Abstract:Background
Metabolic reprogramming is a critical event for cell fate and function, making it an attractive target for clinical therapy. The function of metabolic reprogramming in Helicobacter pylori (H. pylori)-infected gastric intestinal metaplasia remained to be identified.
Methods
Xanthurenic acid (XA) was measured in gastric cancer cells treated with H. pylori or H. pylori virulence factor, respectively, and qPCR and WB were performed to detect… Show more
“…As described in the previous studies [ 24 , 25 ], after dewaxing and dehydration, the slide of intestinal organoid was subjected from antigen retrieval and blocked with goat serum for 30 min. The slips were incubated with the primary antibody overnight at 4°C, followed by incubation with Alexa-488- or Alexa 594-conjugated secondary antibodies for 1 hr at room temperature.…”
Section: Methodsmentioning
confidence: 99%
“…As described in our work [ 20 , 25 ], for IP, cells were lysed in ice-cold buffer composed of 50 mmol/L Tris–HCl (pH 7.4), 150 mmol/L NaCl, 0.1% NP-40 and protease inhibitors, followed by incubation overnight with anti-PKA, and further incubation was performed with protein A/G beads for 1 hr at 4°C. Beads were washed five times with low-salt lysis buffer, and immunoprecipitates were eluted for sodium dodecyl sulfate–polyacrylamide gel electrophoresis.…”
Section: Methodsmentioning
confidence: 99%
“…As described in our work [ 25 ], the reporter plasmid containing RANKL promoter was transfected into HT-29 cells with internal control pGL4.74 for 24 hr, and mannitol and sorbitol were used to treat for another 24 hr, the relative luciferase unit was measured according to dual-luciferase reporter assay system (Promega).…”
Objective. Microfold cells (M cells) are specific intestinal epithelial cells for monitoring and transcytosis of antigens, microorganisms, and pathogens in the intestine. However, the mechanism for M-cell development remained elusive. Materials and Methods. Real-time polymerase chain reaction, immunofluorescence, and western blotting were performed to analyze the effect of sorbitol-regulated M-cell differentiation in vivo and in vitro, and luciferase and chromatin Immunoprecipitation were used to reveal the mechanism through which sorbitol-modulated M-cell differentiation. Results. Herein, in comparison to the mannitol group (control group), we found that intestinal M-cell development was inhibited in response to sorbitol treatment as evidenced by impaired enteroids accompanying with decreased early differentiation marker Annexin 5, Marcksl1, Spib, sox8, and mature M-cell marker glycoprotein 2 expression, which was attributed to downregulation of receptor activator of nuclear factor kappa-В ligand (RANKL) expression in vivo and in vitro. Mechanically, in the M-cell model, sorbitol stimulation caused a significant upregulation of phosphodiesterase 4 (PDE4) phosphorylation, leading to decreased protein kinase A (PKA)/cAMP-response element binding protein (CREB) activation, which further resulted in CREB retention in cytosolic and attenuated CREB binds to RANKL promoter to inhibit RANKL expression. Interestingly, endogenous PKA interacted with CREB, and this interaction was destroyed by sorbitol stimulation. Most importantly, inhibition of PDE4 by dipyridamole could rescue the inhibitory effect of sorbitol on intestinal enteroids and M-cell differentiation and mature in vivo and in vitro. Conclusion. These findings suggested that sorbitol suppressed intestinal enteroids and M-cell differentiation and matured through PDE4-mediated RANKL expression; targeting to inhibit PDE4 was sufficient to induce M-cell development.
“…As described in the previous studies [ 24 , 25 ], after dewaxing and dehydration, the slide of intestinal organoid was subjected from antigen retrieval and blocked with goat serum for 30 min. The slips were incubated with the primary antibody overnight at 4°C, followed by incubation with Alexa-488- or Alexa 594-conjugated secondary antibodies for 1 hr at room temperature.…”
Section: Methodsmentioning
confidence: 99%
“…As described in our work [ 20 , 25 ], for IP, cells were lysed in ice-cold buffer composed of 50 mmol/L Tris–HCl (pH 7.4), 150 mmol/L NaCl, 0.1% NP-40 and protease inhibitors, followed by incubation overnight with anti-PKA, and further incubation was performed with protein A/G beads for 1 hr at 4°C. Beads were washed five times with low-salt lysis buffer, and immunoprecipitates were eluted for sodium dodecyl sulfate–polyacrylamide gel electrophoresis.…”
Section: Methodsmentioning
confidence: 99%
“…As described in our work [ 25 ], the reporter plasmid containing RANKL promoter was transfected into HT-29 cells with internal control pGL4.74 for 24 hr, and mannitol and sorbitol were used to treat for another 24 hr, the relative luciferase unit was measured according to dual-luciferase reporter assay system (Promega).…”
Objective. Microfold cells (M cells) are specific intestinal epithelial cells for monitoring and transcytosis of antigens, microorganisms, and pathogens in the intestine. However, the mechanism for M-cell development remained elusive. Materials and Methods. Real-time polymerase chain reaction, immunofluorescence, and western blotting were performed to analyze the effect of sorbitol-regulated M-cell differentiation in vivo and in vitro, and luciferase and chromatin Immunoprecipitation were used to reveal the mechanism through which sorbitol-modulated M-cell differentiation. Results. Herein, in comparison to the mannitol group (control group), we found that intestinal M-cell development was inhibited in response to sorbitol treatment as evidenced by impaired enteroids accompanying with decreased early differentiation marker Annexin 5, Marcksl1, Spib, sox8, and mature M-cell marker glycoprotein 2 expression, which was attributed to downregulation of receptor activator of nuclear factor kappa-В ligand (RANKL) expression in vivo and in vitro. Mechanically, in the M-cell model, sorbitol stimulation caused a significant upregulation of phosphodiesterase 4 (PDE4) phosphorylation, leading to decreased protein kinase A (PKA)/cAMP-response element binding protein (CREB) activation, which further resulted in CREB retention in cytosolic and attenuated CREB binds to RANKL promoter to inhibit RANKL expression. Interestingly, endogenous PKA interacted with CREB, and this interaction was destroyed by sorbitol stimulation. Most importantly, inhibition of PDE4 by dipyridamole could rescue the inhibitory effect of sorbitol on intestinal enteroids and M-cell differentiation and mature in vivo and in vitro. Conclusion. These findings suggested that sorbitol suppressed intestinal enteroids and M-cell differentiation and matured through PDE4-mediated RANKL expression; targeting to inhibit PDE4 was sufficient to induce M-cell development.
“…The exposure of XA inside gastric epithelial cells further upregulates caudal‐related homeobox transcription factor‐2 (CDX2) expression, leading to the activation of cyclic guanylate adenylate synthase (cGAS)‐interferon regulatory factor 3 (IRF3) nuclear localization to the KAT2 promoter. This positive feedback loop drives H. pylori ‐induced gastric intestinal metaplasia 77 …”
The microbiota actively and extensively participates in the regulation of human metabolism, playing a crucial role in the development of metabolic diseases. Helicobacter pylori (H. pylori), when colonizing gastric epithelial cells, not only induces local tissue inflammation or malignant transformation but also leads to systemic and partial changes in host metabolism. These shifts can be mediated through direct contact, toxic components, or indirect immune responses. Consequently, they influence various molecular metabolic events that impact nutritional status and iron absorption in the host. Unraveling the intricate and diverse molecular interaction links between H. pylori and human metabolism modulation is essential for understanding pathogenesis mechanisms and developing targeted treatments for related diseases. However, significant challenges persist in comprehensively understanding the complex association networks among H. pylori itself, the infected host's status, the host microbiome, and the immune response. Previous metabolomics research has indicated that H. pylori infection and eradication may selectively shape the metabolite and microbial profiles of gastric lesions. Yet, it remains largely unknown how these diverse metabolic pathways, including isovaleric acid, cholesterol, fatty acids, and phospholipids, specifically modulate gastric carcinogenesis or affect the host's serum metabolism, consequently leading to the development of metabolic‐associated diseases. The direct contribution of H. pylori to metabolisms still lacks conclusive evidence. In this review, we summarize recent advances in clinical evidence highlighting associations between chronic H. pylori infection and metabolic diseases, as well as its potential molecular regulatory patterns.
“…Glutathione peroxidase 2 (GPx2) contributes to the progression and metastasis of gastric cancer by enhancing the KYNU-Kyn-AhR signaling pathway [ 48 ]. Helicobacter pylori facilitates gastric intestinal metaplasia by activating the kynurenine pathway mediated by KAT2 through the cyclic GMP-AMP synthase (cGAS)-Interferon regulatory factor 3 (IRF3) signaling [ 49 ]. Fig.…”
Tryptophan (Trp) metabolism plays a crucial role in influencing the development of digestive system tumors. Dysregulation of Trp and its metabolites has been identified in various digestive system cancers, including esophageal, gastric, liver, colorectal, and pancreatic cancers. Aberrantly expressed Trp metabolites are associated with diverse clinical features in digestive system tumors. Moreover, the levels of these metabolites can serve as prognostic indicators and predictors of recurrence risk in patients with digestive system tumors. Trp metabolites exert their influence on tumor growth and metastasis through multiple mechanisms, including immune evasion, angiogenesis promotion, and drug resistance enhancement. Suppressing the expression of key enzymes in Trp metabolism can reduce the accumulation of these metabolites, effectively impacting their role in the promotion of tumor progression and metastasis. Strategies targeting Trp metabolism through specific enzyme inhibitors or tailored drugs exhibit considerable promise in enhancing therapeutic outcomes for digestive system tumors. In addition, integrating these approaches with immunotherapy holds the potential to further enhance treatment efficacy.
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