Lipopolysaccharide Exposure Research Articles

Transcript and Lipid Profile Alterations in Astrocyte-Neuron Mitochondrial Transfer Under Lipopolysaccharide Exposure: An InVitro Study.

Sepsis-associated encephalopathy (SAE) is a brain dysfunction for which no effective therapy currently exists. Recent studies suggest that transferring mitochondria from astrocytes to neurons may benefit SAE patients, though the underlying mechanism remains unclear. We cultured astrocytes and neurons from mice invitro. Astrocytes were stimulated with lipopolysaccharide (LPS) for 24 h, and the astrocyte-conditioned medium (ACM) was collected. Neuronal cultures were then treated with ACM or mitochondria-depleted ACM (mdACM) for further analysis. Mitochondrial transfer was examined under a fluorescence microscope. Western blotting analyzed the protein expression of genes related to apoptosis and mitochondrial metabolism. RNA sequencing and mass spectrometry were employed to investigate the mechanisms underlying mitochondrial transfer. Astrocyte-derived mitochondria migrated toward and connected with LPS-exposed neurons. The addition of ACM significantly attenuated LPS-induced alterations in the proteins linked to apoptosis and mitochondrial dynamics. RNA sequencing revealed notable alterations in the transcript profile of neurons upon ACM treatment, highlighting the involvement of mitochondria metabolism, inflammation, and apoptosis-related factors. Additionally, mitochondrial transfer modified the lipid composition of neurons, increasing phosphatidylserine levels, which correlated with neuroinflammation and enriched pathways related to cytokine and MAPK signaling. Our findings suggest that astrocyte-neuron mitochondrial transfer holds therapeutic potential for alleviating SAE, possibly through the anti-inflammatory effects of lipids, particularly phosphatidylserine.

Imaging flow cytometry reveals LPS-induced changes to intracellular intensity and distribution of α-synuclein in a TLR4-dependent manner in STC-1 cells

BackgroundParkinson’s disease is a chronic neurodegenerative disorder, where pathological protein aggregates largely composed of phosphorylated α-synuclein are implicated in disease pathogenesis and progression. Emerging evidence suggests that the interaction between pro-inflammatory microbial factors and the gut epithelium contributes to α-synuclein aggregation in the enteric nervous system. However, the cellular sources and mechanisms for α-synuclein pathology in the gut are still unclear. MethodsThe STC-1 cell line, which models an enteroendocrine population capable of communicating with the microbiota, immune and nervous systems, was treated with a TLR4 inhibitor (TAK-242) prior to microbial lipopolysaccharide (LPS) exposure to investigate the role of TLR4 signalling in α-synuclein alterations. Antibodies targeting the full-length protein (α-synuclein) and the Serine-129 phosphorylated form (pS129) were used. Complex, multi-parametric image analysis was conducted through confocal microscopy (with Zen 3.8 analysis) and imaging flow cytometry (with IDEAS® analysis). ResultsConfocal microscopy revealed heterogenous distribution of α-synuclein and pS129 in STC-1 cells, with prominent pS129 staining along cytoplasmic processes. Imaging flow cytometry further quantified the relationship between various α-synuclein morphometric features. Thereafter, imaging flow cytometry demonstrated a dose-specific effect of LPS, where the low (8 μg/mL), but not high dose (32 μg/mL), significantly altered measures related to α-synuclein intensity, distribution, and localisation. Pre-treatment with a TLR4 inhibitor TAK-242 alleviated some of these significant alterations. ConclusionThis study demonstrates that LPS-TLR4 signalling alters the intracellular localisation of α-synuclein in enteroendocrine cells in vitro and showcases the utility of combining imaging flow cytometry to investigate subtle protein changes that may not be apparent through confocal microscopy alone. Further investigation is required to understand the apparent dose-dependent effects of LPS on α-synuclein in the gut epithelium in healthy states as well as conditions such as Parkinson’s disease.

D-sodium lactate promotes the activation of NF-κB signaling pathway induced by lipopolysaccharide via histone lactylation in bovine mammary epithelial cells.

Lactate is a glycolytic end product that is further metabolized as an energy source. This end product has been associated with certain diseases, including sepsis and tumors, and it can regulate the transition of macrophages to an anti-inflammatory state. This study aimed to explore the effects of lactate on the inflammatory responses of mammary gland epithelial cells, which constitute the first line of defense against pathogens in mammary glands. Bovine mammary epithelial cells (BMECs) were challenged with lipopolysaccharide (LPS) in the presence or absence of D-sodium lactate (D-nala). LPS exposure increased the concentration of lactate both inside and outside the cells. Further, inhibiting glycolysis diminished the LPS-induced production of proinflammatory cytokines. Treatment with LPS, exogenous D-nala, and their combination upregulated the expression levels of MCT1, increased the intracellular levels of lactate and histone H3 lysine 18 lactylation (H3K18la), and activated the nuclear factor kappa-light-chain-enhancer of activated B cell (NF-κB) signaling pathway. The lactylation of H3K18 was mediated by p300/CBP. The p300/CBP inhibitor C646 decreased the level of H3K18la, reversing the activation of the NF-κB signaling pathway and release of proinflammatory cytokines. Therefore, LPS increased the intracellular level of lactate by upregulating MCT1 and glycolysis. D-nala exacerbated the LPS-induced inflammatory responses in BMECs. Moreover, intracellular lactate enhanced the activation of the NF-κB signaling pathway through the p300/CBP-mediated lactylation of H3K18. Thus, the findings of this study expand our understanding of lactate function in immune regulation.

Genetic variation in IL-4 activated tissue resident macrophages determines strain-specific synergistic responses to LPS epigenetically

How macrophages in the tissue environment integrate multiple stimuli depends on the genetic background of the host, but this is still poorly understood. We investigate IL-4 activation of male C57BL/6 and BALB/c strain specific in vivo tissue-resident macrophages (TRMs) from the peritoneal cavity. C57BL/6 TRMs are more transcriptionally responsive to IL-4 stimulation, with induced genes associated with more super enhancers, induced enhancers, and topologically associating domains (TAD) boundaries. IL-4-directed epigenomic remodeling reveals C57BL/6 specific enrichment of NF-κB, IRF, and STAT motifs. Additionally, IL-4-activated C57BL/6 TRMs demonstrate an augmented synergistic response upon in vitro lipopolysaccharide (LPS) exposure, despite naïve BALB/c TRMs displaying a more robust transcriptional response to LPS. Single-cell RNA sequencing (scRNA-seq) analysis of mixed bone marrow chimeras indicates that transcriptional differences and synergy are cell intrinsic within the same tissue environment. Hence, genetic variation alters IL-4-induced cell intrinsic epigenetic reprogramming resulting in strain specific synergistic responses to LPS exposure.

Understanding the neurobiological mechanisms of LPS‑induced memory impairment.

In recent years, growing evidence suggests that lipopolysaccharide (LPS), a bacterial endotoxin found in the outer membrane of gram‑negative bacteria, can influence cognitive functions, particularly memory formation and retrieval. However, the underlying mechanisms through which LPS exerts its effects on memory remain incompletely understood. This review used various electronic databases, including PubMed, Scopus, and Web of Science, to identify relevant studies published between 2000 and 2024. Articles were selected based on their focus on LPS‑induced memory impairments, including experimental models, molecular pathways, and neurochemical alterations. LPS administration has been consistently shown to disrupt memory processes in both animals and humans, although the magnitude and duration of memory impairments might vary depending on factors such as dose, timing, and context of LPS exposure. Several potential mechanisms have been proposed to explain LPS‑induced memory deficits, including neuroinflammation, alterations in synaptic plasticity, disruption of neurotransmitter systems, and dysfunction of the blood‑brain barrier. Moreover, LPS has been found to activate immune signaling pathways, such as toll‑like receptors, interleukins, and microglia, which can further contribute to cognitive impairments. Such insights may pave the way for the development of targeted therapeutic interventions aimed at ameliorating memory deficits associated with conditions involving LPS exposure, including bacterial infections, sepsis, and neuroinflammatory disorders.

Astrocyte-Derived Interleukin 11 Modulates Astrocyte-Microglia Crosstalk via Nuclear Factor-κB Signaling Pathway in Sepsis-Associated Encephalopathy.

Sepsis-associated encephalopathy (SAE) is a severe and frequent septic complication, characterized by neuronal damage as key pathological features. The astrocyte-microglia crosstalk in the central nervous system (CNS) plays important roles in various neurological diseases. However, how astrocytes interact with microglia to regulate neuronal injury in SAE is poorly defined. In this study, we aim to investigate the molecular basis of the astrocyte-microglia crosstalk underlying SAE pathogenesis and also to explore the new therapeutic strategies targeting this crosstalk in this devastating disease. We established a human astrocyte/microglia coculture system on a microfluidic device, which allows real-time and high-resolution recording of glial responses to inflammatory stimuli. Based on this microfluidic system, we can test the responses of astrocytes and microglia to lipopolysaccharide (LPS) treatment, and identify the molecular cues that mediate the astrocyte-microglia crosstalk underlying the pathological condition. In addition, the SAE mouse model was utilized to determine the state of glial cells and evaluate the therapeutic effect of drugs targeting the astrocyte-microglia crosstalk invivo. Here, we found that activated astrocytes and microglia exhibited close spatial interaction in the SAE mouse model. Upon LPS exposure for astrocytes, we detected that more microglia migrated to the central astrocyte culture compartment on the microfluidic device, accompanied by M1 polarization and increased cell motility in microglia. Cytokine array analysis revealed that less interleukin 11 (IL11) was secreted by astrocytes following LPS treatment, which further promoted reprogramming of microglia to pro-inflammatory M1 phenotype via the nuclear factor-κB (NF-κB) signaling pathway. Intriguingly, we found that IL11 addition markedly rescued LPS-induced neuronal injuries on the microfluidic system and brain injury in the SAE mouse model. This study defines an unknown crosstalk of astrocyte-microglia mediated by IL11, which contributed to the neuropathogenesis of SAE, and suggested a potential therapeutic value of IL11 in the devastating disease.

Effects of maternal Escherichia coli lipopolysaccharide exposure on offspring: insights from lncRNA analysis in laying hens

Parental living environment significantly impacts on offspring, yet related studies are lacking in livestock and poultry production. The present study found that lipopolysaccharide (LPS, Escherichia coli, 0.2 mg/kg) stimulation in F0 hens led to growth retardation and a decrease in egg-laying rate in the unchallenged F1 hens. Using strand-specific transcriptomic data of peripheral blood mononuclear cells (PBMCs) in F1 hens, we identified 100 differentially expressed lncRNAs (DELs) and 452 differentially expressed genes (DEGs). LPS primarily affected the metabolic pathways of the offspring, possibly reducing the egg-laying rate of the F1 hens by inhibiting the ferroptosis signaling pathway and the expression of DEGs involved, such as NCOA4, SLC40A1, STEAP3, and TFRC. Using Pearson correlation analysis, we constructed a lncRNA-mRNA-egg-laying rate regulation network and found that the newly identified lncRNA MSTRG.6500.1 and its positively regulated target genes (ENSGALT00000051184, ENSGALT00000053276, NPPA, OSBP2, and TRARG1) were significantly downregulated in the F1 LPS group, which might be the main reason for the decrease in egg-laying rate of the LPS group. These results provide important references for the study of growth and reproductive performance in laying hens, revealing the impact of parental living environment on animal health and production performance, and providing a theoretical basis for future related research and breeding practices.

EP300-mediated H3 acetylation elevates MTHFD2 expression to reduce mitochondrial dysfunction in lipopolysaccharide-induced tubular epithelial cells

Sepsis represents an organ dysfunction resulting from the host’s maladjusted response to infection, and can give rise to acute kidney injury (AKI), which significantly increase the morbidity and mortality of septic patients. This study strived for identifying a novel therapeutic strategy for patients with sepsis-induced AKI (SI-AKI). Rat tubular epithelial NRK-52E cells were subjected to lipopolysaccharide (LPS) exposure for induction of in-vitro SI-AKI. The expressions of E1A binding protein p300 (EP300) and methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) in NRK-52E cells were assessed by western blot and qRT-PCR, and their interaction was explored by chromatin immunoprecipitation performed with antibody for H3K27 acetylation (H3K27ac). The effect of them on SI-AKI-associated mitochondrial dysfunction of tubular epithelial cells was investigated using transfection, MTT assay, TUNEL staining, 2′,7′-Dichlorodihydrofluorescein diacetate probe assay, Mitosox assay, and JC-1 staining. MTHFD2 and EP300 were upregulated by LPS exposure in NRK-52E cells. LPS increased the acetylation of H3 histone in the MTHFD2 promoter region, and EP300 suppressed the effect of LPS. EP300 ablation inhibited the expression of MTHFD2. MTHFD2 overexpression antagonized LPS-induced viability reduction, apoptosis promotion, reactive oxygen species overproduction, and mitochondrial membrane potential collapse of NRK-52E cells. By contrast, MTHFD2 knockdown and EP300 ablation brought about opposite consequences. Furthermore, MTHFD2 overexpress and EP300 ablation counteracted each other’s effect in LPS-exposed NRK-52E cells. EP300-mediated H3 acetylation elevates MTHFD2 expression to reduce mitochondrial dysfunction of tubular epithelial cells in SI-AKI.

Mogroside V protects lipopolysaccharides-induced lung inflammation chicken via suppressing inflammation mediated by the Th17 through the gut-lung axis.

Lipopolysaccharide (LPS) exposure triggers pulmonary inflammation, leading to compromised lung function in broiler. As amplified by policy restrictions on antibiotic usage, seeking antibiotic alternatives has become imperative. Mogroside V (MGV) has been reported to have a beneficial role in livestock and poultry production due to its remarkable antiinflammatory effects. Despite evidence showcasing MGV's efficacy against LPS-triggered lung inflammation, its precise mechanism of action remains elusive. In this study, we transplanted normal fecal microbiota (CF), fecal microbiota modified by MGV (MF), and sterile fecal filtrate (MS) into broiler with LPS-induced pneumonia. The results showed that through fecal microbiota transplantation (FMT), transplanting MGV-induced microbial populations significantly mitigated tissue damage induced by LPS and enhanced the mRNA level of pulmonary tight junction proteins and mucoprotein (P < 0.01). The expression levels of RORα (P < 0.001), Foxp3 (P < 0.01), and PD-L1 (P < 0.01) were significantly increased in the MF group than CF group. The concentrations of IL-6 and IL-17 in broilers lung tissue of MF group were lower than those in broilers of CF group (P < 0.05). Furthermore, the concentration of TGF-β in broilers serum of MS and MF groups was higher than those in broilers of CF group (P < 0.05). Microbial community analysis demonstrated that at genus level, the harmful bacterial populations Escherichia-Shigella and Helicobacter following FMT treatment were significantly reduced in MF group (P < 0.05), potentially mediating its protective effects. Compared with CF group, valerate content and FFAR2 mRNA expression levels in MF group were significantly increased (P < 0.05). The study suggests that MGV via the gut-lung axis, attenuates Th17-mediated inflammation, offering promise as a therapeutic strategy against LPS-induced lung inflammation in chickens.

Unveiling the role of lipopolysaccharide-related genes in diabetic retinopathy: identification of key biomarkers and immune infiltration analysis

BackgroundGrowing evidence suggests a link between systemic lipopolysaccharide (LPS) exposure and worsening diabetic retinopathy (DR). This study aims to investigate DR’s pathogenesis by analyzing LPS-related genes (LRGs) through bioinformatics.MethodsThe CTD database was utilized to identify LRGs. The datasets associated with DR were acquired from the GEO database. The Venn diagram was used to identify the differentially expressed LRGs (DLRGs), and the putative molecular mechanism of these DLRGs was investigated through functional enrichment analysis. We used WGCNA, Lasso regression, and RF to identify hub DLRGs. The expression levels of these hub DLRGs were validated in an independent dataset (GSE102485) and cell experiments. Employing the CIBERSORT algorithm, we examined the infiltration of 22 distinct immune cell types in DR and assessed the association between key DLRGs and immune infiltrates through correlation analysis.ResultsA total of 71 DLRGs were detected. These genes exhibited significant enrichment in pathways associated with inflammation. In addition, the in-depth analysis uncovered that five hub DLRGs (STK33 and EPHX2) linked to bacterial LPS displayed noteworthy diagnostic potential for individuals diagnosed with DR. The hub DLRGs expression in the high glucose-induced DR model was confirmed by qRT-PCR analysis. Furthermore, examination of immune infiltration indicated a significant association between these five genes and the extent of immune cell infiltration.ConclusionSTK33 and EPHX2 serve as biomarkers related to bacterial LPS. Exploring these genes in-depth could provide innovative ideas and a foundation for comprehending the progression of the disease and developing targeted treatments for DR.

SIRT5-mediated HOXA5 desuccinylation inhibits ferroptosis to alleviate sepsis induced-lung injury.

Acute lung injury (ALI) is a common and severe complication of sepsis with a high mortality rate. Ferroptosis, an iron-dependent form of cell death, contributes to lung injury. Homeobox A5 (HOXA5) is involved in the regulation of septic acute kidney damage; however, its function on ferroptosis in septic ALI remains unclear. An in vitro model of septic lung injury was established in the pulmonary epithelial cell line (MLE-12) via lipopolysaccharide (LPS) stimulation. Cell viability, ferrous iron (Fe2+) level, and cellular lipid reactive oxygen species (ROS) were determined with Cell Counting Kit-8 assay, iron assay kit, and BODIPY™ 665/676 molecular probe, respectively. HOXA5, ferroptosis suppressor protein 1 (FSP1), sirtuin 5 (SIRT5), and glutathione peroxidase 4 (GPX4) expressions were measured using western blotting and Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR. Chromatin immunoprecipitation and luciferase reporter assays were performed to validate HOXA5 binding to the FSP1/GPX4 promoter, and regulation of SIRT5 on HOXA5 desuccinylation was confirmed through co-immunoprecipitation. LPS stimulation induced ferroptosis (reduced cell viability, elevated Fe2+ and lipid ROS levels, and decreased GPX4 levels) and downregulated FSP1 and HOXA5 protein levels. HOXA5 overexpression neutralized LPS-induced ferroptosis. Moreover, LPS exposure inhibited HOXA5 binding to the FSP1 promoter, which was counteracted via HOXA5 overexpression. Furthermore, SIRT5 overexpression suppressed LPS-induced ferroptosis. In LPS-challenged MLE-12 cells, SIRT5-mediated HOXA5 desuccinylation was reduced. HOXA5 depletion neutralized the suppressive role of SIRT5 overexpression in LPS-induced ferroptosis. SIRT5-mediated HOXA5 desuccinylation inhibited LPS-induced ferroptosis by upregulating FSP1, which may offer a prospective therapeutic strategy for septic lung injury.

Identification of Potential Sepsis Therapeutic Drugs Using a Zebrafish Rapid Screening Approach.

In the military, combat wound infections can progress rapidly to life-threatening sepsis. The discovery of effective small-molecule drugs to prevent and/or treat sepsis is a priority. To identify potential sepsis drug candidates, we used an optimized larval zebrafish model of endotoxicity/sepsis to screen commercial libraries of drugs approved by the U.S. Food and Drug Administration (FDA) and other active pharmaceutical ingredients (APIs) known to affect pathways implicated in the initiation and progression of sepsis in humans (i.e., inflammation, mitochondrial dysfunction, coagulation, and apoptosis). We induced endotoxicity in 3- and 5-day post fertilization larval zebrafish (characterized by mortality and tail fin edema (vascular leakage)) by immersion exposure to 60 µg/mL Pseudomonas aeruginosa lipopolysaccharide (LPS) for 24 h, then screened for the rescue potential of 644 selected drugs at 10 µM through simultaneous exposure to LPS. After LPS exposure, we used a neurobehavioral assay (light-dark test) to further evaluate rescue from endotoxicity and to determine possible off-target drug side effects. We identified 29 drugs with > 60% rescue of tail edema and mortality. Three drugs (Ketanserin, Tegaserod, and Brexpiprazole) produced 100% rescue and did not differ from the controls in the light-dark test, suggesting a lack of off-target neurobehavioral effects. Further testing of these three drugs at a nearly 100% lethal concentration of Klebsiella pneumoniae LPS (45 µg/mL) showed 100% rescue from mortality and 88-100% mitigation against tail edema. The success of the three identified drugs in a zebrafish endotoxicity/sepsis model warrants further evaluation in mammalian sepsis models.

Interaction of Liposomes Containing the Carrageenan/Echinochrome Complex with Human HaCaT Keratinocytes In Vitro.

Liposomal drug delivery systems are successfully used in various fields of medicine for external and systemic applications. Marine organisms contain biologically active substances that have a unique structure and exhibit a wide range of biological activities. Polysaccharide of red seaweed (carrageenan (CRG)), and water-insoluble sea urchin pigment (echinochrome (Ech)) interact with each other and form a stable complex. We included the CRG/Ech complex in liposomes for better permeability into cells. In our research, tetramethylrhodamine isothiocyanate TRITC-labeled CRG was synthesized to study the interaction of the complex encapsulated in liposomes with human epidermal keratinocytes (HaCaTs) widely used to expose the skin to a variety of substances. Using confocal microscopy, we found that liposomes were able to penetrate HaCaT cells with maximum efficiency within 24 h, and pre-incubation of keratinocytes with liposomes resulted in the delivery of the CRG/Ech complex into the cytoplasm. We investigated the anti-inflammatory effects of liposomes, including the lysosomal regulation, increased intracellular ROS levels, and increased NO synthesis in lipopolysaccharide (LPS)- or Escherichia coli (E. coli)-induced inflamed skin cells. Liposomes containing the CRG/Ech complex significantly reduced lysosomal activity by 26% in LPS-treated keratinocytes and decreased ROS levels in cells by 23% after LPS exposure. It was found that liposomes with the complex improved the migration of HaCaT keratinocytes incubated with high-dose LPS by 47%. The results of the work, taking into account the good permeability of liposomes into keratinocytes, as well as the anti-inflammatory effect on cells treated with LPS or E. coli, show the prospects of using liposomes containing the CRG/Ech complex as an anti-inflammatory agent in the fight against skin infections.

Impact of Oxidative Stress and Neuroinflammation on Sarco/Endoplasmic Reticulum Ca2+-ATPase 2b Downregulation and Endoplasmic Reticulum Stress in Temporal Lobe Epilepsy.

Epilepsy is one of the most common neurological disorders. Calcium dysregulation and neuroinflammation are essential and common mechanisms in epileptogenesis. Sarco/endoplasmic reticulum (ER) Ca2+-ATPase 2b (SERCA2b), a crucial calcium regulatory pump, plays pathological roles in various calcium dysregulation-related diseases. However, the link between SERCA2b and neuroinflammation in epilepsy remains undetermined. This study aimed to establish the relationship between SERCA2b, oxidative stress, and neuroinflammation in epilepsy to elucidate the underlying molecular mechanism in epileptogenesis. Neuroinflammation and oxidative stress were induced in N2a cells using lipopolysaccharide (LPS) and hydrogen peroxide (H2O2). However, experimental temporal lobe epilepsy (TLE) was induced in mice using pilocarpine. Further, effects of oxidative stress and neuroinflammation on SERCA2b and ER stress markers were assessed at protein and mRNA levels. Calcium imaging was employed to determine intracellular calcium levels. SERCA2b expression significantly decreased after LPS, H2O2, and pilocarpine exposure at both mRNA and protein levels, mediated by upregulating neuroinflammation. This downregulation of SERCA2b was associated with increased production of reactive oxygen species and elevated intracellular calcium levels, leading to elevated ER stress markers. Our findings highlight a link between oxidative stress, neuroinflammation and SERCA2b in TLE. The results suggest that targeting SERCA2b could restore calcium homeostasis and ER stress processes, potentially providing a therapeutic option for TLE. This study underscores the importance of SERCA2b in the pathophysiology of epilepsy and its potential as a therapeutic target.

Bacterial products initiation of alpha-synuclein pathology: an in vitro study

Parkinson’s Disease (PD) is a prevalent and escalating neurodegenerative disorder with significant societal implications. Despite being considered a proteinopathy, in which the aggregation of α-synuclein is the main pathological change, the intricacies of PD initiation remain elusive. Recent evidence suggests a potential link between gut microbiota and PD initiation, emphasizing the need to explore the effects of microbiota-derived molecules on neuronal cells. In this study, we exposed dopaminergic-differentiated SH-SY5Y cells to microbial molecules such as lipopolysaccharide (LPS), rhamnolipid, curli CsgA and phenol soluble modulin α-1 (PSMα1). We assessed cellular viability, cytotoxicity, growth curves and α-synuclein levels by performing MTS, LDH, real-time impedance readings, qRT-PCR and Western Blot assays respectively. Statistical analysis revealed that rhamnolipid exhibited concentration-dependent effects, reducing viability and inducing cytotoxicity at higher concentrations, increasing α-synuclein mRNA and protein levels with negative effects on cell morphology and adhesion. Furthermore, LPS exposure also increased α-synuclein levels. Curli CsgA and PSMα-1 showed minimal or no changes. Our findings suggest that microbiota-derived molecules, particularly rhamnolipid and LPS, impact dopaminergic neurons by increasing α-synuclein levels. This study highlights the potential involvement of gut microbiota in initiating the upregulation of α-synuclein that may further initiate PD, indicating the complex interplay between microbiota and neuronal cells.

Mathematical model of the inflammatory response to acute and prolonged lipopolysaccharide exposure in humans

One in five deaths worldwide is associated with sepsis, which is defined as organ dysfunction caused by a dysregulated host response to infection. An increased understanding of the pathophysiology of sepsis could provide improved approaches for early detection and treatment. Here we describe the development and validation of a mechanistic mathematical model of the inflammatory response, making use of a combination of in vitro and human in vivo data obtained from experiments where bacterial lipopolysaccharide (LPS) was used to induce an inflammatory response. The new model can simulate the responses to both acute and prolonged inflammatory stimuli in an experimental setting, as well as the response to infection in the clinical setting. This model serves as a foundation for a sepsis simulation model with a potentially wide range of applications in different disciplines involved with sepsis research.

Hepatic Gene Expression Changes of Zebrafish Fed Yeast Prebiotic, Yeast Probiotic, Black Soldier Fly Meal, and Butyrate

As global fish consumption rises, improving fish health through immunomodulatory feed ingredients shows promise while also supporting growth performance. This study investigated the effects of yeast prebiotics, probiotics, a postbiotic (butyrate), and black soldier fly larvae (BSFL) meal on fish immune responses. Zebrafish were fed diets containing these ingredients for 63 days and then exposed to either Pseudomonas aeruginosa lipopolysaccharide (LPS) or live Flavobacterium psychrophilum to assess hepatic candidate gene expression and weight gain. No mortalities were observed post-immune challenges, and weight gains were not significantly different across treatments. Liver samples were collected for mRNA analysis, and real-time qPCR was used to evaluate the expression of immune-related genes such as TNF-α, IL-1β, hepcidin, and NF-κB/p65. NF-κB/p65 was upregulated in response to immune challenges, indicating a reaction to both LPS and pathogen exposure. Fish on the BSFL diet showed decreased NF-κB/p65 expression after the pathogen challenge, while probiotic-fed fish had reduced angiopoietin-like 4 (angptl4) levels following LPS exposure. Butyrate supplementation had the most significant impact, downregulating pro-inflammatory cytokines and other immune-related genes, suggesting a protective effect. These findings support the health benefits of BSFL and sodium butyrate during an immune challenge.

RNA-seq analysis of LPS-induced immune priming in silkworms (Bombyx mori) and the role of cytochrome P450 detoxification system in the process

While immune priming has been identified in many invertebrates, the intricate mechanisms that drive this process in insects continue to be a subject of mystery. In this study, we exposed silkworm larvae to varying doses of lipopolysaccharide (LPS) to induce immune priming and assessed their survival upon challenge with Bacillus thuringiensis (Bt). Transcriptome analysis was performed to identify differentially expressed genes (DEGs) associated with immune priming. The role of CYP450 genes in this process was further explored using RNA interference (RNAi) to knockdown CYP9E2 and CYP6K1, followed by measurements of detoxification enzyme activities and reactive oxygen species (ROS) levels. We found that LPS exposure significantly increased silkworm survival rates upon Bt challenge, indicating the induction of immune priming. Transcriptome analysis revealed 549 DEGs, including a large number involved in detoxification, immunity, and metabolism, suggesting a complex regulatory network that encompasses immune responses and metabolic pathways. Functional enrichment and gene set enrichment analysis (GSEA) highlighted the activation of immune signaling pathways and the involvement of detoxification processes. Knockdown of CYP9E2 and CYP6K1 resulted in increased ROS levels, decreased detoxification enzyme activities, and reduced survival rates post-Bt challenge, implicating the critical role of these genes in immune priming and detoxification. Our findings demonstrate that LPS-induced immune priming in silkworms involves the upregulation of CYP450 genes, which play a critical role in detoxification and immune response modulation. The study provides insights into the molecular mechanisms of immune priming in insects and highlights the potential of CYP9E2 and CYP6K1 as targets for enhancing disease resistance and pest management in insects.

The anti-inflammatory effect of lycopene on neutrophil infiltration in the spleen of mice exposed to Lipopolysaccharide (LPS)

Introduction: Lipopolysaccharide (LPS) is a major component of the outer membrane of gram-negative bacteria that can trigger inflammatory and oxidative stress responses. The purpose of this study is to observe the anti-inflammatory effect of lycopene in reducing neutrophil infiltration caused by LPS exposure in mice (Mus musculus). Objective: This laboratory experimental study used 25 male mice aged 3 months with body weights of ± 25 g – 35 g, divided into five groups with five replicates each. C(-) is the control group: no LPS or lycopene was administered. C(+): received LPS 0.042 mg/kg without lycopene. P1: received LPS 0.042 mg/kg and lycopene 0.3 mg/kg. P2: received LPS 0.042 mg/kg and lycopene 0.6 mg/kg. P3: received LPS 0.042 mg/kg and lycopene 0.9 mg/kg. LPS was given intraperitoneally on days 1 and 8, and lycopene was administered daily for 14 days. The spleen is located near the liver. After the organ was removed, it was placed in 10% formalin buffer for HE staining, and data were analyzed using the Kruskall Wallis test followed by the Mann-Whitney test (p&lt;0.05). Results: LPS exposure significantly increased the number of neutrophils (p&lt;0.05), and lycopene administration significantly reduced the number of neutrophils (p&lt;0.05) to normal levels similar to the control (C-) at all doses/groups. Conclusion: LPS exposure was proven to increase the number of neutrophils, and lycopene administration was able to reduce the number of neutrophils to normal levels in all dose groups.

Efficacy of lycopene in reducing malondialdehyde (MDA) levels due to Lipopolysaccharide (LPS) exposure in Mice (Mus musculus)

Objective: Lipopolysaccharide (LPS) is a major component of the outer membrane of gram-negative bacteria and can trigger inflammatory and oxidative stress responses. This study aims to observe the efficacy of lycopene in reducing malondialdehyde (MDA) levels due to lipopolysaccharide (LPS) exposure in mice (Mus musculus). Method: This laboratory experimental study used 25 male mice aged 3 months with body weights of ± 25 g – 35 g, divided into five groups with five replicates each. K(-) was the control group: no LPS or lycopene was administered. K(+): received LPS 0.042 mg/kg without lycopene. P1: received LPS 0.042 mg/kg and lycopene 0.3 mg/kg. P2: received LPS 0.042 mg/kg and lycopene 0.6 mg/kg. P3: received LPS 0.042 mg/kg and lycopene 0.9 mg/kg. LPS was given on days 1 and 8 intraperitoneally, and lycopene was administered daily for 14 days. MDA levels were measured using the ELISA method, and data analysis was performed using ANOVA followed by Tukey's Test (p&lt;0.05). Result: LPS exposure significantly increased MDA levels (p&lt;0.05), and lycopene administration significantly reduced MDA levels (p&lt;0.05) to normal levels similar to the control (-). Conclusion: LPS exposure significantly increased MDA levels, and lycopene administration was able to reduce MDA levels to normal limits.