scholarly journals Antiviral effects of ergosterol peroxide in a pig model of porcine deltacoronavirus (PDCoV) infection involves modulation of apoptosis and tight junction in the small intestine

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Cong Duan ◽  
Junchi Wang ◽  
Yi Liu ◽  
Jialu Zhang ◽  
Jianyong Si ◽  
...  
Keyword(s):  
Small Intestine ◽  
Tight Junction ◽  
In Vitro Study ◽  
Mapk Signaling ◽  
Intestinal Lesion ◽  
Ergosterol Peroxide ◽  
Junction Protein ◽  
Induced Apoptosis

AbstractPorcine deltacoronavirus (PDCoV) is a newly discovered swine enteropathogenic coronavirus with worldwide distribution. However, efficient strategies to prevent or treat the infection remain elusive. Our in vitro study revealed that ergosterol peroxide (EP) from the mushroom Cryptoporus volvatus has efficient anti-PDCoV properties. The aim of this study is to evaluate the potential of EP as a treatment for PDCoV in vivo and elucidate the possible mechanisms. Seven-day-old piglets were infected with PDCoV by oral administration in the presence or absence of EP. Piglets infected with PDCoV were most affected, whereas administration of EP reduced diarrhea incidence, alleviated intestinal lesion, and decreased viral load in feces and tissues. EP reduced PDCoV-induced apoptosis and enhanced tight junction protein expressions in the small intestine, maintaining the integrity of the intestinal barrier. EP showed immunomodulatory effect by suppressing PDCoV-induced pro-inflammatory cytokines and the activation of IκBα and NF-κB p65, and upregulating IFN-I expression. Knockdown of p38 inhibited PDCoV replication and alleviated PDCoV-induced apoptosis, implying that EP inhibited PDCoV replication and alleviated PDCoV-induced apoptosis via p38/MAPK signaling pathway. Collectively, ergosterol peroxide can protect piglets from PDCoV, revealing the potential of EP for development as a promising strategy for treating and controlling the infection of PDCoV.

scholarly journals Layilin is critical for mediating hyaluronan 35 kDa-induced intestinal epithelial tight junction protein ZO-1 in vitro and in vivo

2018 ◽  
Vol 66 ◽  
pp. 93-109 ◽  
Author(s):  
Yeojung Kim ◽  
Gail A. West ◽  
Greeshma Ray ◽  
Sean P. Kessler ◽  
Aaron C. Petrey ◽  
...  
Keyword(s):  
Tight Junction ◽  
Tight Junction Protein ◽  
Intestinal Epithelial ◽  
Junction Protein ◽  
Epithelial Tight Junction

scholarly journals In Vivo and In Vitro Study of Immunostimulation by Leuconostoc lactis-Produced Gluco-Oligosaccharides

Molecules ◽  
2019 ◽  
Vol 24 (21) ◽  
pp. 3994 ◽  
Author(s):  
Sulhee Lee ◽  
In Ho Song ◽  
Young-Seo Park
Keyword(s):  
In Vitro Study ◽  
Treated Group ◽  
Mapk Signaling ◽  
Mitogen Activated Protein Kinase ◽  
Raw264.7 Macrophages ◽  
Donor And Acceptor ◽  
Mitogen Activated Protein ◽  
Leuconostoc Lactis

Glycosyltransferase-producing Leuconostoc lactis CCK940 produces CCK- oligosaccharides, gluco-oligosaccharide molecules, using sucrose and maltose as donor and acceptor molecules, respectively. In this study, the immunostimulatory activities of CCK-oligosaccharides on RAW264.7 macrophages and BALB/c mice were evaluated. CCK-oligosaccharides induced the expression of phosphorylated-p38, extracellular-signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK) and upregulation of phagocytic activity in RAW264.7 macrophages, suggesting their involvement in mitogen-activated protein kinase (MAPK) signaling pathway and phagocytosis. When CCK-oligosaccharides were administered to mice intraperitoneally injected with cyclophosphamide (CY), spleen indices and expressions of interleukin (IL)-6, IL–10, and tumor necrosis factor-α increased, compared with those in only CY-treated group. These findings suggest that CCK-oligosaccharides can be used as an effective immunostimulating agent.

scholarly journals Damage to intestinal barrier integrity in piglets caused by porcine reproductive and respiratory syndrome virus infection

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Jin Zhao ◽  
Shuangxiu Wan ◽  
Na Sun ◽  
Panpan Sun ◽  
Yaogui Sun ◽  
...  
Keyword(s):  
Small Intestine ◽  
Tight Junction ◽  
Intestinal Barrier ◽  
Respiratory Syndrome Virus ◽  
Jejunal Mucosa ◽  
N Protein ◽  
Cell Counts ◽  
Junction Protein ◽  
Qpcr Quantification

AbstractPorcine reproductive and respiratory syndrome (PRRS) induces respiratory disease and reproductive failure accompanied by gastroenteritis-like symptoms. The mechanism of intestinal barrier injury caused by PRRSV infection in piglets has yet to be investigated. An in vivo PRRSV-induced model was established in 30-day-old piglets by the intramuscular injection of 2 mL of 104 TCID50/mL PRRSV for 15 days. Observations of PRRSV replication and histology were conducted in the lungs and intestine, and goblet cell counts, relative MUC2 mRNA expression, and tight junction protein, proinflammatory cytokine, TLR4, MyD88, IκB and p-IκB expression were measured. PRRSV replicated in the lungs and small intestine, as demonstrated by absolute RT-qPCR quantification, and the PRRSV N protein was detected in the lung interstitium and jejunal mucosa. PRRSV infection induced both lung and gut injury, markedly decreased villus height and the villus to crypt ratio in the small intestine, and obviously increased the number of goblet cells and the relative expression of MUC2 mRNA in the jejunum. PRRSV infection aggravated the morphological depletion of tight junction proteins and increased IL-1β, IL-6, IL-8 and TNF-α expression by activating the NF-κB signalling pathway in the jejunum. PRRSV infection impaired intestinal integrity by damaging physical and immune barriers in the intestine by inducing inflammation, which may be related to the regulation of the gut-lung axis. This study also provides a new hypothesis regarding the pathogenesis of PRRSV-induced diarrhoea.

116. Contribution of claudin-11 to the inter-Sertoli cell tight junction, in vitro

2005 ◽  
Vol 17 (9) ◽  
pp. 72
Author(s):  
M. J. McCabe ◽  
P. G. Stanton
Keyword(s):  
Tight Junction ◽  
Mrna Expression ◽  
Sertoli Cell ◽  
Adhesion Molecule ◽  
Sertoli Cells ◽  
Adherens Junction ◽  
Adherens Junction Protein ◽  
Junction Protein

The inter-Sertoli cell tight junction (TJ) forms the blood testis barrier (BTB) between Sertoli cells and is composed of three major transmembrane proteins: claudin-11, occludin and junctional adhesion molecule. Formation of the BTB occurs during puberty associating with an increase in circulating gonadotrophins. Claudin-11 and occludin are hormonally regulated in vitro although their importance to the function of the TJ is unknown. The aim of this study was to investigate the contribution of claudin-11 to the inter-Sertoli cell TJ in vitro by blocking gene expression using RNA interference. Two claudin-11-specific siRNA fragments were designed for this purpose. Sertoli cells in primary culture formed stable TJs within 5 days as measured by transepithelial electrical resistance (TER). The addition of siRNA for 2 days resulted in a significant (P < 0.01) 55% (mean, SD, n = 4 cultures) decrease in TER along with a major reduction in claudin-11 localisation to the TJ as assessed by immunocytochemistry. The specificity of the siRNA was shown by the presence of extensive immunostaining of occludin and of the adherens junction protein β-catenin in the same treatments. Similarly, claudin-11 mRNA expression significantly (P < 0.01) decreased by 71% (mean, SD, n = 3 cultures) in response to both claudin-11 siRNA fragments. Occludin mRNA expression was not affected. It is concluded that claudin-11 contributes at least 55% to the function of the rat Sertoli cell TJ in vitro. It is hypothesised that the remaining 45% of TJ function can be attributed to other integral proteins, such as occludin and junctional adhesion molecule. It is expected that claudin-11 and other TJ proteins play a pivotal role in the function of the BTB in vivo with potential implications in fertility and contraception.

F173 Progestin induced apoptosis of normal huma breast cells: An in vivo and in vitro study

Maturitas ◽  
1996 ◽  
Vol 27 ◽  
pp. 97 ◽  
Author(s):  
J. Desreux ◽  
A. Noël ◽  
J.L. Thomas ◽  
A.M. Bernard ◽  
J. Paris ◽  
...  
Keyword(s):  
In Vitro Study ◽  
Vitro Study ◽  
Breast Cells ◽  
Induced Apoptosis

scholarly journals Paracingulin Regulates the Activity of Rac1 and RhoA GTPases by Recruiting Tiam1 and GEF-H1 to Epithelial Junctions

2008 ◽  
Vol 19 (10) ◽  
pp. 4442-4453 ◽  
Author(s):  
Laurent Guillemot ◽  
Serge Paschoud ◽  
Lionel Jond ◽  
Andrea Foglia ◽  
Sandra Citi
Keyword(s):  
Tight Junction ◽  
Small Gtpases ◽  
Cell Junctions ◽  
Dependent Manner ◽  
Junction Protein ◽  
Junction Formation ◽  
Epithelial Junctions ◽  
Junctional Protein

Small GTPases control key cellular events, including formation of cell–cell junctions and gene expression, and are regulated by activating and inhibiting factors. Here, we characterize the junctional protein paracingulin as a novel regulator of the activity of two small GTPases, Rac1 and RhoA, through the functional interaction with their respective activators, Tiam1 and GEF-H1. In confluent epithelial monolayers, paracingulin depletion leads to increased RhoA activity and increased expression of mRNA for the tight junction protein claudin-2. During tight junction assembly by the calcium-switch, Rac1 shows two transient peaks of activity, at earlier (10–20 min) and later (3–8 h) time points. Paracingulin depletion reduces such peaks of Rac1 activation in a Tiam1-dependent manner, resulting in a delay in junction formation. Paracingulin physically interacts with GEF-H1 and Tiam1 in vivo and in vitro, and it is required for their efficient recruitment to junctions, based on immunofluorescence and biochemical experiments. Our results provide the first description of a junctional protein that interacts with GEFs for both Rac1 and RhoA, and identify a novel molecular mechanism whereby Rac1 is activated during junction formation.

PepT1-mediated fMLP transport induces intestinal inflammation in vivo

2002 ◽  
Vol 283 (6) ◽  
pp. C1795-C1800 ◽  
Author(s):  
Marion Buyse ◽  
Annick Tsocas ◽  
Francine Walker ◽  
Didier Merlin ◽  
Andre Bado
Keyword(s):  
Small Intestine ◽  
Histological Analysis ◽  
Intestinal Inflammation ◽  
In Vitro Study ◽  
Peptide Transporter ◽  
Rat Colon ◽  
Membrane Transporter ◽  
Rat Small Intestine

In the present study, the effect of H+/peptide transporter (PepT1)-mediated N-formylmethionyl-leucyl-phenylalanine (fMLP) transport on inflammation in vivo in the rat small intestine, which expresses high PepT1 levels, and in the rat colon, which does not express PepT1, were investigated using myeloperoxidase (MPO) activity and histological analysis. We found that 10 μM fMLP perfusion in the jejunum for 4 h significantly increased MPO activity and altered the architecture of jejunal villi. In contrast, 10 μM fMLP perfusion in the colon for 4 h did not induce any inflammation. In addition, we have shown that 50 mM Gly-Gly alone did not affect basal MPO activity but completely inhibited the MPO activity induced by 10 μM fMLP in the jejunum. Together, these experiments demonstrate that 1) the differential expression of PepT1 between the small intestine and the colon plays an important role in epithelial-neutrophil interactions and 2) the inhibition of fMLP uptake by jejunal epithelial cells (expressing PepT1) reduces the neutrophil ability to move across the epithelium, in agreement with our previously published in vitro study. This report constitutes the first in vivo study showing the implication of a membrane transporter (PepT1) in intestinal inflammation.

scholarly journals Tight Junction–associated MARVEL Proteins MarvelD3, Tricellulin, and Occludin Have Distinct but Overlapping Functions

2010 ◽  
Vol 21 (7) ◽  
pp. 1200-1213 ◽  
Author(s):  
David R. Raleigh ◽  
Amanda M. Marchiando ◽  
Yong Zhang ◽  
Le Shen ◽  
Hiroyuki Sasaki ◽  
...  
Keyword(s):  
Tight Junction ◽  
Protein Interactions ◽  
Electron Microscopic Examination ◽  
Electron Microscopic ◽  
Junction Protein ◽  
Tight Junction Regulation ◽  
Sirna Knockdown ◽  
Related Proteins

In vitro studies have demonstrated that occludin and tricellulin are important for tight junction barrier function, but in vivo data suggest that loss of these proteins can be overcome. The presence of a heretofore unknown, yet related, protein could explain these observations. Here, we report marvelD3, a novel tight junction protein that, like occludin and tricellulin, contains a conserved four-transmembrane MARVEL (MAL and related proteins for vesicle trafficking and membrane link) domain. Phylogenetic tree reconstruction; analysis of RNA and protein tissue distribution; immunofluorescent and electron microscopic examination of subcellular localization; characterization of intracellular trafficking, protein interactions, dynamic behavior, and siRNA knockdown effects; and description of remodeling after in vivo immune activation show that marvelD3, occludin, and tricellulin have distinct but overlapping functions at the tight junction. Although marvelD3 is able to partially compensate for occludin or tricellulin loss, it cannot fully restore function. We conclude that marvelD3, occludin, and tricellulin define the tight junction–associated MARVEL protein family. The data further suggest that these proteins are best considered as a group with both redundant and unique contributions to epithelial function and tight junction regulation.

scholarly journals Cadmium Induced Apoptosis in MG63 Cells by Increasing ROS, Activation of p38 MAPK and Inhibition of ERK 1/2 Pathways

2015 ◽  
Vol 36 (2) ◽  
pp. 642-654 ◽  
Author(s):  
Kong-He Hu ◽  
Wen-Xue Li ◽  
Min-Ying Sun ◽  
She-Bing Zhang ◽  
Cai-Xia Fan ◽  
...  
Keyword(s):  
P38 Mapk ◽  
Mapk Signaling ◽  
Osteosarcoma Cell ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Short Term Exposure ◽  
Mg63 Cells ◽  
Induced Apoptosis

Background/Aims: Cadmium (Cd) induces apoptosis in different kinds of cells, including osteoblasts, both in vivo and in vitro. However, little is known about the mechanisms by which Cd induces apoptosis. Methods: In the present study, we used the human osteosarcoma cell line MG63, which has characteristics similar to human osteoblasts, as an in vitro model to determine the cellular mechanisms by which Cd induces apoptosis. Results: We found that short-term exposure to CdCl2 induced apoptosis in MG63 cells. Furthermore, the incubation of cells with CdCl2 significantly increased the level of phosphorylated p38MAPK and significantly decreased the phosphorylation of ERK1/2 in a concentration-dependent manner. Additionally, the inhibition of the phosphorylation of p38 MAPK by SB202190 protected MG63 cells from Cd-induced apoptosis. The incubation of MG63 cells with the ERK1/2 inhibitor PD98059 significantly increased apoptosis in MG63 cells. CdCl2 also significantly increased the intracellular levels of ROS. N-acetylcysteine (NAC) significantly reduced ROS levels and reversed the effects of CdCl2 on MAPK signaling. Conclusion: Our results suggested that Cd induced apoptosis in MG63 cells by increasing ROS, activation of p38 MAPK and inhibition of ERK1/2 pathways.

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