Enterocyte barriers

Figure 9.11 Gl membrane transport. Transport through the enterocyte barrier can be divided into active, passive and specialized transport and into the paracellular and transcellular routes. Efflux mechanisms can reduce absorption by these routes.

In culture, the human colon carcinoma cell hne Caco-2 spontaneously differentiates at confluency into polarized cells with enterocyte-like characteristics. The principle of this approach consists of following the passage of the compound of interest from the apical or lumen-like sides to the basolateral or lymph-hke sides of Caco-2 cells, thus following the absorption of the compound per se. One obhgate step for fat-soluble nutrients such as carotenoids to cross the intestinal barrier is their incorporation into CMs assembled in the enterocytes. Under normal cell culture conditions, Caco-2 cells are unable to form CMs. When supplemented with taurocholate and oleic acid, Caco-2 cells were reported to assemble and secrete CMs. ... 

J Wacher, L Salphati, LZ Benet. Active secretion and enterocyte drug metabolism barriers to drug absorption. Adv Drug Del Rev 20 99-112, 1996. 

Mucosal brush border membrane vesicles and basolateral membrane vesicles can be isolated to study solute uptake across specific enterocyte boundaries. These more isolated vesicle systems allow for investigation of solute transport across a particular membrane barrier and permit separation of membrane trans-... 

The barriers confronting oral delivery of a drug molecule are summarized in Fig. 13.1. During the absorption process, the drug must cross the enterocyte cell apical... 

From the above, it is clear that the gut wall represents more than just a physical barrier to oral drug absorption. In addition to the requirement to permeate the membrane of the enterocyte, the drug must avoid metabolism by the enzymes present in the gut wall cell as well as counter-absorptive efflux by transport proteins in the gut wall cell membrane. Metabolic enzymes expressed by the enterocyte include the cytochrome P450, glucuronyltransferases, sulfotransferases and esterases. The levels of expression of these enzymes in the small intestine can approach that of the liver. The most well-studied efflux transporter expressed by the enterocyte is P-gp. 

In order for allelochemicals to enter the body of a herbivore, absorption must occur across the gut lining. Curtailing the initial absorption of dietary allelochemicals may be a herbivore s first line of defense against plant toxins. Studies have citied the lack of absorption or metabolism of lipophilic plant secondary metabolites (i.e., terpenes), conducive to phase I or II detoxification, in the gut of terrestrial herbivores rather these compounds are excreted unchanged in the feces (Marsh et al. 2006b). While physical barriers or surfactants have been used to explain this limited adsorption in both marine and terrestrial herbivores (Lehane 1997 Barbehenn and Martin 1998 Barbehenn 2001 for review of marine herbivores, see Targett and Arnold 2001), active efflux of plant allelochemicals out of enterocytes into the gut lumen has received limited attention until now. 

When a drug has dissolved in the GI fluids and is present in solution at the site of absorption it has to pass a biological barrier, that is, the enterocytes lining the gut wall, in order to be absorbed into the body. The absorptive flux (J) can be described as a function of the permeability of the intestinal mucosa to the drug (Peff), the surface area available for absorption (SA), and the concentration gradient (AC) across the mucosa (e.g., [6], Eq. 2)... 

Cell monolayers grown on permeable culture inserts form confluent mono-layers with barrier properties and can be used for drug absorption experiments. The most well-known cell line for the in vitro determination of intestinal drug permeability is the human colon adenocarcinoma Caco-2 [20, 21], The utility of the Caco-2 cell line is due to its spontaneous differentiation to enterocytes under conventional cell culture conditions upon reaching confluency on a porous membrane to resemble the intestinal epithelium. This cell model displays small intestinal carriers, brush borders, villous cell model, tight junctions, and high resistance [22], Caco-2 cells express active transport systems, brush border enzymes, and phase I and II enzymes [22-24], Permeability models... 

Caco-2 cells cultured on filters achieve a monolayer density, and exhibit morphological characteristics similar to enterocytes such as tight intercellular junctions and highly developed microvilli.13-15,42 Transport of drugs across Caco-2 monolayers is limited by the action of biochemical and physical barriers (Figure 9.2). The biochemical component comprises drug-metabolizing enzymes, uptake transporters, and efflux transporters.17,19-23,28,30 The... 

Fuel for enterocytes and for stem cells in the crypts of villi. Maintenance of gut-associated lymphoid tissue (GALT). Maintenance of gut barrier, especially that in the colon. 

As mentioned above, the villi of the small intestine (Figure 1.2) house a dynamic, self-renewing population of the epithelial cells that includes absorptive cells (enterocytes), secretory cells, and endocrine cells. The thin lining (height 25 p,M height of the microvilli is 1.5 pM) of the columnar enterocytes is the only barrier between the intestinal lumen and the muscularis mucosa, which represents, in this context, the entire body interior. The entire epithelial lining of the intestine replaces itself every 3-5 d [128], It is the enterocyte and its neighboring cells where absorption processes occur and it will therefore be the focus of the mechanistic discussions below. 

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