• Electroacupuncture
• inflammation remission
• intestinal glial cells
• ulcerative colitis
• visceral hypersensitivity

• Caco-2 cells
• ERK
• HIV-1 transgenic rat
• NFκB
• PPARγ

• Animals
• Rats
• Matrix Metalloproteinase 9/metabolism
• HIV Infections/metabolism
• HIV Infections/pathology
• Colon/drug effects
• Colon/metabolism
• Colon/pathology
• Methamphetamine/pharmacology
• Methamphetamine/toxicity
• Male
• Rats, Transgenic
• Signal Transduction/drug effects
• Signal Transduction/physiology
• Occludin/metabolism
• Humans
• Disease Models, Animal
• Intestinal Mucosa/drug effects
• Intestinal Mucosa/metabolism
• Intestinal Mucosa/pathology
• HIV-1
• Rats, Sprague-Dawley

• P30A1082151 Chicago Developmental Center for AIDS Research
• R25MH080661 NIMH NIH HHS

• enteric glia
• enteric glia communications
• enteric glia diversity
• enteric nervous system
• gastrointestinal diseases
• homeostasis
• immune cells

• Humans
• Neuroglia/physiology
• Enteric Nervous System/pathology
• Animals
• Gastrointestinal Diseases/pathology

• enteric nervous system
• gut–brain axis
• inflammatory bowel disease

• Crohn’s Disease (CD)
• electrical neuromodulation
• inflammatory bowel disease (IBD)
• ulcerative colitis (UC)

• Humans
• Inflammatory Bowel Diseases/therapy
• Inflammatory Bowel Diseases/physiopathology
• Electric Stimulation Therapy/methods
• Animals
• Vagus Nerve Stimulation/methods
• Tibial Nerve/physiology
• Quality of Life

• 2023YFC2507300 National Key R&D Program of China
• 82370543 Natural Science Foundation of China
• 2021CFB450 Natural Science Foundation of Hubei Province
• 2022020801010462 Wuhan Knowledge Innovation Special Basic Research Project
• 2022xhyn009 Science Research Foundation of Union Hospital
• 202120 Teaching Reform Project of the First Clinical College

• Enteric nervous system
• Inflammation
• Neuroimmune signaling
• Parkinson’s disease

• Humans
• Parkinson Disease/pathology
• Connectome
• Enteric Nervous System/pathology
• alpha-Synuclein
• Neurons/pathology
• Neuroglia/pathology

• R01 DK103723 NIDDK NIH HHS
• R01 DK120862 NIDDK NIH HHS

• Humans
• Crohn Disease/metabolism
• Neuroglia/pathology
• Neurons/pathology
• Enteric Nervous System/metabolism
• Enteric Nervous System/pathology

• 2023-E-13 2022 Baoshan Medical and Health Project
• BSZK-2023-Z06 The Most Important Discipline, Baoshan Medical Key Discipline and Characteristic Brand Construction Project
• RJPY-LX-004 Cultivation Funding for Clinical Scientific Research Innovation, Renji Hospital, School of Medicine, Shanghai Jiaotong University

• J774A1 M1 macrophages
• enteric glial cells
• phytocannabinoids

• Animals
• Lipopolysaccharides/pharmacology
• Lipopolysaccharides/metabolism
• Inflammation/drug therapy
• Inflammation/metabolism
• Neuroglia/metabolism
• Macrophages/metabolism
• Anti-Inflammatory Agents/pharmacology
• Anti-Inflammatory Agents/metabolism
• Mammals

• 12-05-0035 The Israeli Ministry of Agriculture grant

• colon
• immunohistochemistry
• microvessel
• mouse
• vasculature
• wheat germ agglutinin

• I01 RX001685 RRD VA
• OT2 OD024899 NIH HHS

• Brilliant blue G
• Enteric glial cell
• Ileum
• Myenteric plexus
• Probenecid

• Crohn’s disease
• auricular vagus nerve
• inflammatory bowel disease
• neuroprostheses
• transcutaneous auricular vagus nerve stimulation
• vagus nerve
• vagus nerve stimulation

• Humans
• Crohn Disease/therapy
• Vagus Nerve Stimulation/methods
• Inflammatory Bowel Diseases/therapy
• Colitis, Ulcerative
• Inflammation

• diarrhea
• enteric glia
• macrophage
• retinoic acid signaling
• vitamin A

• dysmotility
• enteric nervous system
• enteric neuroplasticity
• high-fat diet

• Clostridioides difficile infection
• cytokines
• inflammatory bowel diseases
• interferon gamma
• tumour necrosis factor alpha

• Astrocytes
• Brain-gut axis
• Cerebral I/R
• Colitis
• H(2)S
• RhoA/ROCK pathway

• Anxiety
• Depression
• Inflammatory bowel disease
• Medication
• Meta-analysis
• Neuropsychological assessment

• Axon-glia contacts
• Enteric glia
• Mitochondria
• Myenteric ganglia

• Animals
• Enteric Nervous System/cytology
• Enteric Nervous System/physiology
• Gastrointestinal Diseases/drug therapy
• Gastrointestinal Diseases/physiopathology
• Gastrointestinal Motility/physiology
• Homeostasis
• Humans
• Neuroglia/cytology
• Neuroglia/physiology
• Signal Transduction

• R01 DK103723 NIDDK NIH HHS
• R01 DK120862 NIDDK NIH HHS

During acute intestinal ischemia reperfusion (IR) injury, the intestinal epithelial barrier (IEB) function is often disrupted. Enteric glial cells (EGCs) play an important role in maintaining the integrity of IEB functions. However, how EGCs regulate IEB function under IR stimulation is unknown. The present study reveals that the adenosine A_2A receptor (A_2AR) is important for mediating the barrier-modulating roles of EGCs. A_2AR knockout (KO) experiments revealed more serious intestinal injury in A_2AR KO mice than in WT mice after IR stimulation. Moreover, A_2AR expression was significantly increased in WT mice when challenged by IR. To further investigate the role of A_2AR in IEB, we established an in vitro EGC-Caco-2 co-culture system. Hypoxia stimulation was used to mimic the process of in vivo IR. Treating EGCs with the CGS21680 A_2AR agonist attenuated hypoxia-induced intestinal epithelium damage through up-regulating ZO-1 and occludin expression in cocultured Caco-2 monolayers. Furthermore, we showed that A_2AR and metabotropic glutamate receptor 5 (mGluR5) combine to activate the PKCα-dependent pathway in conditions of hypoxia. This study shows, for the first time, that hypoxia induces A_2AR-mGluR5 interaction in EGCs to protect IEB function via the PKCα pathway.

• A2AR
• enteric glial cells
• hypoxia
• intestinal epithelial barrier
• mGluR5

Over recent years, several investigations have suggested that Parkinson’s disease (PD) can be regarded as the consequence of a bowel disorder. Indeed, gastrointestinal symptoms can occur at all stages of this neurodegenerative disease and in up to a third of cases, their onset can precede the involvement of the central nervous system. Recent data suggest that enteric glial cells (EGCs) may play a major role in PD-related gastrointestinal disturbances, as well as in the development and progression of the central disease. In addition to their trophic and structural functions, EGCs are crucial for the homeostatic control of a wide range of gastrointestinal activities. The main purpose of this review was to provide a detailed overview of the role of EGCs in intestinal PD-associated alterations, with particular regard for their participation in digestive and central inflammation as well as the dynamic interactions between glial cells and intestinal epithelial barrier. Accumulating evidence suggests that several pathological intestinal conditions, associated with an impairment of barrier permeability, may trigger dysfunctions of EGCs and their shift towards a proinflammatory phenotype. The reactive gliosis is likely responsible for PD-related neuroinflammation and the associated pathological changes in the ENS. Thus, ameliorating the efficiency of mucosal barrier, as well as avoiding IEB disruption and the related reactive gliosis, might theoretically prevent the onset of PD or, at least, counteract its progression.

• Parkinson’s disease
• enteric glial cells
• enteric immune system
• gut–brain axis
• intestinal epithelial barrier
• α-synuclein

• Enteric glial cell
• Glial-derived neurotrophic factor (GDNF)
• Inflammatory bowel disease (IBD)
• Mast cell

The role played by adenosine A_2B receptors (A_2BRs) in the regulation of enteric glial cell (EGC) functions remains unclear. This study was aimed at investigating the involvement of A_2BRs in the control of EGC functions in a model of obesity. C57BL/6 mice were fed with standard diet (SD) or high fat diet (HFD) for eight weeks. Colonic tachykininergic contractions were recorded in the presence of BAY60-6583 (A_2BRs agonist), MRS1754 (A_2BRs antagonist), and the gliotoxin fluorocitrate. Immunofluorescence distribution of HuC/D, S100β, and A_2BRs was assessed in whole mount preparations of colonic myenteric plexus. To mimic HFD, EGCs were incubated in vitro with palmitate (PA) and lipopolysaccharide (LPS), in the absence or in the presence of A_2BR ligands. Toll-like receptor 4 (TLR4) expression was assessed by Western blot analysis. Interleukin-1β (IL-1β), substance P (SP), and glial cell derived neurotrophic factor (GDNF) release were determined by enzyme-linked immunosorbent assay (ELISA) assays. MRS1754 enhanced electrically evoked tachykininergic contractions of colonic preparations from HFD mice. BAY60-6583 decreased the evoked tachykininergic contractions, with higher efficacy in HFD mice. Such effects were blunted upon incubation with fluorocitrate. In in vitro experiments on EGCs, PA and LPS increased TLR4 expression as well as IL-1β, GDNF, and SP release. Incubation with BAY60-6583 reduced TLR4 expression as well as IL-1β, GDNF, and SP release. Such effects were blunted by MRS1754. The present results suggest that A_2BRs, expressed on EGCs, participate in the modulation of enteric inflammation and altered tachykininergic responses associated with obesity, thus representing a potential therapeutic target.

• adenosine A2B receptors
• colonic motor dysfunction
• enteric glia
• enteric inflammation
• glial cell derived neurotrophic factor
• interleukin-1β
• obesity
• substance P
• tachykininergic contraction
• toll-like receptor 4

• Acetamides/pharmacology
• Aminopyridines/pharmacology
• Animals
• Body Weight/drug effects
• Cells, Cultured
• Citrates/pharmacology
• Diet, High-Fat
• Enteric Nervous System/pathology
• Feeding Behavior/drug effects
• Inflammation/pathology
• Interleukin-1beta/metabolism
• Lipopolysaccharides/pharmacology
• Male
• Mice, Inbred C57BL
• Mice, Obese
• Models, Biological
• Nerve Growth Factors/metabolism
• Neuroglia/drug effects
• Neuroglia/metabolism
• Obesity/pathology
• Palmitic Acid/pharmacology
• Purines/pharmacology
• Receptor, Adenosine A2B/metabolism
• S100 Proteins/metabolism
• Substance P/metabolism
• Tachykinins/metabolism
• Toll-Like Receptor 4/metabolism

• R01 DK113790 NIDDK NIH HHS
• R01 GM066189 NIGMS NIH HHS

• APCs, antigen-presenting cells
• BDNF, brain-derived neurotrophic factor
• BMDCs, bone marrow-derived dendritic cells
• Berberine
• CGRP, calcitonin gene-related peptide
• DSS, dextran sulfate sodium
• EGCs, enteric glial cells
• ENS, enteric nervous system
• Enteric glial cells
• Enteric nervous system
• GDNF, glial cell derived neurotrophic factor
• GFAP, glial fibrillary acidic protein
• IBD, inflammatory bowel diseases
• IECs, intestinal epithelial cells
• LMPC, lamina propria mononuclear cells
• MAPK, mitogen-activated protein kinases
• MLNs, mesenteric lymph nodes
• MPO, myeloperoxidase
• Mucosal inflammation
• UC, ulcerative colitis
• Ulcerative colitis
• VIP, vasoactive intestinal polypeptide

• Inflammation
• Ischemic stroke
• NLRP10
• NLRP12/ASC/Caspase-1
• TLR-4/NF-κB

Human and mouse studies have shown that rotavirus infection is associated with low inflammation and unaffected intestinal barrier at the time of diarrhea, properties different from most bacterial and inflammatory diseases of the gut. We showed by in vitro, ex vivo, and in vivo experiments that neurotrophic factors and 5-HT have barrier protective properties during rotavirus insult. These observations advance our understanding of how the gut barrier is protected against rotavirus and suggest that rotavirus affects the gut barrier differently from bacteria. This is the first report to show that neurotrophic factors contribute to maintain the gut epithelial barrier during viral insult.

Increased intestinal permeability has been proposed as a mechanism of rotavirus-induced diarrhea. Studies with humans and mice have, however, shown that rotavirus leaves intestinal permeability unaffected or even reduced during diarrhea, in contrast to most bacterial infections. Gastrointestinal permeability is regulated by the vagus nerve and the enteric nervous system, which is composed of neurons and enteric glial cells (EGCs). We investigated whether the vagus nerve, serotonin (5-HT), EGCs, and neurotropic factors contribute to maintaining gut barrier homeostasis during rotavirus infection. Using subdiaphragmatic vagotomized and 5-HT₃ receptor knockout mice, we found that the unaffected epithelial barrier during rotavirus infection is independent of the vagus nerve but dependent on 5-HT signaling through enteric intrinsic 5-HT₃ receptors. Immunofluorescence analysis showed that rotavirus-infected enterocytes were in close contact with EGCs and enteric neurons and that the glial cell-derived neurotrophic factor (GDNF) was strongly upregulated in enterocytes of infected mice. Moreover, rotavirus and 5-HT activated EGCs (P < 0.001). Using Ussing chambers, we found that GDNF and S-nitrosoglutathione (GSNO) led to denser epithelial barriers in small intestinal resections from noninfected mice (P < 0.01) and humans (P < 0.001) and that permeability was unaffected in rotavirus-infected mice. GSNO made the epithelial barrier denser in Caco-2 cells by increasing the expression of the tight junction protein zona occludens 1 (P < 0.001), resulting in reduced passage of fluorescein isothiocyanate dextran (P < 0.05) in rotavirus-infected monolayers. This is the first report to show that neurotropic factors contribute to maintaining the gut epithelial barrier during viral insult.

• diarrhea
• neurotrophic factors
• permeability
• rotavirus

• RAGE
• S100B
• SsnB
• dysbiosis
• nitric oxide
• p47 phox
• peroxynitrite
• sodium butyrate

• I01 CX001923 CSRD VA
• R01 MH094755 NIMH NIH HHS
• R01 ES019313 NIEHS NIH HHS
• P20 GM103641 NIGMS NIH HHS
• P01 AT003961 NCCIH NIH HHS
• R01 AT006888 NCCIH NIH HHS
• R01 AI123947 NIAID NIH HHS
• R01 AI129788 NIAID NIH HHS
• I01 BX001357 BLRD VA

• Enteric glial cells
• GFAP
• Ischemia and reperfusion
• P2X7 receptor
• S100β

• enteric glial cells
• high content analysis
• phycotoxins
• toxicity

Small intestinal Paneth cells, enteric glial cells (EGC), and goblet cells maintain gut mucosal integrity, homeostasis, and influence host physiology locally and through the gut-brain axis. Little is known about their roles during pregnancy, or how maternal malnutrition impacts these cells and their development. Pregnant mice were fed a control diet (CON), undernourished by 30% vs. control (UN), or fed a high fat diet (HF). At day 18.5 (term = 19), gut integrity and function were assessed by immunohistochemistry and qPCR. UN mothers displayed reduced mRNA expression of Paneth cell antimicrobial peptides (AMP; Lyz2, Reg3g) and an accumulation of villi goblet cells, while HF had reduced Reg3g and mucin (Muc2) mRNA and increased lysozyme protein. UN fetuses had increased mRNA expression of gut transcription factor Sox9, associated with reduced expression of maturation markers (Cdx2, Muc2), and increased expression of tight junctions (TJ; Cldn-7). HF fetuses had increased mRNA expression of EGC markers (S100b, Bfabp, Plp1), AMP (Lyz1, Defa1, Reg3g), and TJ (Cldn-3, Cldn-7), and reduced expression of an AMP-activator (Tlr4). Maternal malnutrition altered expression of genes that maintain maternal gut homeostasis, and altered fetal gut permeability, function, and development. This may have long-term implications for host-microbe interactions, immunity, and offspring gut-brain axis function.

• development
• gut barrier
• malnutrition
• pregnancy

• Animal Nutritional Physiological Phenomena
• Animals
• Disease Models, Animal
• Female
• Fetal Development
• Gastrointestinal Absorption/genetics
• Gastrointestinal Tract/metabolism
• Gastrointestinal Tract/physiopathology
• Gene Expression Regulation, Developmental
• Goblet Cells/metabolism
• Male
• Malnutrition/genetics
• Malnutrition/metabolism
• Malnutrition/physiopathology
• Maternal Nutritional Physiological Phenomena
• Mice, Inbred C57BL
• Nutritional Status
• Organogenesis
• Paneth Cells/metabolism
• Sex Factors

• acute infection
• enteric glial cells
• enteric nervous system
• enteric neurons
• toxoplasmosis

• Abdominal Pain
• Brain-Gut Axis
• Chronic Pain
• ENS
• Enteric Glia
• Glial Cells
• Inflammation

The research and the selection of novel probiotic strains from novel niches are receiving increased attention on their proclaimed health benefits to both humans and animals. This study aimed to evaluate the functional properties of Weissella strains from arid land living-hosts and to select strains with cholesterol-lowering property in vitro and in vivo, for use as probiotics. They were assessed for acid and bile tolerance, antibiotic susceptibility, membrane properties, antibacterial activity, antiadhesive effect against pathogens to host cell lines, and cholesterol assimilation in vitro. Our results showed that the majority of strains revealed resistance to gastrointestinal conditions. All the strains were nonhemolytic and sensitive to most of the tested antibiotics. They also exhibited high rates of autoaggregation and some of them showed high coaggregation with selected pathogens and high adhesion ability to two different cell lines (Caco-2 and MIM/PPk). Particularly, W. halotolerans F99, from camel feces, presented a broad antibacterial spectrum against pathogens, reduced Enterococcus faecalis and Escherichia coli adhesion to Caco-2 cells, and was found to reduce, in vitro, the cholesterol level by 49 %. Moreover, W. halotolerans F99 was evaluated for the carbohydrate utilization as well as the serum lipid metabolism effect in Wistar rats fed a high-cholesterol diet. W. halotolerans F99 showed an interesting growth on different plant-derivative oligosaccharides as sole carbon sources. Compared with rats fed a high-fat (HF) diet without Weissella administration, total serum cholesterol, low-density lipoprotein cholesterol, and triglycerides levels were significantly (p<0.001) reduced in W. halotolerans F99-treated HF rats, with no significant change in high-density lipoprotein cholesterol HDL-C levels. On the basis of these results, this is the first study to report that W. halotolerans F99, from camel feces, can be developed as cholesterol-reducing probiotic strain. Further studies may reveal their potential and possible biotechnological and probiotic applications.

• diabetes
• enteric glial cells
• enteric neurons
• neurotrophins

• enteric glial cells
• inflammation
• inflammatory bowel disease

• Cardiolipin
• Cytochrome c
• Enteric glial cells
• Lipotoxicity
• Mitochondrial ROS
• Palmitate

• Animals
• Cell Differentiation/physiology
• Cell Line
• Chemokine CXCL2/metabolism
• Coculture Techniques/methods
• Enteric Nervous System/metabolism
• Enteroendocrine Cells
• Epithelial Cells/metabolism
• Interleukin-10/metabolism
• Intestinal Mucosa/metabolism
• Intestinal Mucosa/physiology
• Intestine, Small
• Intestines/physiology
• Mice
• Mice, Inbred C57BL
• Myofibroblasts/metabolism
• Neuroglia/metabolism
• Neurons/metabolism
• Stem Cells
• Transforming Growth Factor beta1/metabolism

• R01 EB021908 NIBIB NIH HHS

• Crohn's disease
• enteric glial cells
• postoperative recurrence
• substance P
• vasoactive intestinal peptide

• Apolipoprotein E
• Enteric glial cells
• Gastroparesis
• Neural stem cells

Uncoupling between ATP overflow and extracellular adenosine formation changes purinergic signaling in post-inflammatory ileitis. Adenosine neuromodulation deficits were ascribed to feed-forward inhibition of ecto-5′-nucleotidase/CD73 by high extracellular adenine nucleotides in the inflamed ileum. Here, we hypothesized that inflammation-induced changes in cellular density may also account to unbalance the release of purines and their influence on [³H]acetylcholine release from longitudinal muscle-myenteric plexus preparations of the ileum of 2,4,6-trinitrobenzenesulfonic acid (TNBS)-treated rats. The population of S100β-positive glial cells increase, whereas Ano-1-positive interstitial cells of Cajal (ICCs) diminished, in the ileum 7-days after the inflammatory insult. In the absence of changes in the density of VAChT-positive cholinergic nerves detected by immunofluorescence confocal microscopy, the inflamed myenteric plexus released smaller amounts of [³H]acetylcholine which also became less sensitive to neuronal blockade by tetrodotoxin (1 μM). Instead, [³H]acetylcholine release was attenuated by sodium fluoroacetate (5 mM), carbenoxolone (10 μM) and A438079 (3 μM), which prevent activation of glial cells, pannexin-1 hemichannels and P2X7 receptors, respectively. Sodium fluoroacetate also decreased ATP overflow without significantly affecting the extracellular adenosine levels, thus indicating that surplus ATP release parallels reactive gliosis in post-inflammatory ileitis. Conversely, loss of ICCs may explain the lower amounts of adenosine detected in TNBS-treated preparations, since blockade of Ca_v3 (T-type) channels existing in ICCs with mibefradil (3 μM) or inhibition of the equilibrative nucleoside transporter 1 with dipyridamole (0.5 μM), both decreased extracellular adenosine. Data indicate that post-inflammatory ileitis operates a shift on purinergic neuromodulation reflecting the upregulation of ATP-releasing enteric glial cells and the depletion of ICCs accounting for decreased adenosine overflow via equilibrative nucleoside transporters.

• ATP release
• acetylcholine release
• adenosine release
• enteric glia
• interstitial cells of Cajal
• myenteric plexus
• post-inflammatory ileitis

To evaluate the measurement of zonulin level and antibodies of zonulin and other tight junction proteins in the blood of controls and celiac disease patients.

This study was conducted to assess the variability or stability of zonulin levels vs IgA and IgG antibodies against zonulin in blood samples from 18 controls at 0, 6, 24 and 30 h after blood draw. We also measured zonulin level as well as zonulin, occludin, vinculin, aquaporin 4 and glial fibrillary acidic protein antibodies in the sera of 30 patients with celiac disease and 30 controls using enzyme-linked immunosorbent assay methodology.

The serum zonulin level in 6 out of 18 subjects was low or < 2.8 ng/mL and was very close to the detection limit of the assay. The other 12 subjects had zonulin levels of > 2.8 ng/mL and showed significant fluctuation from sample to sample. Comparatively, zonulin antibody measured in all samples was highly stable and reproducible from sample to sample. Celiac disease patients showed zonulin levels with a mean of 8.5 ng/mL compared to 3.7 ng/mL in controls (P < 0.0001). Elevation of zonulin level at 2SD above the mean was demonstrated in 37% of celiac disease patients, while antibodies against zonulin, occludin and other tight junction proteins was detected in up to 86% of patients with celiac disease.

Due to its fluctuation, a single measurement of zonulin level is not recommended for assessment of intestinal barrier integrity. Measurement of IgG and IgA antibodies against zonulin, occludin, and other tight junction proteins is proposed for the evaluation of the loss of intestinal barrier integrity.

• Zonulin
• Occludin
• Tight junction protein
• Intestinal barrier integrity
• Celiac disease

• Adult
• Aquaporin 4/immunology
• Aquaporin 4/metabolism
• Biomarkers/blood
• Biomarkers/metabolism
• Celiac Disease/blood
• Celiac Disease/immunology
• Enzyme-Linked Immunosorbent Assay
• Glial Fibrillary Acidic Protein/immunology
• Glial Fibrillary Acidic Protein/metabolism
• Haptoglobins/analysis
• Haptoglobins/immunology
• Haptoglobins/metabolism
• Humans
• Immunoglobulin A/blood
• Immunoglobulin A/immunology
• Immunoglobulin A/metabolism
• Immunoglobulin G/blood
• Immunoglobulin G/immunology
• Immunoglobulin G/metabolism
• Intestinal Mucosa/metabolism
• Middle Aged
• Occludin/immunology
• Occludin/metabolism
• Permeability
• Tight Junctions/metabolism
• Time Factors
• Vinculin/immunology
• Vinculin/metabolism
• Young Adult

• Enteric glia
• Enteric nervous system
• GDNF
• Intestinal barrier integrity
• Morphine
• Opioids

Enteric glial cells (EGCs) are components of the intestinal epithelial barrier essential for regulating the enteric nervous system. Clostridium difficile is the most common cause of antibiotic-associated colitis, toxin B (TcdB) being the major virulence factor, due to its ability to breach the intestinal epithelial barrier and to act on other cell types. Here we investigated TcdB effects on EGCs and the activated molecular mechanisms. Already at 2 hours, TcdB triggered ROS formation originating from NADPH-oxidase, as demonstrated by their reduction in the presence of the NADPH-oxidase inhibitor ML171. Although EGCs mitochondria support almost completely the cellular ATP need, TcdB exerted weak effects on EGCs in terms of ATP and mitochondrial functionality, mitochondrial ROS production occurring as a late event. ROS activated the JNK signalling and overexpression of the proapoptotic Bim not followed by cytochrome c or AIF release to activate the downstream apoptotic cascade. EGCs underwent DNA fragmentation through activation of the ROS/JNK/caspase-3 axis, evidenced by the ability of ML171, N-acetylcysteine, and the JNK inhibitor SP600125 to inhibit caspase-3 or to contrast apoptosis. Therefore, TcdB aggressiveness towards EGCs is mainly restricted to the cytosolic compartment, which represents a peculiar feature, since TcdB primarily influences mitochondria in other cellular types.

• Enteric glia cell
• GFAP
• Inflammation
• LPS
• S100B