Gliadin Peptide P31-43 Induces mTOR/NFkβ Activation and Reduces Autophagy: The Role of Lactobacillus paracasei CBA L74 Postbiotc

Conte, M; Nigro, Federica; Porpora, Monia; Bellomo, Claudia; Furone, Francesca; Budelli, Andrea; Nigro, Roberto; Barone, Maria Vittoria; Nanayakkara, Merlin

doi:10.3390/ijms23073655

Cited by 17 publications

(19 citation statements)

References 49 publications

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“…By far, several studies have reported the regulation of autophagy by postbiotics. For instance, the culture supernatant of Lacticaseibacillus paracasei induced autophagy in Caco‐2 cells (Conte et al, 2022); postbiotics originated from Limosilactobacillus fermentum mitigated hepatotoxicity by activating autophagy (Dinić et al, 2017). However, few study have explored their role in eliminating pathogens and the underlying mechanism remains obscure.…”

Section: Introductionmentioning

confidence: 99%

Lactobacillus plantarum postbiotics trigger AMPK‐dependent autophagy to suppress Salmonella intracellular infection and NLRP3 inflammasome activation

Wu¹,

Hu²,

Shu³

et al. 2023

Journal Cellular Physiology

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We previously found that Lactobacillus plantarum (LP)‐derived postbiotics protected animals against Salmonella infection, but the molecular mechanism remains obscure. This study clarified the mechanisms from the perspective of autophagy. Intestinal porcine epithelial cells (IPEC‐J2) were pretreated with LP‐derived postbiotics (the culture supernatant, LPC; or heat‐killed bacteria, LPB), and then challenged with Salmonella enterica Typhimurium (ST). Results showed that LP postbiotics markedly triggered autophagy under ST infection, as indicated by the increased LC3 and Beclin1 and the decreased p62 levels. Meanwhile, LP postbiotics (particularly LPC) exhibited a strong capacity of inhibiting ST adhesion, invasion and replication. Pretreatment with the autophagy inhibitor 3‐methyladenine (3‐MA) led to a significant decrease of autophagy and the aggravated infection, indicating the importance of autophagy in LP postbiotics‐mediated Salmonella elimination. LP postbiotics (especially LPB) significantly suppressed ST‐induced inflammation by modulating inflammatory cytokines (the increased interleukin (IL)‐4 and IL‐10, and decreased tumor necrosis factor‐α (TNF), IL‐1β, IL‐6 and IL‐18). Furthermore, LP postbiotics inhibited NOD‐like receptor protein 3 (NLRP3) inflammasome activation, as evidenced by the decreased levels of NLRP3, Caspase‐1 and apoptosis‐associated speck‐like protein containing a caspase recruitment domain (ASC). Deficits in autophagy resulted in an increase of inflammatory response and inflammasome activation. Finally, we found that both LPC and LPB triggered AMP‐activated protein kinase (AMPK) signaling pathway to induce autophagy, and this was further confirmed by AMPK RNA interference. The intracellular infection and NLRP3 inflammasome were aggravated after AMPK knockdown. In summary, LP postbiotics trigger AMPK‐mediated autophagy to suppress Salmonella intracellular infection and NLRP3 inflammasome in IPEC‐J2 cells. Our findings highlight the effectiveness of postbiotics, and provide a new strategy for preventing Salmonella infection.

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Section: Introductionmentioning

confidence: 99%

Lactobacillus plantarum postbiotics trigger AMPK‐dependent autophagy to suppress Salmonella intracellular infection and NLRP3 inflammasome activation

Wu¹,

Hu²,

Shu³

et al. 2023

Journal Cellular Physiology

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“…When mTOR is dephosphorylated, autophagy is induced, whereas the opposite happens when mTOR is phosphorylated ( 9 , 39 ). Therefore, in this study, the effects of gliadin peptides P31-43 and PTG on the autophagy pathway were evaluated in Caco-2 cells ( Figure 2 ).…”

Section: Resultsmentioning

confidence: 99%

“…The pH after incubation was ~7. The supernatant (metabiotic) was filtered through a 0.22-mm filter to remove the bacterial bodies ( 9 ). This preparation (metabiotic) without the bacterial bodies was used for the experiments and is referred to as “postbiotic” throughout the text.…”

Section: Methodsmentioning

confidence: 99%

The protective role of Lactobacillus rhamnosus GG postbiotic on the alteration of autophagy and inflammation pathways induced by gliadin in intestinal models

et al. 2023

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Celiac disease (CD) is an autoimmune enteropathy caused by an abnormal immune response to gliadin peptides in genetically predisposed individuals. For people with CD, the only available therapy thus far is the lifelong necessity for a gluten-free diet (GFD). Innovative therapies include probiotics and postbiotics as dietary supplements, both of which may benefit the host. Therefore, the present study aimed to investigate the possible beneficial effects of the postbiotic Lactobacillus rhamnosus GG (LGG) in preventing the effects induced by indigested gliadin peptides on the intestinal epithelium. In this study, these effects on the mTOR pathway, autophagic function, and inflammation have been evaluated. Furthermore, in this study, we stimulated the Caco-2 cells with the undigested gliadin peptide (P31-43) and with the crude gliadin peptic-tryptic peptides (PTG) and pretreated the samples with LGG postbiotics (ATCC 53103) (1 × 108). In this study, the effects induced by gliadin before and after pretreatment have also been investigated. The phosphorylation levels of mTOR, p70S6K, and p4EBP-1 were increased after treatment with PTG and P31-43, indicating that the intestinal epithelial cells responded to the gliadin peptides by activating the mTOR pathway. Moreover, in this study, an increase in the phosphorylation of NF-κβ was observed. Pretreatment with LGG postbiotic prevented both the activation of the mTOR pathway and the NF-κβ phosphorylation. In addition, P31-43 reduced LC3II staining, and the postbiotic treatment was able to prevent this reduction. Subsequently, to evaluate the inflammation in a more complex intestinal model, the intestinal organoids derived from celiac disease patient biopsies (GCD-CD) and controls (CTR) were cultured. Stimulation with peptide 31-43 in the CD intestinal organoids induced NF-κβ activation, and pretreatment with LGG postbiotic could prevent it. These data showed that the LGG postbiotic can prevent the P31-43-mediated increase in inflammation in both Caco-2 cells and in intestinal organoids derived from CD patients.

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“…Since the cytosolic Ca 2+ level is considered a signal to activate autophagy [ 31 ], we hypothesize that a reduced [Ca 2+ ]i and a low release of Ca 2+ from ER could contribute to accumulate autophagosomes in CD cells treated with THP. Recently, it has been demonstrated that in Caco-2 cells gliadin peptides were able to impair autophagy [ 32 , 33 ]. Therefore, it is possible to speculate that gliadin peptides could exacerbate the CD phenotype regarding defective autophagy.…”

Section: Discussionmentioning

confidence: 99%

Peculiar Ca2+ Homeostasis, ER Stress, Autophagy, and TG2 Modulation in Celiac Disease Patient-Derived Cells

Sposito

Secondo²,

Romanelli

et al. 2023

IJMS

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Celiac disease (CD) is an inflammatory intestinal disease caused by the ingestion of gluten-containing cereals by genetically predisposed individuals. Constitutive differences between cells from CD patients and control subjects, including levels of protein phosphorylation, alterations of vesicular trafficking, and regulation of type 2 transglutaminase (TG2), have been reported. In the present work, we investigated how skin-derived fibroblasts from CD and control subjects responded to thapsigargin, an endoplasmic reticulum ER stress inducer, in an attempt to contribute to the comprehension of molecular features of the CD cellular phenotype. We analyzed Ca2+ levels by single-cell video-imaging and TG2 activity by a microplate assay. Western blots and PCR analyses were employed to monitor TG2 levels and markers of ER stress and autophagy. We found that the cytosolic and ER Ca2+ level of CD cells was lower than in control cells. Treatments with thapsigargin differently activated TG2 in control and CD cells, as well as caused slightly different responses regarding the activation of ER stress and the expression of autophagic markers. On the whole, our findings identified further molecular features of the celiac cellular phenotype and highlighted that CD cells appeared less capable of adapting to a stress condition and responding in a physiological way.

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Gliadin Peptide P31-43 Induces mTOR/NFkβ Activation and Reduces Autophagy: The Role of Lactobacillus paracasei CBA L74 Postbiotc

Cited by 17 publications

References 49 publications

Lactobacillus plantarum postbiotics trigger AMPK‐dependent autophagy to suppress Salmonella intracellular infection and NLRP3 inflammasome activation

Lactobacillus plantarum postbiotics trigger AMPK‐dependent autophagy to suppress Salmonella intracellular infection and NLRP3 inflammasome activation

The protective role of Lactobacillus rhamnosus GG postbiotic on the alteration of autophagy and inflammation pathways induced by gliadin in intestinal models

Peculiar Ca2+ Homeostasis, ER Stress, Autophagy, and TG2 Modulation in Celiac Disease Patient-Derived Cells

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