Gut Microbiota and NLRP3 Inflammasome Activation in Hemodialysis Patients: Exploring the Link with Systemic Inflammation

Gut Microbiota and NLRP3 Inflammasome Activation in Hemodialysis Patients: Exploring the Link with Systemic Inflammation | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Gut Microbiota and NLRP3 Inflammasome Activation in Hemodialysis Patients: Exploring the Link with Systemic Inflammation Denise Mafra, Livia Alvarenga, Ludmila Cardozo, Júnia Schultz, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6248246/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 19 May, 2025 Read the published version in Molecular Biology Reports → Version 1 posted 7 You are reading this latest preprint version Abstract Background The Nod-Like Receptor Pyrin domain-containing 3 (NLRP3) inflammasome is a critical sensor of bacterial signals and metabolites, initiating an inflammatory response. Chronic kidney disease (CKD) is often accompanied by systemic inflammation, which can be involved with gut dysbiosis. Considering this interplay, we aimed to explore the potential association between NLRP3 inflammasome expression and gut microbiota in CKD patients undergoing hemodialysis (HD). Methods and results This research comprises a cross-sectional pilot study involving twelve HD patients [59.2 ± 13.4 years, six women, BMI 26.6 ± 3.5 kg/m 2 , 48.6 (20.1–77.2) months on dialysis]. The gut microbiota was evaluated by the 16S ribosomal RNA gene. The mRNA expression of NLRP3 was assessed using real-time quantitative polymerase chain reaction (qPCR). Plasma levels of IL-1β were measured by ELISA. A positive correlation between mRNA expression of NLRP3 and Lentisphaerae phylum and with Erysipelaloclostrium and Victivallis genus (p < 0.05) was observed. The IL-1β mRNA expression was positively correlated with Lentisphaerae and Spirochaetes phylum. Also, there was a positive correlation with the relative abundance of the genera Erysipelaloclostrium, Absicoccus, Colidextribacter, Desulfovibrio, Fournierella, Lawsonibacter, Ruminococcus , and Victivallis. Regarding Archaea, IL-1β mRNA expression was positively correlated with Methanobrevibacter. Conclusion In CDK patients undergoing hemodialysis, gut microbiota may be involved in NLRP3 activation and IL-1β expression, contributing to inflammation. NLRP3 inflammasome IL-1β gut microbiota dysbiosis hemodialysis Figures Figure 1 Figure 2 Figure 3 Introduction The interest in the instigating role of gut microbiota in maintaining host homeostasis and regulating the immune system has driven several studies [ 1 , 2 ]. In nephrology, preclinical studies since the 1990s have demonstrated the effects of uremia on intestinal permeability, gut microbiota, and endotoxemia in patients with chronic kidney disease (CKD) [ 3 , 4 ]. Since then, scientific and technological advances have deepened our understanding of the relationship between gut microbiota and CKD. Metagenome-wide analyses of two cohorts of hemodialysis CKD patients revealed alterations in the gut microbiome, characterized by 348 differentially abundant species. These included increased levels of Blautia spp ., Dorea spp., Eggerthellaceae , and decreased Prevotella and Roseburia species [ 5 ]. Key toxin-contributing species were identified, and microbial signatures associated with hemodialysis CKD patients correlated with disease progression in non-dialyzed CKD patients [ 5 ]. Gut dysbiosis – marked by perturbations in composition, diversity, and function of the gut microbiota – is now recognized as a determinant factor disrupting the host’s homeostatic relationship. More recently, an association between gut dysbiosis and increased mortality in hemodialysis patients has been reported [ 6 ]. Evidence suggests that unbalanced gut microbiota disrupts intestinal homeostasis through interactions with the gut-associated lymphoid tissue (GALT), exposing the host microorganisms and their metabolites. This exposure triggers systemic inflammation, a key mediator linking gut dysbiosis to adverse outcomes in CKD [ 6 – 8 ]. Several inflammatory pathways contribute to CKD pathology, including the complex cytoplasmatic proteins belonging to the innate immune system called inflammasomes. Among these, the Nod-Like Receptor Pyrin domain containing 3 (NLRP3) inflammasome plays a pivotal role in the inflammation upon receiving signals from bacteria and their metabolites, such as uremic toxins. Through pattern recognition receptors (PRRs), cells from the innate immune system recognize damage-associated molecular patterns (DAMPs) and pathogen-associated molecular patterns (PAMPs), such as lipopolysaccharide (LPS), peptidoglycan, and bacterial RNA. These signals induce NIMA-related kinase 7 (NEK7) to bind with NLRP3 and initiate inflammasome assembly [ 8 , 9 ]. The activation of the NLRP3 inflammasome results in the conversion of procaspase-1 into its active form, caspase-1. As an effector protein of the NLRP3 inflammasome, caspase-1 processes pro-IL-1β and pro-IL-18 into their active forms, IL-1β and IL-18, respectively, and also cleaves gasdermin D (GSDMD), into its active form, which then drives GSDMD-mediated pyroptosis and cell damage [ 9 ]. Dysregulated NLRP3 inflammasome activity in response to microbial components increases epithelial permeability and interleukin production, particularly IL-1β. Elevated IL-1β levels amplify systemic inflammation, stimulate downstream cytokine release, promote kidney inflammation, and drive Th17 cell differentiation and IL‑17 production. Excessive IL-1β secretion can lead to tissue damage and widespread inflammatory responses, posing significant risks to the host [ 10 ]. Preclinical studies have reinforced the hypothesis that gut microbiota interact with the NLRP3 inflammasome to activate it [ 11 , 12 ]. While research has focused mainly on bacterial species contributing to inflammation, archaea also represent a stable yet underexplored component of the human gut microbiota[ 13 ]. In mice, the microplastic intervention (environmental contaminants) negatively affected the gut microbiota, lowering the relative abundances of Dubosoella and increasing Bacteroides, Clostridia, Desulfovibrio, Enterorhabdus , and Gemella . This disruption of gut microbiota was linked to NLRP3 activation in the liver [ 11 ]. Similarly, Hong et al. (2024) reported that treatment with Ruminococcus gnavus in KK-Ay mice for eight weeks disrupted gut microbiota homeostasis, altered the expression of tight junction proteins, and impaired kidney function. Additionally, this intervention led to increased expression of NLRP3, contributing to an inflammatory state [ 12 ]. Additionally, studies on human proximal tubule epithelial cells have demonstrated that exposure to the uremic toxin indoxyl sulfate (IS) induces NLRP3 inflammasome activation, leading to increased expression of NLRP3, caspase-1, and IL-1β, as well as enhanced IL-1β secretion and caspase-1 activity. Furthermore, IS prompted the production of intracellular reactive oxygen species. In the same study, the authors observed a rising trend in the expression of the NLRP3 inflammasome in the CKD animal model, correlated with the plasma levels of IS, kynurenic acid, and hippuric acid [ 14 ]. Although there is increasing evidence supporting a connection between gut microbiota and NLRP3 inflammasome responses, data on CKD patients remain limited. This pilot study assesses the potential relationship between NLRP3 inflammasome activation, IL-1β expression, and gut microbiota composition in CKD patients undergoing hemodialysis. Materials and Methods Study design and participants This cross-sectional pilot study included hemodialysis patients, male and female, ≥ 18 years old, with arteriovenous fistula for vascular access in the upper limb, at least 6 months on dialysis. Patients with inflammatory diseases, cancer, AIDS, autoimmune disease, smokers, pregnancy, and patients using catabolic drugs, antioxidant vitamin supplements, pre-, pro, and symbiotic, and antibiotics in the last 3 months before the start of this study were excluded. Dialysis duration was 3-4.5 hours per session, three times weekly, with a blood flow > 250mL/min and a dialysate flow of 500mL/min. The Ethics Committee of Medicine Faculty of Federal Fluminense University reviewed and approved the study protocol (CAAE 47703315.6.0000.5243). All the patients who agreed to participate in the study signed an informed consent form. Body mass index (BMI) was calculated by dividing dry body weight (kg) by height squared (m). The kinetic index of dialysis (Kt/V) quantifies hemodialysis treatment and was calculated using the single-pool Daugirdas formula, with a reference value of > 1.2 according to NKF-KDOQI (National Kidney Foundation Kidney Disease Outcomes Quality Initiatives) guidelines. A dietary 24-hour recall for 3 days (including a dialysis day, a non-dialysis day, and a weekend day) was used to compute the average dietary intake, utilizing nutritional software (Nutwin, developed by the Department of Nutrition at the Federal University of São Paulo, UNIFESP). Demographic and clinical data were obtained by analyzing medical records and conducting interviews. Blood Sample Collection and Biochemical Analysis Blood samples were drawn before a regular hemodialysis session. Venous blood samples were drawn into an EDTA syringe (1.0 mg/mL). The blood was centrifuged (3,500 rpm for 15 minutes, at 4ºC) to obtain plasma, distributed in 1.5mL polypropylene Eppendorf ® tubes, separated into aliquots, and stored at -80°C for further analysis. An aliquot was used for whole blood analysis to obtain peripheral blood mononuclear cells (PBMCs), according to Cardozo et al. (2016)[ 15 ]. Routine biochemical analysis was performed using BioClin (Bio-clin BS-120 Chemistry Analyze). NLRP3 inflammasome and IL-1β analysis The mRNA expression of NLRP3 and IL-1β was assessed from PBMCs using real-time quantitative polymerase chain reaction (qPCR). PBMCs were isolated from blood, and RNA was extracted using the SV Total RNA Isolation System (Promega®). cDNA was synthesized using the High-Capacity cDNA Reverse Transcription kit (Thermo Fisher®). A TaqMan Gene Expression assay (Thermo Fisher®) was performed to detect the expression of NLRP3 mRNA (Hs00918082_m1), IL-1β mRNA (Hs00918082_m1) and the control gene GAPDH (Hs02758991_g1). The ABI Prism 7500 Sequence Detection System (Applied Biosystems®) and standard cycling conditions were used for PCR amplification. The expression level was calculated using the ΔΔCT (delta cycle threshold) method. Gut microbiota analysis The patients received a sterile stool collection tube and instructions for collecting stool samples. The collected stool samples were stored at -20°C until further processing. According to the manufacturer's protocol, DNA extraction was performed using the Quick-DNA Fecal/Soil Microbe Miniprep Kit (Zymo Research). The concentration and quality of the total DNA were determined using a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). The V4 region of the 16S rRNA gene was sequenced for the gut microbiota analysis. Firstly, the region was amplified via PCR using the primers 515F (5’-GTGYCAGCMGCCGCGGTAA-3’) and 806R (5’-GGACTACNVGGGTWTCTAAT-3’), with universal Illumina adapters. The thermocycling conditions included an initial denaturation at 94°C for 3 minutes, followed by 32 cycles of 94°C for 45 seconds, 50°C for 1 minute, and 72°C for 90 seconds, with a final extension at 72°C for 10 minutes. The resulting amplicons were barcoded, pooled, and sequenced on the Illumina NovaSeq PE250 platform (targeting 0.1 million raw reads per sample) according to the manufacturer’s protocol at Novogene (California, USA). A total of 12 samples were subjected to amplicon sequencing. The raw 16S rRNA gene sequencing reads were processed using the USEARCH pipeline (v. 11). Sequence abundances were determined, and Operational Taxonomic Units (OTUs) were clustered at 97% identity. Chimeric sequences were filtered through a denoising step to generate a Zero-radius OTU (zOTU) table. Feature and taxonomy tables, along with metadata, were exported as ‘phyloseq’ objects for further analysis in R (v.4.1.2). For the correlation assessment, the data was then transformed using centered log ratio (CLR) transformation [ 16 ] and submitted to Spearman’s rank correlation coefficient, with a significance threshold of p < 0.05) using the ‘corrplot’ R package. All plots and graphs were generated with the R package ‘ggplot2’ v.4.1.2. Statistical Analyses Data were expressed as median and quartile intervals or mean and SD as appropriate. Statistical significance was accepted as p < 0.05. Statistical analyses were performed using the statistical software SPSS 24.0. The Wilk test verified the distribution of descriptive variables in the present study. According to the distribution of explanatory variables, the results are expressed as mean ± SD (standard deviation) or median (quartile intervals). Statistical analyses were performed using SPSS 24.0 (SPSS, Inc., Chicago, IL, USA). Results The study involved 12 CKD patients undergoing hemodialysis. Table 1 presents the general, biochemical, inflammatory, and dietary characteristics of HD patients. Hypertensive nephrosclerosis was the primary etiological factor of CKD. According to BMI classification, 50% of the patients were overweight, and none were malnourished. All patients showed adequate Kt/V. Table 1 The HD patients' general, biochemical, inflammatory, and dietary characteristics. Parameters Overall (n = 12) General features Female/Male (%) 50/50 Age (years) 59.2 ± 13.4 Kt/V 1.4 ± 0.1 HD vintage (months) 48.6 (20.1–77.2) Body mass index (kg/m²) 26.6 ± 3.5 Transcription factors mRNA NLPR3 1.1 ± 0.4 mRNA Interleukin 1-beta 1.4 (0.8–2.7) Routine Biochemical Total Cholesterol (mg/dL) 143.3 ± 33.6 Triglycerides (mg/dL) 119 (93.6-169.1) High-Density Lipoprotein (mg/dL) 42.5 ± 11.2 Low-Density Lipoprotein (mg/dL) 73.6 ± 26.3 Calcium (mg/dL) 8.8 ± 0.5 Phosphorus (mg/dL) 5.3 ± 0.9 Albumin (mg/dL) 3.6 ± 0.2 Potassium (mg/dL) 4.8 ± 0.2 Urea (mg/dL) 148.7 ± 34.5 Hemoglobin (mg/dL) 11.9 ± 1.4 Parathormone (mg/dL) 517 (381–1199) high-sensitivity C-reactive protein (mg/L) 2.5 (0.2–9.2) Dietary assessments Energy (Kcal/day) 1500 ± 479 Carbohydrate (g/day) 214 ± 66.8 Protein (g/day) 70 ± 22.8 Lipid (g/day) 42 ± 18.6 Phosphorus (mg/day) 1006 ± 343 Potassium (mg/day) 1904 ± 639 Non-normally distributed variables are presented as median (25th percentile and 75th percentile) and normally distributed variables as mean ± SD. Kt/V: kinetic index of dialysis; NLPR3: Nod-Like Receptor Pyrin domain containing 3 Regarding the gut microbiota and its relationship with inflammation markers, Fig. 1 indicates that the abundance of Fusobacteria phylum is negatively correlated (p < 0.05) with the mRNA expression levels of NLRP3 and IL-1β. In contrast, members of the phylum Lentisphaerae exhibited a positive correlation with the mRNA expression levels of NLRP3 and IL-1β (p < 0.05). Additionally, a positive correlation was observed between IL-1β and the phylum Spirochaetes. As expected, NLRP3 and IL-1β expression levels were positively correlated (p < 0.05). Figure 2 illustrates the correlation between gut microbial genera and inflammatory markers. The relative abundance of Fusobacterium was negatively correlated with the mRNA expression levels of NLRP3 and IL-1β (p < 0.05). Additionally, the relative abundance of Erysipelatoclostridium and Victivallis positively correlates with the mRNA expression level of NLRP3 (p < 0.05). Furthermore, the relative abundances of Abscicoccus, Colidextribacter, Desulfovibrio, Fournierella, Lawsonibacter, Ruminococcus , and Victivallis were positively correlated with IL-1β mRNA expression (p < 0.05). Figure 3 highlights the significant positive correlation between IL-1β expression and the archaeal genus Methanosphaera in CKD patients undergoing hemodialysis. Discussion This study investigated the association between gut microbiota and the NLRP3 inflammasome in patients with CKD undergoing hemodialysis. Our results suggest that the Lentisphaerae and Spirochaetes phyla, as well as members of the genera Erysipelatoclostridium , Victivallis , Abscoccus , Colidextribacter , Desulfovibrio , Fournierella , Lawsonibacter , and Ruminococcus may play a role in activating the NLRP3 inflammasome and IL-1β expression. Also, Methanosphaera , a genus of Archaea, was positively correlated with IL-1β expression, potentially contributing to the inflammation linked to gut dysbiosis in these patients. Identifying the constituent members of the gut microbiota may help us understand the structure and function of the gut microbial community characteristic of specific conditions. In this study, we observed that the Lentisphaerae phylum was positively correlated with NLRP3 and IL-1β. Lentisphaerae is a phylum associated with ischemic stroke lesions [ 17 ] and potentially linked to myasthenia gravis [ 18 ]. One study showed that after one year of lifestyle intervention involving the Mediterranean diet, individuals with nonalcoholic fatty liver disease (NAFLD) demonstrated a decrease in the Lentisphaerae phylum, suggesting that this genus may be involved in inflammatory pathways [ 19 ]. Victivallis belongs to the phylum Lentisphaerae , which was positively associated with NLRP3 inflammasome and IL-1β expression in this study. Prior research indicated that Victivallis may be related to inflammatory factors in acute respiratory distress syndrome [ 20 ]. Victivallis levels were significantly higher in individuals with colorectal cancer compared to healthy controls [ 21 ]. Furthermore, a Mendelian randomization analysis by Miao et al. (2024) demonstrated that the genus Victivallis had a causal relationship with hypertension[ 22 ]. Our study also suggests a potential role for Spirochaetes in the expression of IL-1β. Spirochaetes play an important role in the polymicrobial infections that lead to periodontitis. Oral spirochaetes have been shown to activate both innate and adaptive immune responses. Buford et al. (2018) found that the relative abundance of Spirochaetes was significantly lower in older adults compared to healthy young individuals. However, a positive correlation exists between Spirochaetes and Insulin-like growth factor-1 (IGF-1). It is important to note that the phylum is a broad taxonomic category that includes a diverse range of microorganisms with various functions and roles [ 23 ]. Absicoccus , part of the Erysipelotrichaceae family, was described in 2020 [ 24 , 25 ], and its role in inflammation remains unclear. However, the Erysipelotrichaceae family is associated with the metabolism of L-carnitine from dietary sources, resulting in increased circulating levels of trimethylamine N-oxide (TMAO) and heightened levels of inflammatory cytokines such as IL-1 β, IL-6, tumor necrosis factor-alpha (TNF-α), and TNF-β [ 5 , 26 ]. Additionally, exposure to pollutants like lead has been shown to affect the abundance of pathogenic bacterial families, including Erysipelotrichaceae. This is linked to increased levels of IL-8, IL-10, IL-1, and TNF-α. These findings provide valuable insights for further investigation into the role of gender in inflammation and chronic non-communicable diseases, including CKD [ 27 ]. The Colidextribacter genus was recently isolated from the human right colon. However, little is known about its function in metabolic pathways associated with human health [ 28 ]. A Crohn's and Colitis Canada Genetic Environmental Microbial (CCC-GEM) cohort showed that individuals with impaired barrier function had an increased abundance of Colidextribacter , among other taxonomic changes [ 29 ]. In an experimental model of liver fibrosis, Colidextribacter was identified as a harmful bacteria associated with damage to the intestinal barrier [ 30 ]. Contrary to our findings, a study in rats with NAFLD found a negative correlation between Colidextribacter and serum TNF-α [ 31 ]. Another study in rats with hyperuricemia also identified Colidextribacter as a beneficial bacterium that increases with marine fish-derived peptide supplementation, alongside a decrease in inflammatory factors (TNF-α, IL-6, and IL-10) [ 32 ]. Finally, an experimental study involving Shen-Ling-Bai-Zhu-San, a well-known formulation in traditional Chinese medicine, showed that its use inhibits NLRP3, IL-1β, and IL-18 expression levels. This finding is linked to restoring the dysfunctional intestinal microbiota in colitis-affected mice, which includes an increase in the abundance of bacteria that produce short-chain fatty acids, such as Colidextribacter [ 33 ]. Contradictions persist in the literature regarding the harmful or beneficial roles of the Colidextribacter genus. The genus Desulfovibrio consists of sulfate-reducing bacteria that inhabit the human intestine, with their numbers increasing under conditions linked to microbial dysbiosis and inflammation [ 34 ]. A greater abundance of Desulfovibrio correlates with increased intestinal epithelial barrier permeability [ 35 ]. Additionally, caspase-1, IL-1β, and IL-18 expressions also elevate Desulfovibrio abundance. Desulfovibrio flagellins trigger the inflammatory response and the release of cytokines and caspases through the activation of nucleotide-binding oligomerization domain-like receptor (NLR) family of apoptosis inhibitory proteins (NAIP) / NLR family caspase activation and recruitment domain-containing proteins 4 (NLRC4) inflammasome, which contributes to intestinal damage[ 36 ]. Moreover, a study in rats indicated intestinal modulation by lycopene, evidenced by a decrease in hepatic protein expressions of NLRP3, Pro-Caspase-1, Caspase-1, and factor nuclear kappa B (NF-ΚB), which is associated with lower fecal levels of Desulfovibrio [ 37 ]. Wang et al. (2022) reported that Fournierella contributes to worsening intestinal inflammation by disrupting bile secretion and tryptophan metabolism, which affects intestinal permeability and results in diarrhea in rabbits[ 38 ]. However, other studies present contradictory findings, as calves suffering from damp-heat diarrhea, a common condition in Chinese dairy farms, showed a decrease in the abundance of the genus Fournierella [ 39 ]. Lawsonibacter is a Gram-positive, anaerobic, non-motile, non-spore-forming, and asaccharolytic bacterium. One study examined the changes in fecal microbiota following Roux-en-Y gastric bypass in obese patients. The results indicated improved patient health, reduced inflammation, and notable alterations in the intestinal microbiome, including a decrease in the Lawsonibacter genus, among other genera [ 40 ]. In contrast, a diet rich in whole corn grains for seven weeks enhanced lambs’ microbiota by increasing the Lawsonibacter genus's relative abundance in the colonic mucosa [ 41 ]. Fecal microbiota transplantation was examined in irritable bowel syndrome (IBS) patients. In addition to reducing bloating and severity, bacteria such as Lawsonibacter were enriched by week 12 [ 42 ]. Ruminococcus species are strictly anaerobic, gram-positive, and non-motile cocci that do not produce endospores, and it is essential to the microbial communities of ruminants and humans. They require fermentable carbohydrates for their growth and are a critical source of short-chain fatty acid (SCFA) production [ 43 ]. Profiles of Ruminococcus bromii in healthy individuals showed increased abundance when consuming diets rich in resistant starch, indicating that these bacteria may be vital for the fermentation of complex carbohydrates in the large intestine [ 44 ]. In rats with ulcerative colitis that were subjected to Kui jie tong (KJT), a herbal from traditional Chinese medicine, it was observed that KJT inhibited the expression of NLRP3, caspase-1, and cleaved-caspase-1 mRNA and protein in colon tissues. Furthermore, KJT reduced the expression of IL-1β, IL-18, and IL-33 levels in colon tissue and serum, and added to these findings, it also increased unclassified_g__Ruminococcus_1 levels [ 45 ]. However, enrichment of Lachnospiraceae in allergic children with overgrowth of Ruminococcus gnavus was associated with respiratory allergies. In the same study, fed mice with purified R. gnavus developed histological evidence of airway inflammation and stimulated cytokine secretion [ 46 ]. In a model of lipopolysaccharide-induced diarrhea in mice, Ruminococcus showed a positive correlation with inflammatory cytokines. It was negatively correlated with tight junction proteins and the expressions of Caspase-1 and NLRP3 [ 47 ]. Oral administration of R. gnavus adversely affected the kidneys in mice, leading to increased levels of nitrogen, creatinine, proteinuria, uremic toxins, and inflammatory markers (NLRP3 and IL-6) [ 12 ]. Methanosphaera is an archaeal genus recognized as a vital component of the human microbiome. These organisms play a key role in global carbon cycling due to their ability to produce methane, a potent greenhouse gas, as a byproduct of their energy production. It is believed that the composition of the gut methanogen community contributes to variations in methane emissions. Moreover, there is increasing evidence that methanogens and/or the methane they produce may significantly affect human health and disease [ 48 ]. The molecular interactions between archaea and the immune system and their potential role in inflammation remain poorly understood [ 49 ]. Notably, an increased abundance of Methanosphaera spp. has been found in individuals with inflammatory bowel disease [ 50 ]. Specifically, Bang et al. (2014) demonstrated that the archaeal species M. stadtmanae induced the release of IL-1β in monocyte-derived dendritic cells[ 51 ]. Additionally, M. stadtmanae was shown to activate the NLRP3 inflammasome in human monocytic BLaER1 knockout cells. More research is necessary to understand better the inflammatory responses triggered by archaea [ 48 ]. We acknowledge the limitations of this study. Due to the cross-sectional design, a causal relationship between gut microbiota and NLRP3 inflammasome activation could not be established. Factors such as other inflammatory biomarkers, metabolites, and diet may have influenced the observed associations. Additionally, the small sample size may limit the generalizability of the findings. Future studies should include power analysis, considering the NLRP3 inflammasome components investigated here. Conclusion In conclusion, gut microbiota components in hemodialysis patients may significantly activate the NLRP3 inflammasome. The interaction between specific gut microbiota taxa and NLRP3 activation could contribute to disrupting the gut barrier and systemic inflammation. However, further investigations are needed to clarify the precise mechanisms underlying the interactions between gut microbiota and the NLRP3 inflammasome in individuals with CKD. Our findings provide valuable insights into the mechanisms driving inflammation caused by gut dysbiosis in CKD. Abbreviations 16S rRNA – 16S ribosomal RNA BMI – Body Mass Index CCC-GEM – Crohn's and Colitis Canada Genetic Environmental Microbial CKD – Chronic Kidney Disease CLR – Centered Log Ratio DAMPs – Damage-Associated Molecular Patterns EDTA – Ethylenediaminetetraacetic Acid GAPDH – Glyceraldehyde-3-Phosphate Dehydrogenase GALT – Gut-Associated Lymphoid Tissue GSDMD – Gasdermin D HD – Hemodialysis IGF-1 – Insulin-like Growth Factor-1 IL-1 β – Interleukin-1 Beta IL-10 – Interleukin-10 IL-17 – Interleukin-17 IL-18 – Interleukin-18 IL-33 – Interleukin-33 IL-6 – Interleukin-6 IL-8 – Interleukin-8 IS – Indoxyl Sulfate Kt/V – Kinetic Index of Dialysis KJT – Kui jie tong NKF-KDOQI – National Kidney Foundation Kidney Disease Outcomes Quality Initiatives NIMA – Never In Mitosis A NEK7 – NIMA-related Kinase 7 NLRP3 – Nod-Like Receptor Pyrin Domain Containing 3 OTUs – Operational Taxonomic Units PBMCs – Peripheral Blood Mononuclear Cells PCR – Polymerase Chain Reaction PAMPs – Pathogen-Associated Molecular Patterns PRRs – Pattern Recognition Receptors qPCR – Quantitative Polymerase Chain Reaction SD – Standard Deviation SCFA – Short-Chain Fatty Acids SPSS – Statistical Package for the Social Sciences TMAO – Trimethylamine N-oxide TNF-α – Tumor Necrosis Factor-alpha TNF-β – Tumor Necrosis Factor-beta Th17 – T Helper 17 Cells zOTU – Zero-radius Operational Taxonomic Unit Declarations There are no conflicts of interest to declare. The study protocol was reviewed and approved by the institutional ethics committee. All patients who agreed to participate in the study signed an informed consent form. This work was supported by Prof. Alexandre Soares Rosado’s KAUST Baseline Grant (BAS/1/1096-01-01). Author Contribution D.M. and N.A.B. designed the research, L.A. and L.C. conducted the study and drafted the initial manuscript, J.S and A.S.R. conducted the experiments, D.M. and N.A.B. revised the manuscript. References Hou K, Wu ZX, Chen XY, Wang JQ, Zhang D, Xiao C et al (2022) Microbiota in health and diseases. Signal Transduct Target Ther [Internet]. [cited 2025 Feb 3];7. Available from: https://pubmed.ncbi.nlm.nih.gov/35461318/ Krukowski H, Valkenburg S, Madella AM, Garssen J, van Bergenhenegouwen J, Overbeek SA et al (2023) Gut microbiome studies in CKD: opportunities, pitfalls and therapeutic potential. Nat Rev Nephrol [Internet]. [cited 2025 Feb 3];19:87–101. Available from: https://pubmed.ncbi.nlm.nih.gov/36357577/ Hida M, Aiba Y, Sawamura S, Suzuki N, Satoh T, Koga Y (1996) Inhibition of the accumulation of uremic toxins in the blood and their precursors in the feces after oral administration of Lebenin, a lactic acid bacteria preparation, to uremic patients undergoing hemodialysis. Nephron [Internet]. [cited 2025 Feb 3];74:349–55. Available from: https://pubmed.ncbi.nlm.nih.gov/8893154/ Vaziri ND, Wong J, Pahl M, Piceno YM, Yuan J, Desantis TZ et al (2013) Chronic kidney disease alters intestinal microbial flora. Kidney Int [Internet]. [cited 2025 Feb 3];83:308–15. Available from: https://pubmed.ncbi.nlm.nih.gov/22992469/ Zhang P, Wang X, Li S, Cao X, Zou J, Fang Y et al (2023) Metagenome-wide analysis uncovers gut microbial signatures and implicates taxon-specific functions in end-stage renal disease. Genome Biol [Internet]. [cited 2025 Jan 28];24. Available from: https://pubmed.ncbi.nlm.nih.gov/37828586/ Lin TY, Wu PH, Lin YT, Hung SC Gut dysbiosis and mortality in hemodialysis patients. NPJ Biofilms Microbiomes [Internet]. 2021 [cited 2025 Jan 28];7. Available from: https://pubmed.ncbi.nlm.nih.gov/33658514/ Schroder K, Tschopp J (2010) The inflammasomes. Cell [Internet]. [cited 2025 Jan 28];140:821–32. Available from: https://pubmed.ncbi.nlm.nih.gov/20303873/ Zheng X, Wan J, Tan G (2023) The mechanisms of NLRP3 inflammasome/pyroptosis activation and their role in diabetic retinopathy. Front Immunol [Internet]. [cited 2025 Jan 28];14. Available from: https://pubmed.ncbi.nlm.nih.gov/37180116/ Liu F, Gao C Regulation of the Inflammasome Activation by Ubiquitination Machinery. Adv Exp Med Biol [Internet]. 2024 [cited 2025 Jan 28];1466:123–34. Available from: https://pubmed.ncbi.nlm.nih.gov/39546140/ Anders HJ Of Inflammasomes and Alarmins: IL-1β and IL-1α in Kidney Disease. J Am Soc Nephrol [Internet]. 2016 [cited 2025 Jan 28];27:2564–75. Available from: https://pubmed.ncbi.nlm.nih.gov/27516236/ Xu R, Cao J, wen, Lv H li, Geng Y, Guo Myao (2024) Polyethylene microplastics induced gut microbiota dysbiosis leading to liver injury via the TLR2/NF-κB/NLRP3 pathway in mice. Sci Total Environ [Internet]. [cited 2025 Jan 28];917. Available from: https://pubmed.ncbi.nlm.nih.gov/38286276/ Hong J, Fu T, Liu W, Du Y, Bu J, Wei G et al (2024) Specific Alternation of Gut Microbiota and the Role of Ruminococcus gnavus in the Development of Diabetic Nephropathy. J Microbiol Biotechnol [Internet]. [cited 2025 Jan 28];34:547–61. Available from: https://pubmed.ncbi.nlm.nih.gov/38346799/ Mohammadzadeh R, Mahnert A, Duller S, Moissl-Eichinger C (2022) Archaeal key-residents within the human microbiome: characteristics, interactions and involvement in health and disease. Curr Opin Microbiol [Internet]. [cited 2025 Jan 28];67. Available from: https://pubmed.ncbi.nlm.nih.gov/35427870/ Mihajlovic M, Krebber MM, Yang Y, Ahmed S, Lozovanu V, Andreeva D et al (2021) Protein-Bound Uremic Toxins Induce Reactive Oxygen Species-Dependent and Inflammasome-Mediated IL-1β Production in Kidney Proximal Tubule Cells. Biomedicines [Internet]. [cited 2025 Jan 28];9. Available from: https://pubmed.ncbi.nlm.nih.gov/34680443/ Cardozo LFMF, Stockler-Pinto MB, Mafra D (2016) Brazil nut consumption modulates Nrf2 expression in hemodialysis patients: A pilot study. Mol Nutr Food Res [Internet]. [cited 2025 Jan 28];60:1719–24. Available from: https://pubmed.ncbi.nlm.nih.gov/26992136/ Aitchison J The Statistical Analysis of Compositional Data. Journal of the Royal Statistical Society: Series B (Methodological) [Internet]. 1982 [cited 2025 Feb 3];44:139–60. Available from: https://onlinelibrary.wiley.com/doi/full/ 10.1111/j.2517-6161.1982.tb01195.x Zou X, Wang L, Xiao L, Wang S, Zhang L (2022) Gut microbes in cerebrovascular diseases: Gut flora imbalance, potential impact mechanisms and promising treatment strategies. Front Immunol [Internet]. [cited 2025 Jan 28];13. Available from: https://pubmed.ncbi.nlm.nih.gov/36389714/ Shi J, Yi M, Xie S, Wang Z, Zhang X, Tan X et al (2024) Mendelian randomization study revealed a gut microbiota-neuromuscular junction axis in myasthenia gravis. Sci Rep [Internet]. [cited 2025 Jan 28];14. Available from: https://pubmed.ncbi.nlm.nih.gov/38291090/ Gómez-Pérez AM, Ruiz-Limón P, Salas-Salvadó J, Vioque J, Corella D, Fitó M et al (2023) Gut microbiota in nonalcoholic fatty liver disease: a PREDIMED-Plus trial sub analysis. Gut Microbes [Internet]. [cited 2025 Feb 3];15. Available from: https://pubmed.ncbi.nlm.nih.gov/37345236/ Ma J, Zhu Z, Yishajiang Y, Alarjani KM, Hong L, Luo L Role of gut microbiota and inflammatory factors in acute respiratory distress syndrome: a Mendelian randomization analysis. Front Microbiol [Internet]. 2023 [cited 2025 Jan 28];14. Available from: https://pubmed.ncbi.nlm.nih.gov/38173678/ Sánchez-Alcoholado L, Ordóñez R, Otero A, Plaza-Andrade I, Laborda-Illanes A, Medina JA et al Gut Microbiota-Mediated Inflammation and Gut Permeability in Patients with Obesity and Colorectal Cancer. Int J Mol Sci [Internet]. 2020 [cited 2025 Jan 28];21:1–20. Available from: https://pubmed.ncbi.nlm.nih.gov/32947866/ Miao C, Xu X, Huang S, Kong L, He Z, Wang Y et al (2024) The Causality between Gut Microbiota and Hypertension and Hypertension-related Complications: A Bidirectional Two-Sample Mendelian Randomization Analysis. Hellenic J Cardiol [Internet]. [cited 2025 Jan 28]; Available from: https://pubmed.ncbi.nlm.nih.gov/38336261/ Buford TW, Carter CS, VanDerPol WJ, Chen D, Lefkowitz EJ, Eipers P et al Composition and richness of the serum microbiome differ by age and link to systemic inflammation. Geroscience [Internet]. 2018 [cited 2025 Jan 28];40:257–68. Available from: https://pubmed.ncbi.nlm.nih.gov/29869736/ Oren A, Göker M (2024) Notification that new names of prokaryotes, new combinations and new taxonomic opinions have appeared in volume 74, part 9 of the IJSEM. Int J Syst Evol Microbiol [Internet]. [cited 2025 Jan 28];74. Available from: https://pubmed.ncbi.nlm.nih.gov/39700045/ Shin Y, Paek J, Kim H, Kook JK, Kim JS, Kim SH et al (2020) Absicoccus porci gen. nov., sp. nov., a member of the family Erysipelotrichaceae isolated from pig faeces. Int J Syst Evol Microbiol [Internet]. [cited 2025 Jan 28];70:732–7. Available from: https://pubmed.ncbi.nlm.nih.gov/31702538/ Zhang X, Wu Q, Zhao Y, Yang X (2019) Decaisnea insignis Seed Oil Inhibits Trimethylamine- N-oxide Formation and Remodels Intestinal Microbiota to Alleviate Liver Dysfunction in l-Carnitine Feeding Mice. J Agric Food Chem [Internet]. [cited 2025 Jan 28];67:13082–92. Available from: https://pubmed.ncbi.nlm.nih.gov/31671940/ Liu H, Zhang H, Yu Q, Zhang S, Tu X, Zhuang F et al (2023) Lead induced structural and functional damage and microbiota dysbiosis in the intestine of crucian carp (Carassius auratus). Front Microbiol [Internet]. [cited 2025 Jan 28];14. Available from: https://pubmed.ncbi.nlm.nih.gov/37731918/ Ricaboni D, Mailhe M, Cadoret F, Vitton V, Fournier PE, Raoult D (2016) Colidextribacter massiliensis gen. nov., sp. nov., isolated from human right colon. New Microbes New Infect [Internet]. [cited 2025 Jan 28];17:27–9. Available from: https://pubmed.ncbi.nlm.nih.gov/28275436/ Leibovitzh H, Lee SH, Xue M, Raygoza Garay JA, Hernandez-Rocha C, Madsen KL et al Altered Gut Microbiome Composition and Function Are Associated With Gut Barrier Dysfunction in Healthy Relatives of Patients With Crohn’s Disease. Gastroenterology [Internet]. 2022 [cited 2025 Jan 28];163:1364–1376.e10. Available from: https://pubmed.ncbi.nlm.nih.gov/35850197/ He X, Liang J, Li X, Wang Y, Zhang X, Chen D et al (2024) Dahuang zhechong pill ameliorates hepatic fibrosis by regulating gut microbiota and metabolites. J Ethnopharmacol [Internet]. [cited 2025 Jan 28];321. Available from: https://pubmed.ncbi.nlm.nih.gov/37967779/ Liu X, Zhang Y, Li W, Zhang B, Yin J, Liuqi S et al (2022) Fucoidan Ameliorated Dextran Sulfate Sodium-Induced Ulcerative Colitis by Modulating Gut Microbiota and Bile Acid Metabolism. J Agric Food Chem [Internet]. [cited 2025 Jan 28];70:14864–76. Available from: https://pubmed.ncbi.nlm.nih.gov/36378195/ Wu C, Hu Q, Peng X, Luo J, Zhang G (2023) Marine Fish Protein Peptide Regulating Potassium Oxonate-Induced Intestinal Dysfunction in Hyperuricemia Rats Helps Alleviate Kidney Inflammation. J Agric Food Chem [Internet]. [cited 2025 Jan 28];71:320–30. Available from: https://pubmed.ncbi.nlm.nih.gov/36530149/ Gao Q, Tian W, Yang H, Hu H, Zheng J, Yao X et al (2024) Shen-Ling-Bai-Zhu-San alleviates the imbalance of intestinal homeostasis in dextran sodium sulfate-induced colitis mice by regulating gut microbiota and inhibiting the NLRP3 inflammasome activation. J Ethnopharmacol [Internet]. [cited 2025 Jan 28];319. Available from: https://pubmed.ncbi.nlm.nih.gov/37704122/ Singh SB, Coffman CN, Carroll-Portillo A, Varga MG, Lin HC (2021) Notch Signaling Pathway Is Activated by Sulfate Reducing Bacteria. Front Cell Infect Microbiol [Internet]. [cited 2025 Jan 28];11. Available from: https://pubmed.ncbi.nlm.nih.gov/34336718/ Singh SB, Coffman CN, Varga MG, Carroll-Portillo A, Braun CA, Lin HC (2022) Intestinal Alkaline Phosphatase Prevents Sulfate Reducing Bacteria-Induced Increased Tight Junction Permeability by Inhibiting Snail Pathway. Front Cell Infect Microbiol [Internet]. [cited 2025 Jan 28];12. Available from: https://pubmed.ncbi.nlm.nih.gov/35694541/ An Y, Zhai Z, Wang X, Ding Y, He L, Li L et al (2023) Targeting Desulfovibrio vulgaris flagellin-induced NAIP/NLRC4 inflammasome activation in macrophages attenuates ulcerative colitis. J Adv Res [Internet]. [cited 2025 Jan 28];52:219–32. Available from: https://pubmed.ncbi.nlm.nih.gov/37586642/ Gao X, Zhao X, Liu M, Zhao H, Sun Y Lycopene prevents non-alcoholic fatty liver disease through regulating hepatic NF-κB/NLRP3 inflammasome pathway and intestinal microbiota in mice fed with high-fat and high-fructose diet. Front Nutr [Internet]. 2023 [cited 2025 Jan 28];10. Available from: https://pubmed.ncbi.nlm.nih.gov/37032779/ Wang J, Zhao K, Kang Z, Wang M, Chen Y, Fan H et al The Multi-Omics Analysis Revealed a Metabolic Regulatory System of Cecum in Rabbit with Diarrhea. Animals (Basel) [Internet]. 2022 [cited 2025 Jan 28];12. Available from: https://pubmed.ncbi.nlm.nih.gov/35565618/ Yan Z, Zhang K, Zhang K, Wang G, Wang L, Zhang J et al (2022) Integrated 16S rDNA Gene Sequencing and Untargeted Metabolomics Analyses to Investigate the Gut Microbial Composition and Plasma Metabolic Phenotype in Calves With Dampness-Heat Diarrhea. Front Vet Sci [Internet]. [cited 2025 Jan 28];9. Available from: https://pubmed.ncbi.nlm.nih.gov/35242833/ Prykhodko O, Burleigh S, Campanello M, Iresjö BM, Zilling T, Ljungh Å et al Long-Term Changes to the Microbiome, Blood Lipid Profiles and IL-6 in Female and Male Swedish Patients in Response to Bariatric Roux-en-Y Gastric Bypass. Nutrients [Internet]. 2024 [cited 2025 Jan 28];16. Available from: https://pubmed.ncbi.nlm.nih.gov/38398821/ Cheng C, Yin Y, Bian G (2022) Effects of whole maize high-grain diet feeding on colonic fermentation and bacterial community in weaned lambs. Front Microbiol [Internet]. [cited 2025 Jan 28];13. Available from: https://pubmed.ncbi.nlm.nih.gov/36569065/ Yau YK, Su Q, Xu Z, Tang W, Ching JYL, Mak JWY et al (2023) Randomised clinical trial: Faecal microbiota transplantation for irritable bowel syndrome with diarrhoea. Aliment Pharmacol Ther [Internet]. [cited 2025 Jan 28];58:795–804. Available from: https://pubmed.ncbi.nlm.nih.gov/37667968/ Rainey FA, Janssen PH Phylogenetic analysis by 16S ribosomal DNA sequence comparison reveals two unrelated groups of species within the genus Ruminococcus. FEMS Microbiol Lett [Internet]. 1995 [cited 2025 Jan 28];129:69–73. Available from: https://pubmed.ncbi.nlm.nih.gov/7781992/ Abell GCJ, Cooke CM, Bennett CN, Conlon MA, McOrist AL Phylotypes related to Ruminococcus bromii are abundant in the large bowel of humans and increase in response to a diet high in resistant starch. FEMS Microbiol Ecol [Internet]. 2008 [cited 2025 Jan 28];66:505–15. Available from: https://pubmed.ncbi.nlm.nih.gov/18616586/ Xue S, Xue Y, Dou D, Wu H, Zhang P, Gao Y et al Kui Jie Tong Ameliorates Ulcerative Colitis by Regulating Gut Microbiota and NLRP3/Caspase-1 Classical Pyroptosis Signaling Pathway. Dis Markers [Internet]. 2022 [cited 2025 Jan 28];2022. Available from: https://pubmed.ncbi.nlm.nih.gov/35832643/ Chua HH, Chou HC, Tung YL, Chiang BL, Liao CC, Liu HH et al Intestinal Dysbiosis Featuring Abundance of Ruminococcus gnavus Associates With Allergic Diseases in Infants. Gastroenterology [Internet]. 2018 [cited 2025 Jan 28];154:154–67. Available from: https://pubmed.ncbi.nlm.nih.gov/28912020/ Chen X, Kong Q, Zhao X, Zhao C, Hao P, Irshad I et al Sodium acetate/sodium butyrate alleviates lipopolysaccharide-induced diarrhea in mice via regulating the gut microbiota, inflammatory cytokines, antioxidant levels, and NLRP3/Caspase-1 signaling. Front Microbiol [Internet]. 2022 [cited 2025 Jan 28];13. Available from: https://pubmed.ncbi.nlm.nih.gov/36386709/ Volmer JG, McRae H, Morrison M (2023) The evolving role of methanogenic archaea in mammalian microbiomes. Front Microbiol [Internet]. [cited 2025 Jan 28];14. Available from: https://pubmed.ncbi.nlm.nih.gov/37727289/ Mohammadzadeh R, Mahnert A, Duller S, Moissl-Eichinger C (2022) Archaeal key-residents within the human microbiome: characteristics, interactions and involvement in health and disease. Curr Opin Microbiol [Internet]. [cited 2025 Jan 28];67. Available from: https://pubmed.ncbi.nlm.nih.gov/35427870/ Lecours PB, Marsolais D, Cormier Y, Berberi M, Haché C, Bourdages R et al (2014) Increased prevalence of Methanosphaera stadtmanae in inflammatory bowel diseases. PLoS One [Internet]. [cited 2025 Jan 28];9. Available from: https://pubmed.ncbi.nlm.nih.gov/24498365/ Bang C, Weidenbach K, Gutsmann T, Heine H, Schmitz RA (2014) The intestinal archaea Methanosphaera stadtmanae and Methanobrevibacter smithii activate human dendritic cells. PLoS One [Internet]. [cited 2025 Jan 28];9. Available from: https://pubmed.ncbi.nlm.nih.gov/24915454/ Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 19 May, 2025 Read the published version in Molecular Biology Reports → Version 1 posted Editorial decision: Revision requested 25 Apr, 2025 Reviews received at journal 31 Mar, 2025 Reviewers agreed at journal 28 Mar, 2025 Reviewers invited by journal 20 Mar, 2025 Editor assigned by journal 20 Mar, 2025 Submission checks completed at journal 20 Mar, 2025 First submitted to journal 17 Mar, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6248246","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":435941312,"identity":"832c6e26-9553-4a57-8048-b18f9f6ab554","order_by":0,"name":"Denise Mafra","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA1UlEQVRIiWNgGAWjYDCCAwwMHxgYanj4QZyEAuK0MM5gYDgmJ9kA0mJAvBZmY4MDIB4xWviuHX7YXFHDlrj5/OrEDw8MGOT5xQ7g1yJ5O82w8cwxmcRtN95ulgA6zHDm7AT8WgxuJ5g/bGBjA2o5uwGkJQEoQkhL+sfGhn/MiZtnnN38g0gtOYaNjW1A7/P3biPOFsnbOYWNjX3H5CRu8G6zSDCQIOwXvtvpGxsbvgGjsv/s5ps/Kmzk+aUJaEEACbBKCWKVgwD/AVJUj4JRMApGwUgCAN3nS9AwBGULAAAAAElFTkSuQmCC","orcid":"","institution":"Fluminense Federal University (UFF)","correspondingAuthor":true,"prefix":"","firstName":"Denise","middleName":"","lastName":"Mafra","suffix":""},{"id":435941313,"identity":"f95ed1c6-b145-4b59-a449-7ca75731877c","order_by":1,"name":"Livia Alvarenga","email":"","orcid":"","institution":"Fluminense Federal University (UFF)","correspondingAuthor":false,"prefix":"","firstName":"Livia","middleName":"","lastName":"Alvarenga","suffix":""},{"id":435941314,"identity":"d0449781-2b9d-4d93-bf56-ddacd3b3afe5","order_by":2,"name":"Ludmila Cardozo","email":"","orcid":"","institution":"Fluminense Federal University (UFF)","correspondingAuthor":false,"prefix":"","firstName":"Ludmila","middleName":"","lastName":"Cardozo","suffix":""},{"id":435941315,"identity":"b541026d-3066-4901-b66e-ddeb997367cb","order_by":3,"name":"Júnia Schultz","email":"","orcid":"","institution":"King Abdullah University of Science and Technology (KAUST)","correspondingAuthor":false,"prefix":"","firstName":"Júnia","middleName":"","lastName":"Schultz","suffix":""},{"id":435941316,"identity":"2812ede8-739e-432c-b18a-524d4e0f2e0d","order_by":4,"name":"Alexandre Soares Rosado","email":"","orcid":"","institution":"King Abdullah University of Science and Technology (KAUST)","correspondingAuthor":false,"prefix":"","firstName":"Alexandre","middleName":"Soares","lastName":"Rosado","suffix":""},{"id":435941317,"identity":"45d4f08e-b316-4813-8475-61c3159e2f4d","order_by":5,"name":"Natália A. Borges","email":"","orcid":"","institution":"State University of Rio de Janeiro (UERJ)","correspondingAuthor":false,"prefix":"","firstName":"Natália","middleName":"A.","lastName":"Borges","suffix":""}],"badges":[],"createdAt":"2025-03-18 01:08:13","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6248246/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6248246/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s11033-025-10562-8","type":"published","date":"2025-05-19T15:57:51+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":79648929,"identity":"0e5bfe06-daec-46be-a1f8-a5380ee84aa9","added_by":"auto","created_at":"2025-04-01 07:34:22","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":220079,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eCorrelogram at the phylum level with NLRP3 inflammasome and IL-1β in patients with CKD undergoing hemodialysis. \u003c/strong\u003eAbbreviations: interleukin 1 beta (IL1-β), interleukin 6 (IL6), mRNA for NOD-like receptor protein 3 (NLRP3). \u0026nbsp;Only significant correlations (p \u0026lt; 0.05) are displayed in the squares (positive = blue; negative = red).\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6248246/v1/904ffd76a2d03eef731f1b24.png"},{"id":79648595,"identity":"190bba78-7358-47e2-881c-035939e9fd27","added_by":"auto","created_at":"2025-04-01 07:26:22","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":140060,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eCorrelogram of the specific genus with NLRP3 inflammasome and IL-1β in patients with CKD undergoing hemodialysis. \u003c/strong\u003eAbbreviations: interleukin 1 beta (IL1-β), mRNA for NOD-like receptor protein 3 (NLRP3). Only significant correlations (p \u0026lt; 0.05) are displayed in the squares (positive = blue; negative = red).\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6248246/v1/e467b83259b4c3ee2bc8fcd5.png"},{"id":79648930,"identity":"439ec814-b4c2-41b4-b63b-32ce020caa07","added_by":"auto","created_at":"2025-04-01 07:34:22","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":108400,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eCorrelogram at the genus of archaea showing the NLRP3 inflammasome and IL-1β in patients with CKD undergoing hemodialysis. \u003c/strong\u003eAbbreviations: interleukin 1 beta (IL-1β), interleukin 6 (IL-6), mRNA for NOD-like receptor protein 3 (NLRP3).\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6248246/v1/0b6031c1da03e58e3b943c4c.png"},{"id":83460651,"identity":"4eb4a69c-7b31-499b-a427-654c06699e91","added_by":"auto","created_at":"2025-05-26 16:13:14","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1284185,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6248246/v1/deaac2c7-6d9c-4ee0-9c3c-db4318e8a743.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Gut Microbiota and NLRP3 Inflammasome Activation in Hemodialysis Patients: Exploring the Link with Systemic Inflammation","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThe interest in the instigating role of gut microbiota in maintaining host homeostasis and regulating the immune system has driven several studies [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. In nephrology, preclinical studies since the 1990s have demonstrated the effects of uremia on intestinal permeability, gut microbiota, and endotoxemia in patients with chronic kidney disease (CKD) [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Since then, scientific and technological advances have deepened our understanding of the relationship between gut microbiota and CKD. Metagenome-wide analyses of two cohorts of hemodialysis CKD patients revealed alterations in the gut microbiome, characterized by 348 differentially abundant species. These included increased levels of \u003cem\u003eBlautia spp\u003c/em\u003e., \u003cem\u003eDorea spp., Eggerthellaceae\u003c/em\u003e, and decreased \u003cem\u003ePrevotella\u003c/em\u003e and \u003cem\u003eRoseburia\u003c/em\u003e species [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Key toxin-contributing species were identified, and microbial signatures associated with hemodialysis CKD patients correlated with disease progression in non-dialyzed CKD patients [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eGut dysbiosis \u0026ndash; marked by perturbations in composition, diversity, and function of the gut microbiota \u0026ndash; is now recognized as a determinant factor disrupting the host\u0026rsquo;s homeostatic relationship. More recently, an association between gut dysbiosis and increased mortality in hemodialysis patients has been reported [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Evidence suggests that unbalanced gut microbiota disrupts intestinal homeostasis through interactions with the gut-associated lymphoid tissue (GALT), exposing the host microorganisms and their metabolites. This exposure triggers systemic inflammation, a key mediator linking gut dysbiosis to adverse outcomes in CKD [\u003cspan additionalcitationids=\"CR7\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eSeveral inflammatory pathways contribute to CKD pathology, including the complex cytoplasmatic proteins belonging to the innate immune system called inflammasomes. Among these, the Nod-Like Receptor Pyrin domain containing 3 (NLRP3) inflammasome plays a pivotal role in the inflammation upon receiving signals from bacteria and their metabolites, such as uremic toxins. Through pattern recognition receptors (PRRs), cells from the innate immune system recognize damage-associated molecular patterns (DAMPs) and pathogen-associated molecular patterns (PAMPs), such as lipopolysaccharide (LPS), peptidoglycan, and bacterial RNA. These signals induce NIMA-related kinase 7 (NEK7) to bind with NLRP3 and initiate inflammasome assembly [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe activation of the NLRP3 inflammasome results in the conversion of procaspase-1 into its active form, caspase-1. As an effector protein of the NLRP3 inflammasome, caspase-1 processes pro-IL-1β and pro-IL-18 into their active forms, IL-1β and IL-18, respectively, and also cleaves gasdermin D (GSDMD), into its active form, which then drives GSDMD-mediated pyroptosis and cell damage [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Dysregulated NLRP3 inflammasome activity in response to microbial components increases epithelial permeability and interleukin production, particularly IL-1β. Elevated IL-1β levels amplify systemic inflammation, stimulate downstream cytokine release, promote kidney inflammation, and drive Th17 cell differentiation and IL‑17 production. Excessive IL-1β secretion can lead to tissue damage and widespread inflammatory responses, posing significant risks to the host [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003ePreclinical studies have reinforced the hypothesis that gut microbiota interact with the NLRP3 inflammasome to activate it [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. While research has focused mainly on bacterial species contributing to inflammation, archaea also represent a stable yet underexplored component of the human gut microbiota[\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn mice, the microplastic intervention (environmental contaminants) negatively affected the gut microbiota, lowering the relative abundances of \u003cem\u003eDubosoella\u003c/em\u003e and increasing \u003cem\u003eBacteroides, Clostridia, Desulfovibrio, Enterorhabdus\u003c/em\u003e, and \u003cem\u003eGemella\u003c/em\u003e. This disruption of gut microbiota was linked to NLRP3 activation in the liver [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Similarly, Hong \u003cem\u003eet al.\u003c/em\u003e (2024) reported that treatment with \u003cem\u003eRuminococcus gnavus\u003c/em\u003e in KK-Ay mice for eight weeks disrupted gut microbiota homeostasis, altered the expression of tight junction proteins, and impaired kidney function. Additionally, this intervention led to increased expression of NLRP3, contributing to an inflammatory state [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAdditionally, studies on human proximal tubule epithelial cells have demonstrated that exposure to the uremic toxin indoxyl sulfate (IS) induces NLRP3 inflammasome activation, leading to increased expression of NLRP3, caspase-1, and IL-1β, as well as enhanced IL-1β secretion and caspase-1 activity. Furthermore, IS prompted the production of intracellular reactive oxygen species. In the same study, the authors observed a rising trend in the expression of the NLRP3 inflammasome in the CKD animal model, correlated with the plasma levels of IS, kynurenic acid, and hippuric acid [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAlthough there is increasing evidence supporting a connection between gut microbiota and NLRP3 inflammasome responses, data on CKD patients remain limited. This pilot study assesses the potential relationship between NLRP3 inflammasome activation, IL-1β expression, and gut microbiota composition in CKD patients undergoing hemodialysis.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStudy design and participants\u003c/h2\u003e \u003cp\u003eThis cross-sectional pilot study included hemodialysis patients, male and female, \u0026ge; 18 years old, with arteriovenous fistula for vascular access in the upper limb, at least 6 months on dialysis. Patients with inflammatory diseases, cancer, AIDS, autoimmune disease, smokers, pregnancy, and patients using catabolic drugs, antioxidant vitamin supplements, pre-, pro, and symbiotic, and antibiotics in the last 3 months before the start of this study were excluded. Dialysis duration was 3-4.5 hours per session, three times weekly, with a blood flow\u0026thinsp;\u0026gt;\u0026thinsp;250mL/min and a dialysate flow of 500mL/min. The Ethics Committee of Medicine Faculty of Federal Fluminense University reviewed and approved the study protocol (CAAE 47703315.6.0000.5243). All the patients who agreed to participate in the study signed an informed consent form.\u003c/p\u003e \u003cp\u003eBody mass index (BMI) was calculated by dividing dry body weight (kg) by height squared (m). The kinetic index of dialysis (Kt/V) quantifies hemodialysis treatment and was calculated using the single-pool Daugirdas formula, with a reference value of \u0026gt;\u0026thinsp;1.2 according to NKF-KDOQI (National Kidney Foundation Kidney Disease Outcomes Quality Initiatives) guidelines. A dietary 24-hour recall for 3 days (including a dialysis day, a non-dialysis day, and a weekend day) was used to compute the average dietary intake, utilizing nutritional software (Nutwin, developed by the Department of Nutrition at the Federal University of S\u0026atilde;o Paulo, UNIFESP). Demographic and clinical data were obtained by analyzing medical records and conducting interviews.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eBlood Sample Collection and Biochemical Analysis\u003c/h3\u003e\n\u003cp\u003eBlood samples were drawn before a regular hemodialysis session. Venous blood samples were drawn into an EDTA syringe (1.0 mg/mL). The blood was centrifuged (3,500 rpm for 15 minutes, at 4\u0026ordm;C) to obtain plasma, distributed in 1.5mL polypropylene Eppendorf \u0026reg; tubes, separated into aliquots, and stored at -80\u0026deg;C for further analysis. An aliquot was used for whole blood analysis to obtain peripheral blood mononuclear cells (PBMCs), according to Cardozo et al. (2016)[\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Routine biochemical analysis was performed using BioClin (Bio-clin BS-120 Chemistry Analyze).\u003c/p\u003e\n\u003ch3\u003eNLRP3 inflammasome and IL-1β analysis\u003c/h3\u003e\n\u003cp\u003eThe mRNA expression of NLRP3 and IL-1β was assessed from PBMCs using real-time quantitative polymerase chain reaction (qPCR). PBMCs were isolated from blood, and RNA was extracted using the SV Total RNA Isolation System (Promega\u0026reg;). cDNA was synthesized using the High-Capacity cDNA Reverse Transcription kit (Thermo Fisher\u0026reg;). A TaqMan Gene Expression assay (Thermo Fisher\u0026reg;) was performed to detect the expression of NLRP3 mRNA (Hs00918082_m1), IL-1β mRNA (Hs00918082_m1) and the control gene GAPDH (Hs02758991_g1). The ABI Prism 7500 Sequence Detection System (Applied Biosystems\u0026reg;) and standard cycling conditions were used for PCR amplification. The expression level was calculated using the ΔΔCT (delta cycle threshold) method.\u003c/p\u003e\n\u003ch3\u003eGut microbiota analysis\u003c/h3\u003e\n\u003cp\u003eThe patients received a sterile stool collection tube and instructions for collecting stool samples. The collected stool samples were stored at -20\u0026deg;C until further processing. According to the manufacturer's protocol, DNA extraction was performed using the Quick-DNA Fecal/Soil Microbe Miniprep Kit (Zymo Research). The concentration and quality of the total DNA were determined using a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA).\u003c/p\u003e \u003cp\u003eThe V4 region of the 16S rRNA gene was sequenced for the gut microbiota analysis. Firstly, the region was amplified via PCR using the primers 515F (5\u0026rsquo;-GTGYCAGCMGCCGCGGTAA-3\u0026rsquo;) and 806R (5\u0026rsquo;-GGACTACNVGGGTWTCTAAT-3\u0026rsquo;), with universal Illumina adapters. The thermocycling conditions included an initial denaturation at 94\u0026deg;C for 3 minutes, followed by 32 cycles of 94\u0026deg;C for 45 seconds, 50\u0026deg;C for 1 minute, and 72\u0026deg;C for 90 seconds, with a final extension at 72\u0026deg;C for 10 minutes. The resulting amplicons were barcoded, pooled, and sequenced on the Illumina NovaSeq PE250 platform (targeting 0.1\u0026nbsp;million raw reads per sample) according to the manufacturer\u0026rsquo;s protocol at Novogene (California, USA). A total of 12 samples were subjected to amplicon sequencing.\u003c/p\u003e \u003cp\u003eThe raw 16S rRNA gene sequencing reads were processed using the USEARCH pipeline (v. 11). Sequence abundances were determined, and Operational Taxonomic Units (OTUs) were clustered at 97% identity. Chimeric sequences were filtered through a denoising step to generate a Zero-radius OTU (zOTU) table. Feature and taxonomy tables, along with metadata, were exported as \u0026lsquo;phyloseq\u0026rsquo; objects for further analysis in R (v.4.1.2). For the correlation assessment, the data was then transformed using centered log ratio (CLR) transformation [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e] and submitted to Spearman\u0026rsquo;s rank correlation coefficient, with a significance threshold of p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) using the \u0026lsquo;corrplot\u0026rsquo; R package. All plots and graphs were generated with the R package \u0026lsquo;ggplot2\u0026rsquo; v.4.1.2.\u003c/p\u003e\n\u003ch3\u003eStatistical Analyses\u003c/h3\u003e\n\u003cp\u003eData were expressed as median and quartile intervals or mean and SD as appropriate. Statistical significance was accepted as p\u0026thinsp;\u0026lt;\u0026thinsp;0.05. Statistical analyses were performed using the statistical software SPSS 24.0. The Wilk test verified the distribution of descriptive variables in the present study. According to the distribution of explanatory variables, the results are expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD (standard deviation) or median (quartile intervals). Statistical analyses were performed using SPSS 24.0 (SPSS, Inc., Chicago, IL, USA).\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eThe study involved 12 CKD patients undergoing hemodialysis. Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e presents the general, biochemical, inflammatory, and dietary characteristics of HD patients. Hypertensive nephrosclerosis was the primary etiological factor of CKD. According to BMI classification, 50% of the patients were overweight, and none were malnourished. All patients showed adequate Kt/V.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe HD patients' general, biochemical, inflammatory, and dietary characteristics.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"2\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eParameters\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOverall (n\u0026thinsp;=\u0026thinsp;12)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGeneral features\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFemale/Male (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e50/50\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAge (years)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e59.2\u0026thinsp;\u0026plusmn;\u0026thinsp;13.4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eKt/V\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHD vintage (months)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e48.6 (20.1\u0026ndash;77.2)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBody mass index (kg/m\u0026sup2;)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e26.6\u0026thinsp;\u0026plusmn;\u0026thinsp;3.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTranscription factors\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003emRNA NLPR3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003emRNA Interleukin 1-beta\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.4 (0.8\u0026ndash;2.7)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRoutine Biochemical\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTotal Cholesterol (mg/dL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e143.3\u0026thinsp;\u0026plusmn;\u0026thinsp;33.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTriglycerides (mg/dL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e119 (93.6-169.1)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHigh-Density Lipoprotein (mg/dL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e42.5\u0026thinsp;\u0026plusmn;\u0026thinsp;11.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLow-Density Lipoprotein (mg/dL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e73.6\u0026thinsp;\u0026plusmn;\u0026thinsp;26.3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCalcium (mg/dL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePhosphorus (mg/dL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAlbumin (mg/dL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePotassium (mg/dL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eUrea (mg/dL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e148.7\u0026thinsp;\u0026plusmn;\u0026thinsp;34.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHemoglobin (mg/dL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eParathormone (mg/dL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e517 (381\u0026ndash;1199)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ehigh-sensitivity C-reactive protein (mg/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.5 (0.2\u0026ndash;9.2)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDietary assessments\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEnergy (Kcal/day)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1500\u0026thinsp;\u0026plusmn;\u0026thinsp;479\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCarbohydrate (g/day)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e214\u0026thinsp;\u0026plusmn;\u0026thinsp;66.8\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eProtein (g/day)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e70\u0026thinsp;\u0026plusmn;\u0026thinsp;22.8\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLipid (g/day)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e42\u0026thinsp;\u0026plusmn;\u0026thinsp;18.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePhosphorus (mg/day)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1006\u0026thinsp;\u0026plusmn;\u0026thinsp;343\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePotassium (mg/day)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1904\u0026thinsp;\u0026plusmn;\u0026thinsp;639\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"2\"\u003eNon-normally distributed variables are presented as median (25th percentile and 75th percentile) and normally distributed variables as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD. Kt/V: kinetic index of dialysis; NLPR3: Nod-Like Receptor Pyrin domain containing 3\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eRegarding the gut microbiota and its relationship with inflammation markers, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e indicates that the abundance of Fusobacteria phylum is negatively correlated (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) with the mRNA expression levels of NLRP3 and IL-1β. In contrast, members of the phylum Lentisphaerae exhibited a positive correlation with the mRNA expression levels of NLRP3 and IL-1β (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Additionally, a positive correlation was observed between IL-1β and the phylum Spirochaetes. As expected, NLRP3 and IL-1β expression levels were positively correlated (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e illustrates the correlation between gut microbial genera and inflammatory markers. The relative abundance of \u003cem\u003eFusobacterium\u003c/em\u003e was negatively correlated with the mRNA expression levels of NLRP3 and IL-1β (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Additionally, the relative abundance of \u003cem\u003eErysipelatoclostridium\u003c/em\u003e and \u003cem\u003eVictivallis\u003c/em\u003e positively correlates with the mRNA expression level of NLRP3 (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Furthermore, the relative abundances of \u003cem\u003eAbscicoccus, Colidextribacter, Desulfovibrio, Fournierella, Lawsonibacter, Ruminococcus\u003c/em\u003e, and \u003cem\u003eVictivallis\u003c/em\u003e were positively correlated with IL-1β mRNA expression (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e highlights the significant positive correlation between IL-1β expression and the archaeal genus \u003cem\u003eMethanosphaera\u003c/em\u003e in CKD patients undergoing hemodialysis.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThis study investigated the association between gut microbiota and the NLRP3 inflammasome in patients with CKD undergoing hemodialysis. Our results suggest that the Lentisphaerae and Spirochaetes phyla, as well as members of the genera \u003cem\u003eErysipelatoclostridium\u003c/em\u003e, \u003cem\u003eVictivallis\u003c/em\u003e, \u003cem\u003eAbscoccus\u003c/em\u003e, \u003cem\u003eColidextribacter\u003c/em\u003e, \u003cem\u003eDesulfovibrio\u003c/em\u003e, \u003cem\u003eFournierella\u003c/em\u003e, \u003cem\u003eLawsonibacter\u003c/em\u003e, and \u003cem\u003eRuminococcus\u003c/em\u003e may play a role in activating the NLRP3 inflammasome and IL-1β expression. Also, \u003cem\u003eMethanosphaera\u003c/em\u003e, a genus of Archaea, was positively correlated with IL-1β expression, potentially contributing to the inflammation linked to gut dysbiosis in these patients.\u003c/p\u003e \u003cp\u003eIdentifying the constituent members of the gut microbiota may help us understand the structure and function of the gut microbial community characteristic of specific conditions. In this study, we observed that the Lentisphaerae phylum was positively correlated with NLRP3 and IL-1β. Lentisphaerae is a phylum associated with ischemic stroke lesions [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e] and potentially linked to myasthenia gravis [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. One study showed that after one year of lifestyle intervention involving the Mediterranean diet, individuals with nonalcoholic fatty liver disease (NAFLD) demonstrated a decrease in the Lentisphaerae phylum, suggesting that this genus may be involved in inflammatory pathways [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. \u003cem\u003eVictivallis\u003c/em\u003e belongs to the phylum \u003cem\u003eLentisphaerae\u003c/em\u003e, which was positively associated with NLRP3 inflammasome and IL-1β expression in this study. Prior research indicated that \u003cem\u003eVictivallis\u003c/em\u003e may be related to inflammatory factors in acute respiratory distress syndrome [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. \u003cem\u003eVictivallis\u003c/em\u003e levels were significantly higher in individuals with colorectal cancer compared to healthy controls [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Furthermore, a Mendelian randomization analysis by Miao \u003cem\u003eet al.\u003c/em\u003e (2024) demonstrated that the genus \u003cem\u003eVictivallis\u003c/em\u003e had a causal relationship with hypertension[\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eOur study also suggests a potential role for Spirochaetes in the expression of IL-1β. Spirochaetes play an important role in the polymicrobial infections that lead to periodontitis. Oral spirochaetes have been shown to activate both innate and adaptive immune responses. Buford \u003cem\u003eet al.\u003c/em\u003e (2018) found that the relative abundance of Spirochaetes was significantly lower in older adults compared to healthy young individuals. However, a positive correlation exists between Spirochaetes and Insulin-like growth factor-1 (IGF-1). It is important to note that the phylum is a broad taxonomic category that includes a diverse range of microorganisms with various functions and roles [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003eAbsicoccus\u003c/em\u003e, part of the \u003cem\u003eErysipelotrichaceae\u003c/em\u003e family, was described in 2020 [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e], and its role in inflammation remains unclear. However, the \u003cem\u003eErysipelotrichaceae\u003c/em\u003e family is associated with the metabolism of L-carnitine from dietary sources, resulting in increased circulating levels of trimethylamine N-oxide (TMAO) and heightened levels of inflammatory cytokines such as IL-1 β, IL-6, tumor necrosis factor-alpha (TNF-α), and TNF-β [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Additionally, exposure to pollutants like lead has been shown to affect the abundance of pathogenic bacterial families, including Erysipelotrichaceae. This is linked to increased levels of IL-8, IL-10, IL-1, and TNF-α. These findings provide valuable insights for further investigation into the role of gender in inflammation and chronic non-communicable diseases, including CKD [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe \u003cem\u003eColidextribacter\u003c/em\u003e genus was recently isolated from the human right colon. However, little is known about its function in metabolic pathways associated with human health [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. A Crohn's and Colitis Canada Genetic Environmental Microbial (CCC-GEM) cohort showed that individuals with impaired barrier function had an increased abundance of \u003cem\u003eColidextribacter\u003c/em\u003e, among other taxonomic changes [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. In an experimental model of liver fibrosis, \u003cem\u003eColidextribacter\u003c/em\u003e was identified as a harmful bacteria associated with damage to the intestinal barrier [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eContrary to our findings, a study in rats with NAFLD found a negative correlation between \u003cem\u003eColidextribacter\u003c/em\u003e and serum TNF-α [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. Another study in rats with hyperuricemia also identified \u003cem\u003eColidextribacter\u003c/em\u003e as a beneficial bacterium that increases with marine fish-derived peptide supplementation, alongside a decrease in inflammatory factors (TNF-α, IL-6, and IL-10) [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. Finally, an experimental study involving Shen-Ling-Bai-Zhu-San, a well-known formulation in traditional Chinese medicine, showed that its use inhibits NLRP3, IL-1β, and IL-18 expression levels. This finding is linked to restoring the dysfunctional intestinal microbiota in colitis-affected mice, which includes an increase in the abundance of bacteria that produce short-chain fatty acids, such as \u003cem\u003eColidextribacter\u003c/em\u003e[\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. Contradictions persist in the literature regarding the harmful or beneficial roles of the \u003cem\u003eColidextribacter\u003c/em\u003e genus.\u003c/p\u003e \u003cp\u003eThe genus \u003cem\u003eDesulfovibrio\u003c/em\u003e consists of sulfate-reducing bacteria that inhabit the human intestine, with their numbers increasing under conditions linked to microbial dysbiosis and inflammation [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. A greater abundance of \u003cem\u003eDesulfovibrio\u003c/em\u003e correlates with increased intestinal epithelial barrier permeability [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Additionally, caspase-1, IL-1β, and IL-18 expressions also elevate \u003cem\u003eDesulfovibrio\u003c/em\u003e abundance. \u003cem\u003eDesulfovibrio\u003c/em\u003e flagellins trigger the inflammatory response and the release of cytokines and caspases through the activation of nucleotide-binding oligomerization domain-like receptor (NLR) family of apoptosis inhibitory proteins (NAIP) / NLR family caspase activation and recruitment domain-containing proteins 4 (NLRC4) inflammasome, which contributes to intestinal damage[\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. Moreover, a study in rats indicated intestinal modulation by lycopene, evidenced by a decrease in hepatic protein expressions of NLRP3, Pro-Caspase-1, Caspase-1, and factor nuclear kappa B (NF-ΚB), which is associated with lower fecal levels of \u003cem\u003eDesulfovibrio\u003c/em\u003e [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eWang \u003cem\u003eet al.\u003c/em\u003e (2022) reported that \u003cem\u003eFournierella\u003c/em\u003e contributes to worsening intestinal inflammation by disrupting bile secretion and tryptophan metabolism, which affects intestinal permeability and results in diarrhea in rabbits[\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. However, other studies present contradictory findings, as calves suffering from damp-heat diarrhea, a common condition in Chinese dairy farms, showed a decrease in the abundance of the genus \u003cem\u003eFournierella\u003c/em\u003e [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003eLawsonibacter\u003c/em\u003e is a Gram-positive, anaerobic, non-motile, non-spore-forming, and asaccharolytic bacterium. One study examined the changes in fecal microbiota following Roux-en-Y gastric bypass in obese patients. The results indicated improved patient health, reduced inflammation, and notable alterations in the intestinal microbiome, including a decrease in the \u003cem\u003eLawsonibacter\u003c/em\u003e genus, among other genera [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. In contrast, a diet rich in whole corn grains for seven weeks enhanced lambs\u0026rsquo; microbiota by increasing the Lawsonibacter genus's relative abundance in the colonic mucosa [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. Fecal microbiota transplantation was examined in irritable bowel syndrome (IBS) patients. In addition to reducing bloating and severity, bacteria such as Lawsonibacter were enriched by week 12 [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003eRuminococcus\u003c/em\u003e species are strictly anaerobic, gram-positive, and non-motile cocci that do not produce endospores, and it is essential to the microbial communities of ruminants and humans. They require fermentable carbohydrates for their growth and are a critical source of short-chain fatty acid (SCFA) production [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. Profiles of \u003cem\u003eRuminococcus bromii\u003c/em\u003e in healthy individuals showed increased abundance when consuming diets rich in resistant starch, indicating that these bacteria may be vital for the fermentation of complex carbohydrates in the large intestine [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. In rats with ulcerative colitis that were subjected to Kui jie tong (KJT), a herbal from traditional Chinese medicine, it was observed that KJT inhibited the expression of NLRP3, caspase-1, and cleaved-caspase-1 mRNA and protein in colon tissues. Furthermore, KJT reduced the expression of IL-1β, IL-18, and IL-33 levels in colon tissue and serum, and added to these findings, it also increased unclassified_g__Ruminococcus_1 levels [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. However, enrichment of Lachnospiraceae in allergic children with overgrowth of \u003cem\u003eRuminococcus gnavus\u003c/em\u003e was associated with respiratory allergies. In the same study, fed mice with purified \u003cem\u003eR. gnavus\u003c/em\u003e developed histological evidence of airway inflammation and stimulated cytokine secretion [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn a model of lipopolysaccharide-induced diarrhea in mice, \u003cem\u003eRuminococcus\u003c/em\u003e showed a positive correlation with inflammatory cytokines. It was negatively correlated with tight junction proteins and the expressions of Caspase-1 and NLRP3 [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. Oral administration of \u003cem\u003eR. gnavus\u003c/em\u003e adversely affected the kidneys in mice, leading to increased levels of nitrogen, creatinine, proteinuria, uremic toxins, and inflammatory markers (NLRP3 and IL-6) [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003eMethanosphaera\u003c/em\u003e is an archaeal genus recognized as a vital component of the human microbiome. These organisms play a key role in global carbon cycling due to their ability to produce methane, a potent greenhouse gas, as a byproduct of their energy production. It is believed that the composition of the gut methanogen community contributes to variations in methane emissions. Moreover, there is increasing evidence that methanogens and/or the methane they produce may significantly affect human health and disease [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. The molecular interactions between archaea and the immune system and their potential role in inflammation remain poorly understood [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e]. Notably, an increased abundance of \u003cem\u003eMethanosphaera spp.\u003c/em\u003e has been found in individuals with inflammatory bowel disease [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e]. Specifically, Bang \u003cem\u003eet al.\u003c/em\u003e (2014) demonstrated that the archaeal species \u003cem\u003eM. stadtmanae\u003c/em\u003e induced the release of IL-1β in monocyte-derived dendritic cells[\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. Additionally, \u003cem\u003eM. stadtmanae\u003c/em\u003e was shown to activate the NLRP3 inflammasome in human monocytic BLaER1 knockout cells. More research is necessary to understand better the inflammatory responses triggered by archaea [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eWe acknowledge the limitations of this study. Due to the cross-sectional design, a causal relationship between gut microbiota and NLRP3 inflammasome activation could not be established. Factors such as other inflammatory biomarkers, metabolites, and diet may have influenced the observed associations. Additionally, the small sample size may limit the generalizability of the findings. Future studies should include power analysis, considering the NLRP3 inflammasome components investigated here.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn conclusion, gut microbiota components in hemodialysis patients may significantly activate the NLRP3 inflammasome. The interaction between specific gut microbiota taxa and NLRP3 activation could contribute to disrupting the gut barrier and systemic inflammation. However, further investigations are needed to clarify the precise mechanisms underlying the interactions between gut microbiota and the NLRP3 inflammasome in individuals with CKD. Our findings provide valuable insights into the mechanisms driving inflammation caused by gut dysbiosis in CKD.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003e\u003cstrong\u003e16S rRNA\u003c/strong\u003e \u0026ndash; 16S ribosomal RNA\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBMI\u003c/strong\u003e \u0026ndash; Body Mass Index\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCCC-GEM\u003c/strong\u003e \u0026ndash; Crohn\u0026apos;s and Colitis Canada Genetic Environmental Microbial\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCKD\u003c/strong\u003e \u0026ndash; Chronic Kidney Disease\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCLR\u003c/strong\u003e \u0026ndash; Centered Log Ratio\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDAMPs\u003c/strong\u003e \u0026ndash; Damage-Associated Molecular Patterns\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEDTA\u003c/strong\u003e \u0026ndash; Ethylenediaminetetraacetic Acid\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGAPDH\u003c/strong\u003e \u0026ndash; Glyceraldehyde-3-Phosphate Dehydrogenase\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGALT\u003c/strong\u003e \u0026ndash; Gut-Associated Lymphoid Tissue\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGSDMD\u003c/strong\u003e \u0026ndash; Gasdermin D\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHD\u003c/strong\u003e \u0026ndash; Hemodialysis\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIGF-1\u003c/strong\u003e \u0026ndash; Insulin-like Growth Factor-1\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIL-1\u003c/strong\u003e\u003cstrong\u003e\u0026beta;\u003c/strong\u003e \u0026ndash; Interleukin-1 Beta\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIL-10\u003c/strong\u003e \u0026ndash; Interleukin-10\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIL-17\u003c/strong\u003e \u0026ndash; Interleukin-17\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIL-18\u003c/strong\u003e \u0026ndash; Interleukin-18\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIL-33\u003c/strong\u003e \u0026ndash; Interleukin-33\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIL-6\u003c/strong\u003e \u0026ndash; Interleukin-6\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIL-8\u003c/strong\u003e \u0026ndash; Interleukin-8\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIS\u003c/strong\u003e \u0026ndash; Indoxyl Sulfate\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eKt/V\u003c/strong\u003e \u0026ndash; Kinetic Index of Dialysis\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eKJT\u003c/strong\u003e \u0026ndash; Kui jie tong\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNKF-KDOQI\u003c/strong\u003e \u0026ndash; National Kidney Foundation Kidney Disease Outcomes Quality Initiatives\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNIMA\u003c/strong\u003e \u0026ndash; Never In Mitosis A\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNEK7\u003c/strong\u003e \u0026ndash; NIMA-related Kinase 7\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNLRP3\u003c/strong\u003e \u0026ndash; Nod-Like Receptor Pyrin Domain Containing 3\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eOTUs\u003c/strong\u003e \u0026ndash; Operational Taxonomic Units\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePBMCs\u003c/strong\u003e \u0026ndash; Peripheral Blood Mononuclear Cells\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePCR\u003c/strong\u003e \u0026ndash; Polymerase Chain Reaction\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePAMPs\u003c/strong\u003e \u0026ndash; Pathogen-Associated Molecular Patterns\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePRRs\u003c/strong\u003e \u0026ndash; Pattern Recognition Receptors\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eqPCR\u003c/strong\u003e \u0026ndash; Quantitative Polymerase Chain Reaction\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSD\u003c/strong\u003e \u0026ndash; Standard Deviation\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSCFA\u003c/strong\u003e \u0026ndash; Short-Chain Fatty Acids\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSPSS\u003c/strong\u003e \u0026ndash; Statistical Package for the Social Sciences\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTMAO\u003c/strong\u003e \u0026ndash; Trimethylamine N-oxide\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTNF-\u0026alpha;\u003c/strong\u003e \u0026ndash; Tumor Necrosis Factor-alpha\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTNF-\u0026beta;\u003c/strong\u003e \u0026ndash; Tumor Necrosis Factor-beta\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTh17\u003c/strong\u003e \u0026ndash; T Helper 17 Cells\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ezOTU\u003c/strong\u003e \u0026ndash; Zero-radius Operational Taxonomic Unit\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eThere are no conflicts of interest to declare.\u003c/p\u003e\n\u003cp\u003eThe study protocol was reviewed and approved by the institutional ethics committee. All patients who agreed to participate in the study signed an informed consent form.\u003c/p\u003e\n\u003cp\u003eThis work was supported by Prof. Alexandre Soares Rosado\u0026rsquo;s KAUST Baseline Grant (BAS/1/1096-01-01).\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eD.M. and N.A.B. designed the research, L.A. and L.C. conducted the study and drafted the initial manuscript, J.S and A.S.R. conducted the experiments, D.M. and N.A.B. revised the manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eHou K, Wu ZX, Chen XY, Wang JQ, Zhang D, Xiao C et al (2022) Microbiota in health and diseases. Signal Transduct Target Ther [Internet]. [cited 2025 Feb 3];7. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/35461318/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/35461318/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKrukowski H, Valkenburg S, Madella AM, Garssen J, van Bergenhenegouwen J, Overbeek SA et al (2023) Gut microbiome studies in CKD: opportunities, pitfalls and therapeutic potential. Nat Rev Nephrol [Internet]. [cited 2025 Feb 3];19:87\u0026ndash;101. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/36357577/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/36357577/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHida M, Aiba Y, Sawamura S, Suzuki N, Satoh T, Koga Y (1996) Inhibition of the accumulation of uremic toxins in the blood and their precursors in the feces after oral administration of Lebenin, a lactic acid bacteria preparation, to uremic patients undergoing hemodialysis. Nephron [Internet]. [cited 2025 Feb 3];74:349\u0026ndash;55. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/8893154/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/8893154/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVaziri ND, Wong J, Pahl M, Piceno YM, Yuan J, Desantis TZ et al (2013) Chronic kidney disease alters intestinal microbial flora. Kidney Int [Internet]. [cited 2025 Feb 3];83:308\u0026ndash;15. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/22992469/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/22992469/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhang P, Wang X, Li S, Cao X, Zou J, Fang Y et al (2023) Metagenome-wide analysis uncovers gut microbial signatures and implicates taxon-specific functions in end-stage renal disease. Genome Biol [Internet]. [cited 2025 Jan 28];24. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/37828586/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/37828586/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLin TY, Wu PH, Lin YT, Hung SC Gut dysbiosis and mortality in hemodialysis patients. NPJ Biofilms Microbiomes [Internet]. 2021 [cited 2025 Jan 28];7. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/33658514/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/33658514/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSchroder K, Tschopp J (2010) The inflammasomes. Cell [Internet]. [cited 2025 Jan 28];140:821\u0026ndash;32. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/20303873/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/20303873/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZheng X, Wan J, Tan G (2023) The mechanisms of NLRP3 inflammasome/pyroptosis activation and their role in diabetic retinopathy. Front Immunol [Internet]. [cited 2025 Jan 28];14. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/37180116/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/37180116/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu F, Gao C Regulation of the Inflammasome Activation by Ubiquitination Machinery. Adv Exp Med Biol [Internet]. 2024 [cited 2025 Jan 28];1466:123\u0026ndash;34. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/39546140/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/39546140/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAnders HJ Of Inflammasomes and Alarmins: IL-1β and IL-1α in Kidney Disease. J Am Soc Nephrol [Internet]. 2016 [cited 2025 Jan 28];27:2564\u0026ndash;75. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/27516236/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/27516236/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eXu R, Cao J, wen, Lv H li, Geng Y, Guo Myao (2024) Polyethylene microplastics induced gut microbiota dysbiosis leading to liver injury via the TLR2/NF-κB/NLRP3 pathway in mice. Sci Total Environ [Internet]. [cited 2025 Jan 28];917. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/38286276/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/38286276/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHong J, Fu T, Liu W, Du Y, Bu J, Wei G et al (2024) Specific Alternation of Gut Microbiota and the Role of Ruminococcus gnavus in the Development of Diabetic Nephropathy. J Microbiol Biotechnol [Internet]. [cited 2025 Jan 28];34:547\u0026ndash;61. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/38346799/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/38346799/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMohammadzadeh R, Mahnert A, Duller S, Moissl-Eichinger C (2022) Archaeal key-residents within the human microbiome: characteristics, interactions and involvement in health and disease. Curr Opin Microbiol [Internet]. [cited 2025 Jan 28];67. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/35427870/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/35427870/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMihajlovic M, Krebber MM, Yang Y, Ahmed S, Lozovanu V, Andreeva D et al (2021) Protein-Bound Uremic Toxins Induce Reactive Oxygen Species-Dependent and Inflammasome-Mediated IL-1β Production in Kidney Proximal Tubule Cells. Biomedicines [Internet]. [cited 2025 Jan 28];9. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/34680443/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/34680443/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCardozo LFMF, Stockler-Pinto MB, Mafra D (2016) Brazil nut consumption modulates Nrf2 expression in hemodialysis patients: A pilot study. Mol Nutr Food Res [Internet]. [cited 2025 Jan 28];60:1719\u0026ndash;24. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/26992136/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/26992136/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAitchison J The Statistical Analysis of Compositional Data. Journal of the Royal Statistical Society: Series B (Methodological) [Internet]. 1982 [cited 2025 Feb 3];44:139\u0026ndash;60. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://onlinelibrary.wiley.com/doi/full/\u003c/span\u003e\u003cspan address=\"https://onlinelibrary.wiley.com/doi/full/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.2517-6161.1982.tb01195.x\u003c/span\u003e\u003cspan address=\"10.1111/j.2517-6161.1982.tb01195.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZou X, Wang L, Xiao L, Wang S, Zhang L (2022) Gut microbes in cerebrovascular diseases: Gut flora imbalance, potential impact mechanisms and promising treatment strategies. Front Immunol [Internet]. [cited 2025 Jan 28];13. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/36389714/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/36389714/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShi J, Yi M, Xie S, Wang Z, Zhang X, Tan X et al (2024) Mendelian randomization study revealed a gut microbiota-neuromuscular junction axis in myasthenia gravis. Sci Rep [Internet]. [cited 2025 Jan 28];14. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/38291090/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/38291090/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eG\u0026oacute;mez-P\u0026eacute;rez AM, Ruiz-Lim\u0026oacute;n P, Salas-Salvad\u0026oacute; J, Vioque J, Corella D, Fit\u0026oacute; M et al (2023) Gut microbiota in nonalcoholic fatty liver disease: a PREDIMED-Plus trial sub analysis. Gut Microbes [Internet]. [cited 2025 Feb 3];15. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/37345236/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/37345236/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMa J, Zhu Z, Yishajiang Y, Alarjani KM, Hong L, Luo L Role of gut microbiota and inflammatory factors in acute respiratory distress syndrome: a Mendelian randomization analysis. Front Microbiol [Internet]. 2023 [cited 2025 Jan 28];14. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/38173678/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/38173678/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eS\u0026aacute;nchez-Alcoholado L, Ord\u0026oacute;\u0026ntilde;ez R, Otero A, Plaza-Andrade I, Laborda-Illanes A, Medina JA et al Gut Microbiota-Mediated Inflammation and Gut Permeability in Patients with Obesity and Colorectal Cancer. Int J Mol Sci [Internet]. 2020 [cited 2025 Jan 28];21:1\u0026ndash;20. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/32947866/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/32947866/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMiao C, Xu X, Huang S, Kong L, He Z, Wang Y et al (2024) The Causality between Gut Microbiota and Hypertension and Hypertension-related Complications: A Bidirectional Two-Sample Mendelian Randomization Analysis. Hellenic J Cardiol [Internet]. [cited 2025 Jan 28]; Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/38336261/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/38336261/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBuford TW, Carter CS, VanDerPol WJ, Chen D, Lefkowitz EJ, Eipers P et al Composition and richness of the serum microbiome differ by age and link to systemic inflammation. Geroscience [Internet]. 2018 [cited 2025 Jan 28];40:257\u0026ndash;68. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/29869736/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/29869736/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOren A, G\u0026ouml;ker M (2024) Notification that new names of prokaryotes, new combinations and new taxonomic opinions have appeared in volume 74, part 9 of the IJSEM. Int J Syst Evol Microbiol [Internet]. [cited 2025 Jan 28];74. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/39700045/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/39700045/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShin Y, Paek J, Kim H, Kook JK, Kim JS, Kim SH et al (2020) Absicoccus porci gen. nov., sp. nov., a member of the family Erysipelotrichaceae isolated from pig faeces. Int J Syst Evol Microbiol [Internet]. [cited 2025 Jan 28];70:732\u0026ndash;7. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/31702538/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/31702538/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhang X, Wu Q, Zhao Y, Yang X (2019) Decaisnea insignis Seed Oil Inhibits Trimethylamine- N-oxide Formation and Remodels Intestinal Microbiota to Alleviate Liver Dysfunction in l-Carnitine Feeding Mice. J Agric Food Chem [Internet]. [cited 2025 Jan 28];67:13082\u0026ndash;92. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/31671940/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/31671940/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu H, Zhang H, Yu Q, Zhang S, Tu X, Zhuang F et al (2023) Lead induced structural and functional damage and microbiota dysbiosis in the intestine of crucian carp (Carassius auratus). Front Microbiol [Internet]. [cited 2025 Jan 28];14. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/37731918/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/37731918/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRicaboni D, Mailhe M, Cadoret F, Vitton V, Fournier PE, Raoult D (2016) Colidextribacter massiliensis gen. nov., sp. nov., isolated from human right colon. New Microbes New Infect [Internet]. [cited 2025 Jan 28];17:27\u0026ndash;9. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/28275436/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/28275436/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLeibovitzh H, Lee SH, Xue M, Raygoza Garay JA, Hernandez-Rocha C, Madsen KL et al Altered Gut Microbiome Composition and Function Are Associated With Gut Barrier Dysfunction in Healthy Relatives of Patients With Crohn\u0026rsquo;s Disease. Gastroenterology [Internet]. 2022 [cited 2025 Jan 28];163:1364\u0026ndash;1376.e10. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/35850197/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/35850197/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHe X, Liang J, Li X, Wang Y, Zhang X, Chen D et al (2024) Dahuang zhechong pill ameliorates hepatic fibrosis by regulating gut microbiota and metabolites. J Ethnopharmacol [Internet]. [cited 2025 Jan 28];321. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/37967779/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/37967779/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu X, Zhang Y, Li W, Zhang B, Yin J, Liuqi S et al (2022) Fucoidan Ameliorated Dextran Sulfate Sodium-Induced Ulcerative Colitis by Modulating Gut Microbiota and Bile Acid Metabolism. J Agric Food Chem [Internet]. [cited 2025 Jan 28];70:14864\u0026ndash;76. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/36378195/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/36378195/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWu C, Hu Q, Peng X, Luo J, Zhang G (2023) Marine Fish Protein Peptide Regulating Potassium Oxonate-Induced Intestinal Dysfunction in Hyperuricemia Rats Helps Alleviate Kidney Inflammation. J Agric Food Chem [Internet]. [cited 2025 Jan 28];71:320\u0026ndash;30. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/36530149/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/36530149/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGao Q, Tian W, Yang H, Hu H, Zheng J, Yao X et al (2024) Shen-Ling-Bai-Zhu-San alleviates the imbalance of intestinal homeostasis in dextran sodium sulfate-induced colitis mice by regulating gut microbiota and inhibiting the NLRP3 inflammasome activation. J Ethnopharmacol [Internet]. [cited 2025 Jan 28];319. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/37704122/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/37704122/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh SB, Coffman CN, Carroll-Portillo A, Varga MG, Lin HC (2021) Notch Signaling Pathway Is Activated by Sulfate Reducing Bacteria. Front Cell Infect Microbiol [Internet]. [cited 2025 Jan 28];11. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/34336718/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/34336718/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh SB, Coffman CN, Varga MG, Carroll-Portillo A, Braun CA, Lin HC (2022) Intestinal Alkaline Phosphatase Prevents Sulfate Reducing Bacteria-Induced Increased Tight Junction Permeability by Inhibiting Snail Pathway. Front Cell Infect Microbiol [Internet]. [cited 2025 Jan 28];12. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/35694541/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/35694541/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAn Y, Zhai Z, Wang X, Ding Y, He L, Li L et al (2023) Targeting Desulfovibrio vulgaris flagellin-induced NAIP/NLRC4 inflammasome activation in macrophages attenuates ulcerative colitis. J Adv Res [Internet]. [cited 2025 Jan 28];52:219\u0026ndash;32. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/37586642/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/37586642/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGao X, Zhao X, Liu M, Zhao H, Sun Y Lycopene prevents non-alcoholic fatty liver disease through regulating hepatic NF-κB/NLRP3 inflammasome pathway and intestinal microbiota in mice fed with high-fat and high-fructose diet. Front Nutr [Internet]. 2023 [cited 2025 Jan 28];10. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/37032779/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/37032779/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang J, Zhao K, Kang Z, Wang M, Chen Y, Fan H et al The Multi-Omics Analysis Revealed a Metabolic Regulatory System of Cecum in Rabbit with Diarrhea. Animals (Basel) [Internet]. 2022 [cited 2025 Jan 28];12. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/35565618/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/35565618/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYan Z, Zhang K, Zhang K, Wang G, Wang L, Zhang J et al (2022) Integrated 16S rDNA Gene Sequencing and Untargeted Metabolomics Analyses to Investigate the Gut Microbial Composition and Plasma Metabolic Phenotype in Calves With Dampness-Heat Diarrhea. Front Vet Sci [Internet]. [cited 2025 Jan 28];9. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/35242833/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/35242833/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePrykhodko O, Burleigh S, Campanello M, Iresj\u0026ouml; BM, Zilling T, Ljungh \u0026Aring; et al Long-Term Changes to the Microbiome, Blood Lipid Profiles and IL-6 in Female and Male Swedish Patients in Response to Bariatric Roux-en-Y Gastric Bypass. Nutrients [Internet]. 2024 [cited 2025 Jan 28];16. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/38398821/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/38398821/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCheng C, Yin Y, Bian G (2022) Effects of whole maize high-grain diet feeding on colonic fermentation and bacterial community in weaned lambs. Front Microbiol [Internet]. [cited 2025 Jan 28];13. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/36569065/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/36569065/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYau YK, Su Q, Xu Z, Tang W, Ching JYL, Mak JWY et al (2023) Randomised clinical trial: Faecal microbiota transplantation for irritable bowel syndrome with diarrhoea. Aliment Pharmacol Ther [Internet]. [cited 2025 Jan 28];58:795\u0026ndash;804. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/37667968/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/37667968/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRainey FA, Janssen PH Phylogenetic analysis by 16S ribosomal DNA sequence comparison reveals two unrelated groups of species within the genus Ruminococcus. FEMS Microbiol Lett [Internet]. 1995 [cited 2025 Jan 28];129:69\u0026ndash;73. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/7781992/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/7781992/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAbell GCJ, Cooke CM, Bennett CN, Conlon MA, McOrist AL Phylotypes related to Ruminococcus bromii are abundant in the large bowel of humans and increase in response to a diet high in resistant starch. FEMS Microbiol Ecol [Internet]. 2008 [cited 2025 Jan 28];66:505\u0026ndash;15. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/18616586/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/18616586/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eXue S, Xue Y, Dou D, Wu H, Zhang P, Gao Y et al Kui Jie Tong Ameliorates Ulcerative Colitis by Regulating Gut Microbiota and NLRP3/Caspase-1 Classical Pyroptosis Signaling Pathway. Dis Markers [Internet]. 2022 [cited 2025 Jan 28];2022. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/35832643/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/35832643/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChua HH, Chou HC, Tung YL, Chiang BL, Liao CC, Liu HH et al Intestinal Dysbiosis Featuring Abundance of Ruminococcus gnavus Associates With Allergic Diseases in Infants. Gastroenterology [Internet]. 2018 [cited 2025 Jan 28];154:154\u0026ndash;67. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/28912020/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/28912020/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChen X, Kong Q, Zhao X, Zhao C, Hao P, Irshad I et al Sodium acetate/sodium butyrate alleviates lipopolysaccharide-induced diarrhea in mice via regulating the gut microbiota, inflammatory cytokines, antioxidant levels, and NLRP3/Caspase-1 signaling. Front Microbiol [Internet]. 2022 [cited 2025 Jan 28];13. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/36386709/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/36386709/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVolmer JG, McRae H, Morrison M (2023) The evolving role of methanogenic archaea in mammalian microbiomes. Front Microbiol [Internet]. [cited 2025 Jan 28];14. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/37727289/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/37727289/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMohammadzadeh R, Mahnert A, Duller S, Moissl-Eichinger C (2022) Archaeal key-residents within the human microbiome: characteristics, interactions and involvement in health and disease. Curr Opin Microbiol [Internet]. [cited 2025 Jan 28];67. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/35427870/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/35427870/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLecours PB, Marsolais D, Cormier Y, Berberi M, Hach\u0026eacute; C, Bourdages R et al (2014) Increased prevalence of Methanosphaera stadtmanae in inflammatory bowel diseases. PLoS One [Internet]. [cited 2025 Jan 28];9. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/24498365/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/24498365/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBang C, Weidenbach K, Gutsmann T, Heine H, Schmitz RA (2014) The intestinal archaea Methanosphaera stadtmanae and Methanobrevibacter smithii activate human dendritic cells. PLoS One [Internet]. [cited 2025 Jan 28];9. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/24915454/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/24915454/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"molecular-biology-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mole","sideBox":"Learn more about [Molecular Biology Reports](https://www.springer.com/journal/11033)","snPcode":"11033","submissionUrl":"https://submission.nature.com/new-submission/11033/3","title":"Molecular Biology Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"NLRP3 inflammasome, IL-1β, gut microbiota, dysbiosis, hemodialysis","lastPublishedDoi":"10.21203/rs.3.rs-6248246/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6248246/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eThe Nod-Like Receptor Pyrin domain-containing 3 (NLRP3) inflammasome is a critical sensor of bacterial signals and metabolites, initiating an inflammatory response. Chronic kidney disease (CKD) is often accompanied by systemic inflammation, which can be involved with gut dysbiosis. Considering this interplay, we aimed to explore the potential association between NLRP3 inflammasome expression and gut microbiota in CKD patients undergoing hemodialysis (HD).\u003c/p\u003e\u003ch2\u003eMethods and results\u003c/h2\u003e \u003cp\u003eThis research comprises a cross-sectional pilot study involving twelve HD patients [59.2\u0026thinsp;\u0026plusmn;\u0026thinsp;13.4 years, six women, BMI 26.6\u0026thinsp;\u0026plusmn;\u0026thinsp;3.5 kg/m\u003csup\u003e2\u003c/sup\u003e, 48.6 (20.1\u0026ndash;77.2) months on dialysis]. The gut microbiota was evaluated by the 16S ribosomal RNA gene. The mRNA expression of NLRP3 was assessed using real-time quantitative polymerase chain reaction (qPCR). Plasma levels of IL-1β were measured by ELISA. A positive correlation between mRNA expression of NLRP3 and Lentisphaerae phylum and with \u003cem\u003eErysipelaloclostrium\u003c/em\u003e and \u003cem\u003eVictivallis\u003c/em\u003e genus (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) was observed. The IL-1β mRNA expression was positively correlated with Lentisphaerae and Spirochaetes phylum. Also, there was a positive correlation with the relative abundance of the genera \u003cem\u003eErysipelaloclostrium, Absicoccus, Colidextribacter, Desulfovibrio, Fournierella, Lawsonibacter, Ruminococcus\u003c/em\u003e, and \u003cem\u003eVictivallis.\u003c/em\u003e Regarding Archaea, IL-1β mRNA expression was positively correlated with \u003cem\u003eMethanobrevibacter.\u003c/em\u003e\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eIn CDK patients undergoing hemodialysis, gut microbiota may be involved in NLRP3 activation and IL-1β expression, contributing to inflammation.\u003c/p\u003e","manuscriptTitle":"Gut Microbiota and NLRP3 Inflammasome Activation in Hemodialysis Patients: Exploring the Link with Systemic Inflammation","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-01 07:26:18","doi":"10.21203/rs.3.rs-6248246/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-04-25T17:23:44+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-03-31T11:59:20+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"65996660859890558728890920934128773599","date":"2025-03-28T13:20:19+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-03-20T14:29:02+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-03-20T11:47:06+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-03-20T11:45:18+00:00","index":"","fulltext":""},{"type":"submitted","content":"Molecular Biology Reports","date":"2025-03-18T01:01:53+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"molecular-biology-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mole","sideBox":"Learn more about [Molecular Biology Reports](https://www.springer.com/journal/11033)","snPcode":"11033","submissionUrl":"https://submission.nature.com/new-submission/11033/3","title":"Molecular Biology Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"eae64afa-9503-4bf5-9b48-677c980220a3","owner":[],"postedDate":"April 1st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-05-26T16:09:31+00:00","versionOfRecord":{"articleIdentity":"rs-6248246","link":"https://doi.org/10.1007/s11033-025-10562-8","journal":{"identity":"molecular-biology-reports","isVorOnly":false,"title":"Molecular Biology Reports"},"publishedOn":"2025-05-19 15:57:51","publishedOnDateReadable":"May 19th, 2025"},"versionCreatedAt":"2025-04-01 07:26:18","video":"","vorDoi":"10.1007/s11033-025-10562-8","vorDoiUrl":"https://doi.org/10.1007/s11033-025-10562-8","workflowStages":[]},"version":"v1","identity":"rs-6248246","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6248246","identity":"rs-6248246","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}