Human spasmolytic polypeptide is a cytoprotective agent that stimulates cell migration

doi:10.1016/0016-5085(95)90014-4

Gastroenterology

Volume 108, Issue 1, January 1995, Pages 108-116

https://doi.org/10.1016/0016-5085(95)90014-4 Get rights and content

Abstract

$Background/Aims$ : Gastric epithelium is attacked by acid, pepsin, and ingested agents. When a mucosal lesion occurs, the defect is rapidly closed by cell migration. Because spasmolytic polypeptide is rapidly produced at sites of injury, we postulated that human spasmolytic polypeptide (hSP) was important in mucosal repair. Recombinant hSP was used to test this hypothesis. $Methods$ : The ulcer healing effect of various doses of hSP administered orally and subcutaneously was examined using an indomethacin (20 mg/kg) restraint rat model of gastric damage. Stability of hSP in gastrointestinal juice was determined using size-exclusion chromatography. The effect of hSP on migration of human colonic carcinoma cell lines HT29 and SW480 was determined using collagen gel invasion and wounded monolayer assays. Proliferation was assessed using [³H]thymidine incorporation and toluidine blue staining. $Results$ : Infusions of hSP at 25 and 50 μg·kg⁻¹·h⁻¹ subcutaneously decreased gastric damage by about 50% (P < 0.01) without changing acid secretion. Oral hSP was ineffective. hSP was stable in gastrointestinal juice. hSP stimulated migration of HT29 cells but did not affect proliferation and had no effect on SW480 cells. $Conclusions$ : hSP may play a key role in the early stages of mucosal repair by stimulating the initial re-epithelialization by cell migration.

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Trefoil factors 1, 2, and 3 (TFFs) are a family of small secretory molecules involved in the protection and repair of the gastrointestinal tract (GI). TFFs maintain and restore epithelial structural integrity via transducing key signaling pathways for epithelial cell migration, proliferation, and invasion. In recent years, TFFs have emerged as key players in the pathogenesis of multiple diseases, especially cancer. Initially recognized as tumor suppressors, emerging evidence demonstrates their key role in tumor progression and metastasis, extending their actions beyond protection. However, to date, a comprehensive understanding of TFFs’ mechanism of action in tumor initiation, progression and metastasis remains obscure. The present review discusses the structural, functional and mechanistic implications of all three TFF family members in tumor progression and metastasis. Also, we have garnered information from studies on their structure and expression status in different organs, along with lessons from their specific knockout in mouse models. In addition, we highlight the emerging potential of using TFFs as a biomarker to stratify tumors for better therapeutic intervention.
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The mammalian intestinal epithelium serves as a critical barrier between the host organism and the contents of the gut, providing for regulated uptake of food and fluid as well as protection from pathogens and toxins. This single-cell lining is constantly subjected to insult and injury and must be efficiently repaired. The initial and critical phase of repair is restitution, in which epithelial cells at the wound perimeter migrate into the injured region and reseal the barrier. Failure or dysregulation of these processes is implicated in the pathogenesis of a number of intestinal and extraintestinal disease states. A thorough understanding of the regulatory mechanisms promoting and limiting the wound healing response in the gut epithelium will open new therapeutic avenues. This chapter will discuss the basic mechanisms of cell migration involved in restitution, the laboratory methods used to investigate this process, and current knowledge of the molecular and cellular mechanisms by which the organism regulates wound healing in the small intestine and colon.
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Trefoil factor 3 (TFF3) is a protease-resistant molecule secreted by goblet cells that play a role in mucosal healing by epithelial restitution, mucosal protection, enhancement of the structural integrity of the mucosal barrier, and increasing mucus viscosity [6]. It is also involved in intestinal homeostasis through potentiating epithelial restitution [7,8]. TFF1 and TFF3-overexpressing mice show a protective effect against intestinal damage and ulceration [9,10], whereas mice with deleted TFF1 or TFF3 genes show higher susceptibility to gastrointestinal injury [11,12].
Strengthening of intestinal tight junctions provides an effective barrier from the external environment. Goblet cell-derived trefoil factor 3 (TFF3) increases transepithelial resistance by upregulating the expression of tight junction proteins. Oxyresveratrol (OXY) is a hydroxyl-substituted stilbene found in the roots, leaves, stems, and fruit of many plants and known to have various biological activities. In this study, we investigated the strengthening effect of OXY on intestinal tight junctions through stimulation of TFF production in goblet cells.
We prepared conditioned medium from LS 174T goblet cells treated with OXY (GCO-CM) and investigated the effect of GCO-CM on strengthening tight junctions of Caco-2 cells. The mRNA and protein expression levels of major tight junction components (claudin-1, occludin, and ZO-1) were measured by quantitative real-time PCR and western blotting, respectively. Transepithelial electric resistance (TEER) was measured using an ohm/V meter. Monolayer permeability was evaluated by paracellular transport of fluorescein isothiocyanate-dextran.
OXY showed a strong antioxidant activity. It significantly increased the expression level of TFF3 in LS 174T goblet cells. GCO-CM prepared by treatment with 2.5, 5, and 10 μg/ml OXY did not show cytotoxicity in Caco-2 cells. GCO-CM increased the mRNA and protein expression levels of claudin-1, occludin, and ZO-1. It also significantly increased tight junction integrity and reduced permeability in a dose-dependent manner.
OXY stimulates the expression of TFF3 in goblet cells, which might increase the integrity of the intestinal tight junction barrier.
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The epithelial surfaces of the mammalian intestine interface directly with the external environment and thus continuously encounter pathogenic bacteria, fungi, viruses, and parasites. The intestinal epithelium is also closely associated with complex communities of symbiotic microorganisms. Intestinal epithelial cells are thus faced with the unique challenge of directly interacting with enormous numbers of microbes that include both pathogens and symbionts. As a result, gut epithelia have evolved an array of strategies that contribute to host immunity. This chapter considers the various mechanisms used by epithelial cells to limit microbial invasion of host tissues, shape the composition of indigenous microbial communities, and coordinate the adaptive immune response to microorganisms. Study of intestinal epithelial cells has contributed fundamental insights into intestinal immune homeostasis and has revealed how impaired epithelial cell function can contribute to inflammatory disease.
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The mucin produced in deep gastric glands appears to express the majority, if not all of the organ-specific carbohydrates, which have been shown to exert antibiotic effects on H. pylori (20). The co-expressed lectin, TFF2, has been claimed to control epithelial repair (33), but also to regulate mononuclear cell inflammatory responses in H. pylori infection (34). In support of its potential role in H. pylori defense mechanisms, an accelerated progression of gastritis to dysplasia in the pyloric antrum was reported in TFF2−/− mice infected with H. pylori (35).
Helicobacter pylori infection is the major cause of gastric cancer and remains an important health care challenge. The trefoil factor peptides are a family of small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract. H. pylori colocalizes with MUC5AC at the gastric surface epithelium, but not with MUC6 secreted in concert with TFF2 by deep gastric glands. Both components of the gastric gland secretome associate non-covalently and show increased expression upon H. pylori infection. Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. We show here that the gastric glycoform of TFF2 is a calcium-independent lectin, which binds with high specificity to O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin. The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, ¹H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. Neoglycolipids derived from the C3/C6-linked branches of the two isomers revealed highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1). Supposedly, lectin TFF2 is involved in protection of gastric epithelia via a functional relationship to defense against H. pylori launched by antibiotic α1,4-GlcNAc-capped mucin glycans. Lectin-carbohydrate interaction may have also an impact on more general functional aspects of TFF members by mediating their binding to cell signaling receptors.
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^☆: Supported by the Medical Research Council (to R.J.P. and T.M.) and the Imperial Cancer Research Fund (to A.M.H.).

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Human spasmolytic polypeptide is a cytoprotective agent that stimulates cell migration☆

Abstract

Gastroenterology

Gastroenterology

FEBS Lett

Lancet

Febs Lett

Gastroenterology

Gastroenterology

Gastric mucosal defence and repair

Epidermal growth factor (EGF/URO) induces expression of regulatory peptides in damaged human gastrointestinal tissues

J Pathol

Spasmolytic polypeptide, intestinal trefoil factor, EGF and TGFα are expressed sequentially during healing of cryoprobe-induced gastric ulcers in rats

Gut

Dissecting tumor cell invasion: epithelial cells acquire invasive properties after the loss of uvomorulin-mediated cell-cell adhesion

J Cell Biol

Trefoil peptide gene expression in small intestinal Crohns disease and dietary adaption

J Clin Gastroenterol