgE) level, epidermal thickness, and the infiltration of inflammatory cells and mast cells into the dermal layers. Mechanistically, the network pharmacology analysis indicated that the protective effects of EGCG against AD were mainly related to the regulation of AGE-RAGE signaling pathway in diabetic complications, Th17 cell differentiation, IL-17 signaling pathway, NF-κB signaling pathway, Toll-like receptor signaling pathway, TNF signaling pathway, etc. In silico molecular docking demonstrated the high binding affinity of EGCG to the core targets such as ABCB1, MMP2, MMP9, MAPK14 and STAT1. Furthermore, western blotting (WB) results showed that EGCG obviously downregulated the expression of RAGE and STAT1, thereby regulating the AGE-RAGE signaling pathway to improve AD. Conclusions: These findings indicated that EGCG can be a promising therapeutic candidate for AD.

Network Pharmacology and Bioinformatics Analyses Identify the Core Genes and Pyroptosis-Related Mechanisms of Nardostachys Chinensis for Atrial Fibrillation.

Nardostachys chinensis is an herbal medicine widely used in the treatment of atrial fibrillation (AF), but the mechanism is unclear. To explore the molecular mechanism of N. chinensis against AF. The TCMSP was used to screen the active N. chinensis compounds and their targets. Differentially expressed genes (DEGs) for AF were identified using open-access databases. Using Venn diagrams, the cross-targets of N. chinensis, pyroptosis, and AF were obtained. The genes underwent molecular docking as well as gene set enrichment analysis (GSEA). A nomogram based on candidate genes was constructed and evaluated with the clinical impact curve. After that, the immune infiltration of the dataset was analyzed by single sample GSEA (ssGSEA). Finally, microRNAs (miRNAs) and transcription factors (TFs) were predicted based on candidate genes. Tumor necrosis factor (TNF) and caspase-8 (CASP8) were obtained as candidate genes by taking the intersection of DEGs, targets of N. chinensis, and pyroptosis-related genes. Tolllike receptor (TLR) and peroxisome proliferator-activated receptor (PPAR) signaling pathways were linked to candidate genes. Additionally, immune cell infiltration analysis revealed that CASP8 was associated with natural killer T cells, natural killer cells, regulatory T cells (Tregs), myeloid-derived suppressor cells (MDSC), macrophages, CD8 T cells, and CD4 T cells. Finally, miR-34a-5p and several TFs were found to regulate the expression of CASP8 and TNF. CASP8 and TNF are potential targets of N. chinensis intervention in pyroptosisrelated AF, and the TLR/NLRP3 signaling pathway may be associated with this process.

Therapeutic Correlation of TLR-4 Mediated NF-κB Inflammatory Pathways in Ischemic Injuries.

Ischemia-reperfusion (I/R) injury refers to the tissue damage that happens when blood flow returns to tissue after a period of ischemia. I/R injuries are implicated in a large array of pathological conditions, such as cerebral, myocardial, renal, intestinal, retinal and hepatic ischemia. The hallmark of these pathologies is excessive inflammation. Toll-like receptors (TLRs) are recognized as significant contributors to inflammation caused by pathogens and, more recently, inflammation caused by injury. TLR-4 activation initiates a series of events that results in activation of nuclear factor kappa-B (NF-κB), which stimulates the production of pro-inflammatory cytokines and chemokines, exacerbating tissue injury. Therefore, through a comprehensive review of current research and experimentation, this investigation elucidates the TLRs signalling pathway and the role of TLR-4/NF-κB in the pathophysiology of I/R injuries. Furthermore, this review highlights the various pharmacological agents (TLR-4/NF-κB inhibitors) with special emphasis on the various ischemic injuries (cerebral, myocardial, renal, intestinal, retinal and hepatic). Future research should prioritise investigating the specific molecular pathways that cause TLR-4/NF-κBmediated inflammation in ischemic injuries. Additionally, efforts should be made to enhance treatment approaches in order to enhance patient outcomes.