The role of nitric oxide pathway in arginine transport and growth of IPEC-1 cells

Leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5) and B-cell-specific Moloney murine leukemia virus insertion site 1 (BMI1) are markers of fast-cycling and quiescent intestinal stem cells, respectively. To determine the functions of these proteins in large animals, we investigated their effects on the proliferation of intestinal epithelial cells from pigs. Our results indicated that LGR5 and BMI1 are highly conserved proteins and that the pig proteins have greater homology with the human proteins than do mouse proteins. Overexpression of either LGR5 or BMI1 promoted cell proliferation and WNT/β-catenin signaling in pig intestinal epithelial cells (IPEC-J2). Moreover, the activation of WNT/β-catenin signaling by recombinant human WNT3A protein increased cell proliferation and LGR5 and BMI1 protein levels. Conversely, inhibition of WNT/β-catenin signaling using XAV939 reduced cell proliferation and LGR5 and BMI1 protein levels. This is the first report that LGR5 and BMI1 can increase proliferation of pig intestinal epithelial cells by activating WNT/β-catenin signaling.

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“…Cell proliferation was determined by the cell count and MTT assays, as previously described [ 41 , 42 ].…”

Section: Methodsmentioning

confidence: 99%

LGR5 and BMI1 Increase Pig Intestinal Epithelial Cell Proliferation by Stimulating WNT/β-Catenin Signaling

Wang

Chen

et al. 2018

IJMS

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“…Under conditions of l ‐NAME treatment, our results indicate that l ‐NAME can also suppress the expression of p‐Akt. Similarly, some other studies have reported that the inhibition of eNOS also affects the PI3K–Akt pathway in turn to decrease expression level .…”

Section: Discussionmentioning

confidence: 53%

Inhibition of endothelial nitric oxide synthase in cholangiocarcinoma cell lines – a new strategy for therapy

et al. 2018

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The isoform of nitric oxide synthase (NOS) found in endothelial cells (eNOS) plays a crucial role in vasodilation. We recently reported the activation of eNOS in cholangiocarcinoma (CCA) tissues and cell lines. Moreover, we also reported that the abundance of eNOS and phosphorylated eNOS (p‐eNOS), as well as its upstream regulator proteins, is significantly associated with the metastatic status of CCA patients. However, the function of eNOS in CCA progression has not been addressed. Therefore, the present study aimed to investigate the function of eNOS involved in the migration and invasion ability of CCA cell lines. The results reveal that eNOS activation significantly increases migration and invasion ability of CCA cells via the up‐regulation of phosphorylated vasodilator‐stimulated protein (p‐VASP). A combination treatment with recombinant human vascular endothelial growth factor C and eNOS inhibitor (Nω‐nitro‐l‐arginine methyl ester hydrochloride) resulted in the down‐regulation of p‐VASP, as well as a decreased migration and invasion ability of the CCA cell line. Thus, this work suggests that eNOS can serve as an attractive target to inhibit the progression of CCA.

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“…As eNOS is membrane bound, Shin et al [ 47 ] determined that endothelial cells are reliant on extracellular (i.e., plasma) bioavailability of L-arginine: circulating plasma concentration of L-arginine is approximately 50–100 µM. In contrast, RBC-NOS which is found within the intracellular and membrane compartments [ 6 ], may rapidly uptake 200 µM L-arginine, in approximately 10 min [ 48 ] which stimulates NOS phosphorylation through the PI3 kinase/Akt kinase pathway [ 49 , 50 ].…”

Section: Discussionmentioning

confidence: 99%

“…Given that RBC-NOS Serine1177 phosphorylation occurs within the reductase domain, it was not clear whether L-NAME would affect RBC-NOS Serine1177 phosphorylation per se. Nevertheless, several studies have demonstrated that L-arginine plays a role in the phosphorylation and activation of the PI3 kinase/Akt kinase pathway [ 49 , 50 ]. Given the present findings, it is plausible that the specific NOS inhibitor, L-NAME, may also indirectly inhibit NOS phosphorylation via inhibition of upstream NOS regulatory proteins (e.g., PI3/Akt kinase), at least using the concentration employed in the present study.…”

Section: Discussionmentioning

confidence: 99%

Shear Stress and RBC-NOS Serine1177 Phosphorylation in Humans: A Dose Response

Horobin

Sabapathy

Kuck

et al. 2021

Life

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Red blood cells (RBC) express a nitric oxide synthase isoform (RBC-NOS) that appears dependent on shear stress for Serine1177 phosphorylation. Whether this protein is equally activated by varied shears in the physiological range is less described. Here, we explored RBC-NOS Serine1177 phosphorylation in response to shear stress levels reflective of in vivo conditions. Whole blood samples were exposed to specific magnitudes of shear stress (0.5, 1.5, 4.5, 13.5 Pa) for discrete exposure times (1, 10, 30 min). Thereafter, RBC-NOS Serine1177 phosphorylation was measured utilising immunofluorescence labelling. Shear stress exposure at 0.5, 1.5, and 13.5 Pa significantly increased RBC-NOS Serine1177 phosphorylation following 1 min (p < 0.0001); exposure to 4.5 Pa had no effect after 1 min. RBC-NOS Serine1177 phosphorylation was significantly increased following 10 min at each magnitude of shear stress (0.5, 1.5, 13.5 Pa, p < 0.0001; 4.5 Pa, p = 0.0042). Shear stress exposure for 30 min significantly increased RBC-NOS Serine1177 phosphorylation at 0.5 Pa and 13.5 Pa (p < 0.0001). We found that RBC-NOS phosphorylation via shear stress is non-linear and differs for a given magnitude and duration of exposure. This study provides a new understanding of the discrete relation between RBC-NOS and shear stress.

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The role of nitric oxide pathway in arginine transport and growth of IPEC-1 cells

Cited by 7 publications

References 28 publications

LGR5 and BMI1 Increase Pig Intestinal Epithelial Cell Proliferation by Stimulating WNT/β-Catenin Signaling

LGR5 and BMI1 Increase Pig Intestinal Epithelial Cell Proliferation by Stimulating WNT/β-Catenin Signaling

Inhibition of endothelial nitric oxide synthase in cholangiocarcinoma cell lines – a new strategy for therapy

Shear Stress and RBC-NOS Serine1177 Phosphorylation in Humans: A Dose Response

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