Macrophage COX2 mediates efferocytosis, resolution reprogramming, and intestinal epithelial repair

Subepithelial myofibroblasts are involved in the initiation and coordination of intestinal epithelial repair, but the molecular signaling pathways are largely unknown. The cellular adaptations that occur during repair range from dedifferentiation and migration to proliferation and redifferentiation, in a way that is strongly reminiscent of normal crypt-to-villus epithelial maturation. We therefore hypothesized that Wnt/β-catenin signaling may have a pivotal role in intestinal epithelial wound repair. We used the established scratch wound method in Caco-2 cells and in nontransformed NCM460 cells to monitor the effects of IL-1β-stimulated colonic myofibroblasts (CCD-18co) on intestinal epithelial repair, with immunoblotting and immunodepletion to examine the conditioned media. Conditioned media from IL-1β-stimulated, but not -untreated, myofibroblasts increased Caco-2 wound closure twofold over 24 h. IL-1β-stimulated myofibroblasts downregulated the differentiation marker sucrase-isomaltase in the Caco-2 cells, whereas the proliferation marker c-myc was upregulated. Array expression profiling identified Wnt-5a as the Wnt-related gene that was most upregulated (28-fold) by IL-1β stimulation of CCDs. Recombinant Wnt-5a enhanced proliferation of Caco-2 and NCM460 cells. In scratch assays, it increased migration of the leading edge in both cell lines. Wnt-5a immunodepletion of the IL-1β-CCD conditioned media abrogated the ability to enhance the repair. Wnt-5a often acts through a noncanonical signal transduction pathway. Further experiments supported this pathway in epithelial wound healing: IL-1β-CCD-mediated repair was not affected by the addition of the canonical Wnt antagonist Dickkopf-1. Furthermore, media from stimulated myofibroblasts (but not Wnt-5a-depleted media) increased c-jun in Caco-2 cell nuclear extracts. Myofibroblast-mediated noncanonical Wnt-5a signaling is therefore important in the dedifferentiation and migration stages of epithelial wound repair.

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The Promotive Effect of the Active Ingredients of Atractylodes macrocephala on Intestinal Epithelial Repair Through Activating Ca2+ Pathway

Natural Product Communications ◽

10.1177/1934578x211040357 ◽

2021 ◽

Vol 16 (11) ◽

pp. 1934578X2110403

Author(s):

Yan Ren ◽

Wenwen Jiang ◽

Chunli Luo ◽

Xiaohan Zhang ◽

Mingjin Huang

Keyword(s):

Active Ingredient ◽

Quantitative Polymerase Chain Reaction ◽

Enzyme Linked Immunosorbent Assay ◽

Intestinal Epithelial ◽

Active Ingredients ◽

Epithelial Repair ◽

Epithelial Restitution ◽

Anti Inflammatory ◽

Atractylodes Macrocephala ◽

Atractylenolide Iii

Atractylodes macrocephala ( AM) is a famous traditional Chinese medicine for intestinal epithelial restitution through activating Ca2+ channels. However, the roles of specific AM compositions in intestinal epithelial restitution are sparse. Therefore, this study aimed to compare the concrete effects of the 4 active ingredients (atractylon, β-eudesmol, atractylenolide II, atractylenolide III) of AM and their combination on intestinal epithelial repair and the Ca2+ pathway in intestinal epithelial cell (IEC-6) cells. First, the best combination of the 4 ingredients with an optimal mixing ratio of atractylon: β-eudesmol: atractylenolide II: atractylenolide III = 1:2:2:2 was demonstrated by a 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide orthogonal experiment. Subsequently, enzyme-linked immunosorbent assay was used to measure anti-inflammatory cytokine levels, the migratory ability was evaluated by cell scratch experiments, cell cycle analysis and [Ca2+]cyt concentration in cells were detected by flow cytometry, and the expression of the Ca2+ pathway-related genes was detected by immunofluorescence staining, quantitative polymerase chain reaction and whole blood assays. Our result showed that atractylon, β-Eudesmol, atractylenolide II, and atractylenolide III showed different abilities to promote the IEC-6 cells proliferation, migration, and the expression of anti-inflammatory cytokines interleukin (IL)-2, IL-10, and ornithine decarboxylase, as well as the intracellular [Ca2+]cyt concentration through stromal interaction molecule 1 transposition to activate Ca2+ pathway. Thereinto, atractylenolide III was the main active ingredient of AM for pro-proliferation and anti-inflammation, and the combination of 4 AM ingredients performed better beneficial effects on IEC-6 cells. Therefore, our study suggested that atractylenolide III was the active ingredient of AM for intestinal epithelial repair through activating the Ca2+ pathway, and the 4 ingredients of AM have a synergy in intestinal epithelial repair.

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Role of mTORC1 in intestinal epithelial repair and tumorigenesis

Cellular and Molecular Life Sciences ◽

10.1007/s00018-019-03085-6 ◽

2019 ◽

Vol 76 (13) ◽

pp. 2525-2546 ◽

Cited By ~ 5

Author(s):

Harleen Kaur ◽

Régis Moreau

Keyword(s):

Intestinal Epithelial ◽

Epithelial Repair

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Non-redundant functions of FAK and Pyk2 in intestinal epithelial repair

Scientific Reports ◽

10.1038/s41598-019-41116-1 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 2

Author(s):

Keena S. Thomas ◽

Katherine A. Owen ◽

Kathryn Conger ◽

Ryan A. Llewellyn ◽

Amy H. Bouton ◽

...

Keyword(s):

Intestinal Epithelial ◽

Epithelial Repair ◽

Redundant Functions

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Faculty Opinions recommendation of Peroxisome elevation induces stem cell differentiation and intestinal epithelial repair.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.737666442.793586914 ◽

2021 ◽

Author(s):

Yiorgos Apidianakis ◽

Savvas Teloni

Keyword(s):

Stem Cell ◽

Cell Differentiation ◽

Stem Cell Differentiation ◽

Intestinal Epithelial ◽

Epithelial Repair

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Activation of Wnt3a signaling stimulates intestinal epithelial repair by promoting c-Myc-regulated gene expression

AJP Cell Physiology ◽

10.1152/ajpcell.00341.2011 ◽

2012 ◽

Vol 302 (1) ◽

pp. C277-C285 ◽

Cited By ~ 20

Author(s):

Lan Liu ◽

Jaladanki N. Rao ◽

Tongtong Zou ◽

Lan Xiao ◽

Alexis Smith ◽

...

Keyword(s):

Gene Expression ◽

Epithelial Cells ◽

Intestinal Epithelial Cells ◽

Cyclin E ◽

Cell Renewal ◽

Intestinal Epithelial ◽

Epithelial Repair ◽

Myc Gene ◽

Factor C ◽

Regulated Gene Expression

In response to mucosal injury, epithelial cells modify the patterns of expressed genes to repair damaged tissue rapidly. Our previous studies have demonstrated that the transcription factor c-Myc is necessary for stimulation of epithelial cell renewal during mucosal healing, but the up-stream signaling initiating c-Myc gene expression after injury remains unknown. Wnts are cysteine-rich glycoproteins that act as short-range ligands to locally activate receptor-mediated signaling pathways and correlate with the increased expression of the c-Myc gene. The current study tested the hypothesis that Wnt3a signaling is implicated in intestinal epithelial repair after wounding by stimulating c-Myc expression. Elevated Wnt3a signaling in intestinal epithelial cells (IEC-6 line) by coculturing with stable Wnt3a-transfected fibroblasts or ectopic overexpression of the Wnt3a gene enhanced intestinal epithelial repair after wounding. This stimulatory effect on epithelial repair was prevented by silencing the Wnt coreceptor LRP6 or by c-Myc silencing. Activation of the Wnt3a signaling pathway increased β-catenin nuclear translocation by decreasing its phosphorylation and stimulated c-Myc expression during epithelial repair after wounding. In stable Wnt3a-transfected IEC-6 cells, increased levels of c-Myc were associated with an increase in expression of c-Myc-regulated genes cyclcin D1 and cyclin E, whereas c-Myc silencing inhibited expression of cyclin D1 and cyclin E and delayed epithelial repair. These results indicate that elevated Wnt3a signaling in intestinal epithelial cells after wounding stimulates epithelial repair by promoting c-Myc-regulated gene expression.

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Villus contraction aids repair of intestinal epithelium after injury

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.1989.257.2.g274 ◽

1989 ◽

Vol 257 (2) ◽

pp. G274-G283 ◽

Cited By ~ 17

Author(s):

R. Moore ◽

S. Carlson ◽

J. L. Madara

Keyword(s):

In Vitro Model ◽

Epithelial Barrier ◽

Intestinal Epithelial ◽

Villus Height ◽

Epithelial Barrier Function ◽

Epithelial Repair ◽

Vitro Model ◽

Energy Dependent ◽

Cytoplasmic Processes

A highly reproducible in vitro model of intestinal epithelial injury in guinea pig ileum was used to study the structural and functional events that accompany rapid epithelial repair. This model is characterized by denudation of the villus tip followed by rapid restitution of the epithelial barrier. Using standard electrophysiological and quantitative morphometric techniques, we found that immediately after injury the number of cells lost exceeded the number of empty cell positions on the denuded basement membrane by 100%. Concurrently, cytoplasmic processes of subepithelial myofibroblasts contained condensations of microfilaments that were not apparent in controls. Additionally, villus height was diminished immediately after injury, and progressively decreased during the restitution period. In tissues that were depleted of ATP using the uncoupler dinitrophenol and in tissues functionally denervated by tetrodotoxin, villus shortening after injury was significantly reduced. Denervation also retarded functionally and structurally defined reestablishment of epithelial barrier function. These data suggest that intestinal epithelial repair is aided by energy-dependent, neurally mediated villus contraction. We speculate that the subepithelial network of myofibroblasts is responsible for this process, which effectively minimizes the denuded surface area to be reepithelialized.

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