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Despite the promising prospects of nanoparticles in oral drug delivery, the process of oral administration involves a complex transportation pathway that includes cellular uptake, intracellular trafficking, and exocytosis by intestinal epithelial cells, which are necessary steps for nanoparticles to enter the bloodstream and exert therapeutic effects. Current researchers have identified several crucial factors that regulate the interaction between nanoparticles and intestinal epithelial cells, including surface properties such as ligand modification, surface charge, hydrophilicity/hydrophobicity, intestinal protein corona formation, as well as holistic properties like particle size, shape, and rigidity. Understanding these properties is essential for enhancing transepithelial transport efficiency and designing effective oral drug delivery systems. Therefore, this review provides a comprehensive overview of the surface and holistic properties that influence the transepithelial transport of nanoparticles, elucidating the underlying principles governing their impact on transepithelial transport. The review also outlines the chosen of parameters to be considered for the subsequent design of oral drug delivery systems.

Within class II bacteriocins, we assume the presence of a separate subfamily of antimicrobial peptides possessing a broad spectrum of antimicrobial activity. Although these peptides are structurally related to the subclass IIa (pediocin-like) bacteriocins, they have significant differences in biological activities and, probably, a mechanism of their antimicrobial action. A representative of this subfamily is acidocin A from Lactobacillus acidophilus TK9201. We discovered the similarity between acidocin A and acidocin 8912 from Lactobacillus acidophilus TK8912 when analyzing plasmids from lactic acid bacteria and suggested the presence of a single evolutionary predecessor of these peptides. We obtained the C-terminally extended homolog of acidocin 8912, named acidocin 8912A, a possible intermediate form in the evolution of the former. The study of secondary structures and biological activities of these peptides showed their structural similarity to acidocin A; however, the antimicrobial activities of acidocin 8912 and acidocin 8912A were lower than that of acidocin A. In addition, these peptides demonstrated stronger cytotoxic and membranotropic effects. Building upon what we previously discovered about the immunomodulatory properties of acidocin A, we studied its proteolytic stability under conditions simulating those in the digestive tract and also assessed its ability to permeate intestinal epithelium using the Caco-2 cells monolayer model. In addition, we found a pronounced effect of acidocin A against fungi of the genus Candida, which might also expand the therapeutic potential of this bacterial antimicrobial peptide. [ABSTRACT FROM AUTHOR]

Despite the promising prospects of nanoparticles in oral drug delivery, the process of oral administration involves a complex transportation pathway that includes cellular uptake, intracellular trafficking, and exocytosis by intestinal epithelial cells, which are necessary steps for nanoparticles to enter the bloodstream and exert therapeutic effects. Current researchers have identified several crucial factors that regulate the interaction between nanoparticles and intestinal epithelial cells, including surface properties such as ligand modification, surface charge, hydrophilicity/hydrophobicity, intestinal protein corona formation, as well as holistic properties like particle size, shape, and rigidity. Understanding these properties is essential for enhancing transepithelial transport efficiency and designing effective oral drug delivery systems. Therefore, this review provides a comprehensive overview of the surface and holistic properties that influence the transepithelial transport of nanoparticles, elucidating the underlying principles governing their impact on transepithelial transport. The review also outlines the chosen of parameters to be considered for the subsequent design of oral drug delivery systems. This review summarizes the crucial factors that influence the transepithelial transport of nanoparticles from the perspective of surface and holistic property modulation. [Display omitted] [ABSTRACT FROM AUTHOR]

Transport of oral nanocarriers across the GI epithelium necessitates transport across hydrophilic mucus layer and the hydrophobic epithelium. Based on hydrophobic-hydrophilic balance, Curcumin-Lipomer (lipid-polymer hybrid nanoparticles) comprising hydrophobic stearic acid and hydrophilic Gantrez™ AN 119 (Gantrez) were developed, by a radical in-situ approach, to successfully traverse both barriers. A monophasic preconcentrate (Cur-Pre) comprising Cur (Curcumin), stearic acid, Gantrez and stabilizers, prepared by simple solution, was added to an aqueous phase to instantaneously generate Curcumin-Lipomer (Cur-Lipo) of nanosize and high entrapment efficiency (EE). Cur-Lipo size and EE was optimized by Box-Behnken Design. Cur-Lipomers of varying hydrophobic-hydrophilic property obtained by varying the stearic acid: Gantrez ratio exhibited size in the range 200–400 nm, EE > 95% and spherical morphology as seen in the TEM. A decrease in contact angle and in mucus interaction, evident with increase in Gantrez concentration, indicated an inverse corelation with hydrophilicity, while a linear corelation was observed for mucopenetration and hydrophilicity. Cur-SLN (solid lipid nanoparticles) which served as the hydrophobic reference revealed contact angle > 90°, maximum interaction with mucus and minimal mucopenetration. The ex-vivo permeation study through chicken ileum, revealed maximum permeation with Cur-Lipo1 and comparable and significantly lower permeation of Cur-Lipo1-D and Cur-SLN proposing the importance of balancing the hydrophobic-hydrophilic property of the nanoparticles. A 1.78-fold enhancement in flux of hydrophobic Cur-SLN, with no significant change in permeation of the hydrophilic Cur-Lipomers (p > 0.05) following stripping off the mucosal layer was observed. This reiterated the significance of hydrophobic-hydrophilic balance as a promising strategy to design nanoformulations with superior permeation across the GI barrier. [ABSTRACT FROM AUTHOR]

Summary: After abalone muscle hot water extract (AHE) was digested in‐vitro, three components (F1, F2, and F3) were separated using the Sephadex G‐15 column, and the antioxidant activity and absorption across Caco‐2 cells were analysed. The result showed that the half‐maximal inhibitory concentrations of F2 for scavenging hydroxyl and ABTS radicals were 0.763 and 0.028 mg mL−1, respectively, which were lower than those of F1 and F3. Moreover, the Trolox equivalent antioxidant capacity was increased after the F2 was transported through the Caco‐2 cell‐monolayered Transwell chamber. The mass spectrometry analysis revealed that the transported samples contained oligopeptides (Leu‐Tyr, Gly‐Ala‐Ala, and His‐Gly‐Ser‐Ala), and the Leu‐Tyr displayed the highest antioxidant activity and transport rate. Overall, these results suggest that oligopeptides with high antioxidant capacities can be produced from AHE through a combination of in‐vitro digestion and absorption across Caco‐2 cells. [ABSTRACT FROM AUTHOR]

7,8‐dihydroxyflavone (7,8‐DHF) is a biologically active flavone with various physiological activities, including neuroprotection, anti‐inflammation, and weight loss. Previous studies have found that the efflux protein P‐glycoprotein (P‐gp) significantly affects the transepithelial transport of 7,8‐DHF in the intestine, resulting in its low oral bioavailability. Based on this, in this study, a Caco‐2 monolayer cell model was used to investigate 14 dietary plant flavonoids as potential P‐gp inhibitors, and their effects on the transepithelial transport and in vitro digestion of 7,8‐DHF were explored. The results showed that among the 14 plant flavonoids, hesperetin, epigallocatechin gallate, fisetin, kaempferol, quercetin, and isoorientin increased and the apparent permeability coefficients (Papp) of 7,8‐DHF at AP → BL direction and lowered Papp value at BL → AP direction to varying degrees, reducing the efflux ratio of 7,8‐DHF less than 1.5. In particular, kaempferol and quercetin exhibited the best effect on promoting the transepithelial transport of 7,8‐DHF, especially when used at molar concentration ratios of 1:1 and 1:2 with 7,8‐DHF. This is beneficial for improving the oral bioavailability of 7,8‐DHF. Meanwhile, 7,8‐DHF was found to maintain structural stability in simulated saliva, gastric juice, and intestinal juice, and its stability was not affected by the coexistence of quercetin and kaempferol. Overall, this study provided a theoretical basis for seeking natural and safe P‐gp inhibitors to improve the oral absorption of natural products. [ABSTRACT FROM AUTHOR]

Liver cirrhosis is associated to circulatory abnormalities leading to hypovolemia and stimulation of the renin-angiotensin-aldosterone system (RAAS). Advanced stages of the disease cause renal failure, impairing K+ and Na+ homeostasis. It has been proposed that the distal colon undergoes functional remodeling during renal failure, in particular by aldosterone-driven increased K+ excretion. In this study, we compared the transcriptional response of aldosterone target genes in the rat distal colon under two models of increased circulating aldosterone (one with concomitant RAAS activation) and in a model of secondary hyperaldosteronism induced by cirrhosis. The expression of a subset of these genes was also tested in distal colon biopsies from control subjects or patients with cirrhosis with varying levels of disease progression and treated or not with mineralocorticoid receptor inhibitor spironolactone. We examined known aldosterone-regulated transcripts involved in corticosteroid signaling and transepithelial ion transport. In addition, we included aldosterone-regulated genes related to cell proliferation. Our comparison revealed multiple aldosterone target genes upregulated in the rat distal colon during decompensated cirrhosis. Epithelial Na+ channel β and γ subunit expression correlated positively with plasma aldosterone concentration and negatively with glomerular filtration rate. Patients with cirrhosis showed increased expression of 11-β-hydroxysteroid-dehydrogenase 2 (11βHSD2), which was reverted by spironolactone treatment, suggesting a sensitization of the distal colon to aldosterone action. In summary, our data show that decaying kidney function during cirrhosis progression toward a decompensated state with hypovolemia correlates with remodeling of distal colon ion transporter expression, supporting a role for aldosterone in the process. [ABSTRACT FROM AUTHOR]

Protein corona (PC) has been identified to impede the transportation of intravenously injected nanoparticles (NPs) from blood circulation to their targeted sites. However, how intestinal PC (IPC) affects the delivery of orally administered NPs are still needed to be elucidated. Here, we found that IPC exerted "positive effect" or "negative effect" depending on different pathological conditions in the gastrointestinal tract. We prepared polystyrene nanoparticles (PS) adsorbed with different IPC derived from the intestinal tract of healthy, diabetic, and colitis rats (H-IPC@PS, D-IPC@PS, C-IPC@PS). Proteomics analysis revealed that, compared with healthy IPC, the two disease-specific IPC consisted of a higher proportion of proteins that were closely correlated with transepithelial transport across the intestine. Consequently, both D-IPC@PS and C-IPC@PS mainly exploited the recycling endosome and ER-Golgi mediated secretory routes for intracellular trafficking, which increased the transcytosis from the epithelium. Together, disease-specific IPC endowed NPs with higher intestinal absorption. D-IPC@PS posed "positive effect" on intestinal absorption into blood circulation for diabetic therapy. Conversely, C-IPC@PS had "negative effect" on colitis treatment because of unfavorable absorption in the intestine before arriving colon. These results imply that different or even opposite strategies to modulate the disease-specific IPC need to be adopted for oral nanomedicine in the treatment of variable diseases. The transepithelial transport-related proteins up-regulated within two disease-specific intestinal PC (IPC), which improved the transcytosis by exploiting the secretory pathways. Consequently, the disease-specific IPC endowed nanoparticles (NPs) with enhanced oral absorption. [Display omitted] [ABSTRACT FROM AUTHOR]

Bread crust constitutes an important by-product of the bakery industry, and its utilization for the isolation of melanoidins to be used as a functional ingredient can enhance its added value and contribute to health. The aim of this study was to evaluate the bioaccessibility, bioactivity, and genoprotective effect of melanoidins derived from bread crust. Bioaccessibility was assessed in gastric, intestinal digestion, and colonic fermentation fractions. The results revealed a relationship between bioaccessible melanoidins and their type (common or soft bread). No cytotoxicity effects were observed for bioaccessible fractions, as assessed by MTT and RTA methods, and they did not affect the distribution of E-cadherin in Caco-2 cells, confirming their ability to maintain membrane integrity. Furthermore, our study demonstrated that the gastrointestinal and colonic fermentation fractions successfully transported across the intestinal barrier, without affecting cell permeability, and showed antioxidant activity on the basolateral side of the cell monolayer. Remarkably, both fractions displayed a significant genoprotective effect in Caco-2 cells. Our findings provide crucial insights into the relationship between the melanoidins and their bioactivity and genoprotective effect. These results demonstrated the potential of bioaccessible melanoidins as valuable bioactive compounds for the development of functional foods, without showing toxic effects on gastrointestinal cells. [ABSTRACT FROM AUTHOR]

The concept of solvent drag, i.e., water and solutes sharing the same pore and their transport being frictionally coupled, was first proposed in the early 1950s. During the following decades, it was applied to transport processes across cell membranes as well as transport along the paracellular pathway. Water-driven solute transport was proposed as the major mechanism for electrolyte and nutrient absorption in the small intestine and for Cl− and HCO3− reabsorption in the renal proximal tubule. With the discovery of aquaporins as transcellular route for water transport and the claudin protein family as the major determinant of paracellular transport properties, new mechanistic insights in transepithelial water and solute transport are emerging and call for a reassessment of the solvent drag concept. Current knowledge does not provide a molecular basis for relevant solvent drag-driven, paracellular nutrient, and inorganic anion (re-)absorption. For inorganic cation transport, in contrast, solvent drag along claudin-2-formed paracellular channels appears feasible. [ABSTRACT FROM AUTHOR]

Studying the composition of a certain food is not enough to predict its health benefits. Research over the past decades has decisively strengthened the notion that any putative health benefit is best related to the fraction of compounds transferred from ingested foods into the body since the absorption may be incomplete after oral consumption. In other words, the bioavailability of food components is crucial information. Therefore, a variety of in vitro models have been developed to predict their bioaccessibility and bioavailability in the most diverse food matrices and food products. These models can also be applied to study the impact of several endogenous or exogenous factors on the bioaccessibility and bioavailability of nutrients and bioactive compounds, guiding nutrition and food scientists, technologists, and engineers towards the development of strategies to optimize the positive impact of the diet on well-being and quality of life. While bioavailability is ideally examined in human volunteers, in vitro digestion methods, as well as intestinal absorption and microphysiological models, simulate human physiological conditions. Additionally, in vitro methods are alternatives to offset ethical, economical, and experimental limitations associated with in vivo studies conducted either with individuals or animals. This graphical review draws parallels between in vitro models mimicking digestion processes, uptake, absorption, metabolism, and distribution of dietary compounds and human physiology.

Phycotoxins are a class of multiple natural metabolites produced by microalgae in marine and freshwater ecosystems that bioaccumulate in food webs, particularly in shellfish, having a great impact on human health. Phycotoxins are mainly leached and absorbed in the small intestine when human consumers accidentally ingest toxic aquatic products contaminated by them. To assess the intestinal uptake and damage of phycotoxins, a typical in vitro model was developed and widely applied using the human colorectal adenocarcinoma Caco-2 cell line. In this review, the application cases were summarized for multiple phycotoxins, including microcystins (MCs), cylindrospermopsins (CYNs), domoic acids (DAs), saxitoxins (STXs), palytoxins (PLTXs), okadaic acids (OAs), pectenotoxins (PTXs) and azaspiracids (AZAs). The results of the previous studies showed that each group of phycotoxins presented different cytotoxicity and mechanisms to Caco-2 cells, and significant discrepancies in the transport of phycotoxin across the Caco-2 cell monolayers. Therefore, this review describes the evaluation assays of the Caco-2 cell monolayer model, illustrates the principles of several primary cytotoxicity evaluation assays, and summarizes the cytotoxicity of each group of phycotoxins to Caco-2 cells line and their cellular transport, and finally proposes the development of multicellular intestinal models for future comprehensive studies on the toxicity and absorption of phycotoxins in the intestine. It will improve the understanding of Caco-2 cell monolayer models in the toxicology studies on phycotoxins and the potentially detrimental effects of microalgal toxins on the human intestine.

Bread crust constitutes an important by-product of the bakery industry, and its utilization for the isolation of melanoidins to be used as a functional ingredient can enhance its added value and contribute to health. The aim of this study was to evaluate the bioaccessibility, bioactivity, and genoprotective effect of melanoidins derived from bread crust. Bioaccessibility was assessed in gastric, intestinal digestion, and colonic fermentation fractions. The results revealed a relationship between bioaccessible melanoidins and their type (common or soft bread). No cytotoxicity effects were observed for bioaccessible fractions, as assessed by MTT and RTA methods, and they did not affect the distribution of E-cadherin in Caco-2 cells, confirming their ability to maintain membrane integrity. Furthermore, our study demonstrated that the gastrointestinal and colonic fermentation fractions successfully transported across the intestinal barrier, without affecting cell permeability, and showed antioxidant activity on the basolateral side of the cell monolayer. Remarkably, both fractions displayed a significant genoprotective effect in Caco-2 cells. Our findings provide crucial insights into the relationship between the melanoidins and their bioactivity and genoprotective effect. These results demonstrated the potential of bioaccessible melanoidins as valuable bioactive compounds for the development of functional foods, without showing toxic effects on gastrointestinal cells.

Considering the involvement of oxidative stress in the etiology of many non-communicable diseases, food-derived antioxidant peptides (FDAPs) are strong candidates for nutraceutical development for disease prevention and management. This paper reviews current evidence on the transepithelial transport and cellular mechanisms of antioxidant activities of FDAPs. Several FDAPs have multiple health benefits such as anti-inflammatory and anti-photoaging activities, in addition to antioxidant properties through which they protect cellular components from oxidative damage. Some FDAPs have been shown to permeate the intestinal epithelium, which could facilitate their bioavailability and physiological bioactivities. Molecular mechanisms of FDAPs include suppression of oxidative stress as evidenced by reduction in intracellular reactive oxygen species production, lipid peroxidation and apoptotic protein activation as well as increase in antioxidant defense mechanisms (enzymatic and non-enzymatic). Since many FDAPs have demonstrated promising antioxidant activity, future investigation should focus on further elucidation of molecular mechanisms and human studies to explore their practical application for the prevention and management of oxidative stress-related diseases.

[Display omitted] • The intestinal absorption of casein peptide Met-Lys-Pro (MKP) was studied in vitro. • Transepithelial transport of MKP in casein hydrolysate exceeded that of MKP alone. • The peptide matrix in casein hydrolysate may suppress MKP degradation. High blood pressure is a major risk factor for cardiovascular disease. Our previous study confirmed that daily intake of casein hydrolysate that contained Met-Lys-Pro (MKP) can safely lower mildly elevated blood pressure. The present study aimed to evaluate the intestinal absorption differences between peptide MKP as a casein hydrolysate and synthetic MKP alone using Caco-2 cells and human iPS cell-derived small intestinal epithelial cells (hiSIECs). MKP was transported intact through Caco-2 cells and hiSIECs with permeability coefficient (P app) values of 0.57 ± 0.14 × 10-7 and 1.03 ± 0.44 × 10-7 cm/s, respectively. This difference in P app suggests differences in the tight junction strength and peptidase activity of each cell. Moreover, the transepithelial transport and residual ratio of intact MKP after adding casein hydrolysate containing MKP was significantly higher than that after adding synthetic MKP alone, suggesting that other peptides in casein hydrolysate suppressed MKP degradation and increased its transport. These findings suggest that hiSIECs could be useful for predicting the human intestinal absorption of bioactive peptides; ingesting MKP as a casein hydrolysate may also improve MKP bioavailability. [ABSTRACT FROM AUTHOR]

The choroid plexus epithelium (CPe) forms a barrier between the cerebral blood supply and the cerebrospinal fluid (CSF), establishing the blood-CSF barrier (BCSFB). CSF is actively secreted by the CPe via tightly controlled processes involving multiple channels, transporters, and pumps. The importance of controlling CSF production and composition has been accentuated recently with an appreciation of CSF dysfunction in many pathologies. For mechanistic studies of CSF production, isolated CPe cell lines are valuable for the testing of hypotheses and potential drug targets. Although several continuous CPe cell lines have been described, none appear to have all the characteristics of the native epithelium and each must be used judiciously. The porcine choroid plexus-Riems (PCP-R) cell line forms a high-resistance monolayer characteristic of a barrier epithelium. Conservation of this phenotype is unusual among CPe cell lines, making this model useful for studies of the effects of infection, injury, and drugs on permeability. We have recently discovered that, although this line expresses many of the transporters expressed in the native tissue, some are mispolarized. As a result, inferences regarding fluid/electrolyte flux and the resultant CSF production should be pursued with caution. Furthermore, extended culture periods and changes in media composition result in significant morphological and functional variability. These studies provide a more detailed characterization of the PCP-R cell line concerning transporter expression, polarization, and functionality, as well as plasticity in culture, with the goal to provide the scientific community with information necessary to optimize future experiments with this model. [ABSTRACT FROM AUTHOR]

In this study, the in vitro bioavailability of rosehip infusion phenolics, mainly catechin, as a response to conventional and non-thermal treatments by combining gastrointestinal digestion and a Caco-2 cell culture model, was investigated. After application of thermal treatment (TT, 85 °C/10 min), high pressure (HPP, 600 MPa/5 min) or pulsed electric field (PEF, 15 kJ/kg) processing, all samples were subjected to simulated gastrointestinal digestion. Then, the amount of maximum non-toxic digest ratio was determined by the cytotoxicity sulforhodamine B (SRB) assay. Next, Caco-2 cells were exposed to 1:5 (v/v) times diluted digests in order to simulate the transepithelial transportation of catechin. Results showed that non-thermally processed samples (5.19 and 4.62% for HPP and PEF, respectively) exhibited greater transportation across the epithelial cell layer compared to than that of the TT-treated sample (3.42%). The present study highlighted that HPP and PEF, as non-thermal treatments at optimized conditions for infusions or beverages, can be utilized in order to enhance the nutritional quality of the final products. [ABSTRACT FROM AUTHOR]

Summary: In this study, a promising nanoparticle of casein peptide–zinc chelate (CNP‐Zn) for zinc supplementation was obtained. The structure properties and transepithelial transport mechanism were investigated. The amino acid composition, particle size, zeta potential and microstructure were significantly changed after chelation with zinc ions. The results of fourier‐transform infrared spectroscopy suggested that the oxygen atoms from both the carboxyl and carbonyl groups, amides, were responsible for zinc chelation. After gastrointestinal digestion, over 40% of zinc remained bound in CNP–Zn. The transepithelial transport results showed that paracellular route and PepT1 transporter were involved in the absorption of CNP–Zn, and almost 50% of the bound zinc was co‐transported with peptides. These findings would provide a relatively comprehensive cognition for developing efficient dietary zinc carriers. [ABSTRACT FROM AUTHOR]

Purpose: Bevacizumab is taken up and transported through the retinal pigment epithelium. Inflammatory signaling may influence this interaction. In the present study, we have investigated the effect of pro-inflammatory stimuli on the uptake, intracellular localization, and transepithelial transport of bevacizumab. Methods: ARPE-19 cell line or primary porcine RPE cells were treated with clinical relevant concentrations of bevacizumab (250 µg/ml). Pro-inflammatory signaling was induced by TLR-3 agonist polyinosinic:polycytidylic acid (Poly I:C). Viability was investigated with MTT and trypan-blue exclusion assay, and cell number, uptake, and intracellular localization were investigated with immunofluorescence, investigating also actin filaments, the motor protein myosin 7a and lysosomes. Immunofluorescence signals were quantified. Intracellular bevacizumab was additionally detected in Western blot. Barrier function was investigated with transepithelial resistant measurements (TER). The transepithelial transport of bevacizumab and its influence on cytokine (IL-6, IL-8, IL-1β, TNFα) secretion was investigated with ELISA. Results: Poly I:C in combination with bevacizumab reduced the viability of the cells. Treatment with Poly I:C reduced the uptake of bevacizumab, changed the intensity of the actin filaments, and reduced the colocalization with myosin 7a. In addition, Poly I:C reduced the capacity of RPE cells to transport bevacizumab over the barrier. In addition, bevacizumab reduced the secretion of IL-8 and TNFα after Poly I:C stimulation at selected time points. Conclusions: Pro-inflammatory activation of RPE cells with TLR-3 agonist Poly I:C changes the interaction of RPE cells with the anti-VEGF compound bevacizumab, reducing its uptake and transport. On the other hand, bevacizumab might influence pro-inflammatory cytokine release. Our data indicate that inflammation may influence the pharmacokinetic of bevacizumab in the retina. [ABSTRACT FROM AUTHOR]

Zuxian Chen,1,* Shangcong Han,1,* Xiaotang Yang,1 Lisa Xu,1 Hong Qi,2 Guizhou Hao,3 Jie Cao,1 Yan Liang,1 Qingming Ma,1 Guimin Zhang,3 Yong Sun1 1Department of Pharmaceutics, School of Pharmacy, Qingdao University, Qingdao 266021, People’s Republic of China; 2Department of General Surgery, Qingdao Municipal Hospital, Qingdao 266071, People’s Republic of China; 3Department of Scientific Research, Lunan Pharmaceutical Corporation, Linyi 276001, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yong Sun Tel +86-532-82991203Email sunyong@qdu.edu.cnBackground: Although dynamics and uses of modified nanoparticles (NPs) as orally administered macromolecular drugs have been researched for many years, measures of molecule stability and aspects related to important transport-related mechanisms which have been assessed in vivo remain as relatively under characterized. Thus, our aim was to develop a novel type of oral-based delivery system for insulin and to overcome barriers to studying the stability, transport mechanisms, and efficacy in vivo of the delivery system.Methods: NPs we developed and tested were composed of insulin (INS), dicyandiamide-modified chitosan (DCDA-CS), cell-penetrating octaarginine (r8), and hydrophilic hyaluronic acid (HA) and were physically constructed by electrostatic self-assembly techniques.Results: Compared to free-insulin, levels of HA-DCDA-CS-r8-INS NPs were retained at more desirable measures of biological activity in our study. Further, our assessments of the mechanisms for NPs suggested that there were high measures of cellular uptake that mainly achieved through active transport via lipid rafts and the macropinocytosis pathway. Furthermore, investigations of NPs indicated their involvement in caveolae-mediated transport and in the DCDA-CS-mediated paracellular pathway, which contributed to increasing the efficiency of sequential transportation from the apical to basolateral areas. Accordingly, high efficiency of absorption of NPs in situ for intestinal loop models was realized. Consequently, there was a strong induction of a hypoglycemic effect in diabetic rats of NPs via orally based administrations when compared with measures related to free insulin.Conclusion: Overall, the dynamics underlying and influenced by HA-DCDA-CS-r8-INS may hold great promise for stability of insulin and could help overcome interference by the epithelial barrier, and thus showing a great potential to improve the efficacy of orally related treatments.Keywords: insulin, oral drug delivery, transepithelial transport, paracellular pathway, caveolae-mediated transport

In this study, the in vitro bioavailability of rosehip infusion phenolics, mainly catechin, as a response to conventional and non-thermal treatments by combining gastrointestinal digestion and a Caco-2 cell culture model, was investigated. After application of thermal treatment (TT, 85 °C/10 min), high pressure (HPP, 600 MPa/5 min) or pulsed electric field (PEF, 15 kJ/kg) processing, all samples were subjected to simulated gastrointestinal digestion. Then, the amount of maximum non-toxic digest ratio was determined by the cytotoxicity sulforhodamine B (SRB) assay. Next, Caco-2 cells were exposed to 1:5 (v/v) times diluted digests in order to simulate the transepithelial transportation of catechin. Results showed that non-thermally processed samples (5.19 and 4.62% for HPP and PEF, respectively) exhibited greater transportation across the epithelial cell layer compared to than that of the TT-treated sample (3.42%). The present study highlighted that HPP and PEF, as non-thermal treatments at optimized conditions for infusions or beverages, can be utilized in order to enhance the nutritional quality of the final products.

Understanding the transport mechanism of the peptide Asn-Cys-Trp (NCW) is crucial to improving its intestinal absorption and bioavailability. This study investigated the absorption of NCW through Caco-2 cell monolayers and its interaction with the DPPC bilayers. Results revealed that after a 3 h incubation, the Papp (AP-BL) and Papp (BL-AP) values of NCW at a concentration of 5 mmol/L were (22.24 ± 4.52) × 10 -7 and (6.63 ± 2.31) × 10 -7 cm/s, respectively, with the transport rates of 1.59 ± 0.32 and 0.62 ± 0.20%, indicating its moderate absorption. NCW was found to be transported via PepT1 and paracellular transport pathways, as evidenced by the significant impact of Gly-Pro and cytochalasin D on the Papp values. Moreover, NCW upregulated ZO-1 mRNA expression. Further investigation of the ZO-1-mediated interaction between NCW and tight junction proteins will contribute to a better understanding of the paracellular transport mechanism of NCW. The interaction between NCW and the DPPC bilayers was predominantly driven by entropy. NCW permeated the bilayers through electrostatic, hydrogen bonding, and hydrophobic interactions, resulting in increased fluidity, flexibility, and disorder as well as phase transition and phase separation of the bilayers.

Previous research has shown the absorption of polybrominated diphenyl ethers (PBDEs) in the human gastrointestinal tract, but limited attention has been given to the influence of nutrients on PBDE absorption from food matrices. We investigated the effects of nutrients (oil, starch, protein, and dietary fiber) on the absorption and transport of PBDEs in a Caco-2 cell model and bioaccessibility of PBDEs by an in vitro gastrointestinal digestion method. The results showed that the accumulation ratios of PBDE congeners in Caco-2 cells were higher in the nutrient addition groups (oil: 26.7–50.6%, starch: 27.0–58.7%, protein: 12.1–44.1%, and dietary fiber: 28.2–55.1%) than the control group (7.17–36.1%), whereas the transport ratios were lower (oil: 2.30–7.20%, starch: 1.55–9.15%, protein: 1.04–8.78%, and dietary fiber: 0.85–7.04%) than control group (3.78–11.1%). Additionally, the PBDE bioaccessibility could be increased by adding the nutrients, particularly oil and starch. This study clarified the differences in PBDE absorption in the presence of nutrients using the in vitro digestion and Caco-2 cell model. The findings showed that nutrients were an important factor that promoted PBDE absorption in the gastrointestinal tract. Therefore, it is important to focus on a novel dietary strategy of food consumption with contaminant compounds to protect human health.

Permeability of angiotensin I-converting enzyme (ACE) inhibitory peptides (KPLLCS, ELFTT, and KPLL) identified from the in vitro gastrointestinal digest of cooked chicken breast was investigated using the Caco-2 cell model system. KPLLCS was originally the most effective ACE inhibitor (IC50 0.37 μM), but it was degraded during permeation through Caco-2 cells. Among these peptides, KPLL showed the highest permeability in intact form, and partially degraded to KP and LL, which still showed ACE inhibitory activity after permeation. ACE inhibitory activity of permeated KPLL was highest of 56%. KPLL and KP possessed a mixed inhibitor characteristic, while LL was a non-competitive inhibitor. Based on molecular docking, K at N-terminus of KPLL is a key structure contributing to ACE inhibition as it can occupy the active site of ACE. Determining the structural stability and degree of peptide transport across epithelial cell is required to confirm their potential as ACE inhibitors.

Peptides from Alcalase-hydrolyzed soybean protein hydrolysate (SPH) may hold the potential as natural antioxidants. In addition, the effect of human gastrointestinal (GI) tract on peptide bioavailability needs to be explored. In this study, the impact of simulated GI digestion and transepithelial transport on various antioxidant properties of SPH were investigated. SPH displayed DPPH radical scavenging (IC50 = 4.22 mg/mL), ABTS·+ radical scavenging (IC50 = 2.93 mg/mL), reducing power and metal ion-chelating activities (IC50 = 0.67 mg/mL). Furthermore, SPH significantly (P

The Octapeptide ELHQEEPL, which was identified from the rapeseed protein napin showed prominent Dipeptidyl peptidase‐IV (DPP‐IV) inhibitory activity. The objective of this study was to investigate the DPP‐IV inhibitory activity and transepithelial transport of ELHQEEPL in an approaching intestinal condition using Caco‐2 cell monolayers. ELHQEEPL and its degraded fragments EL, HQEEP, and methylated ELHQEEPL were transported across Caco‐2 cell monolayers through different pathways. Compared with the nonbiological enzyme inhibition test, the in vitro experiment on Caco‐2 cell monolayers showed that the IC50 value of DPP‐IV inhibition increased by 43.11% for ELHQEEPL. There was no significant change in DPP‐IV gene expression in the Caco‐2 cell monolayers upon treatment with ELHQEEPL. Furthermore, molecular docking predicted that the weaker binding between inhibitory peptide and enzyme for the degradation products from ELHQEEPL during transepithelial transport greatly limited its role in inhibiting DPP‐IV. Practical applications: The DPP‐IV inhibitory activity of ELHQEEPL was confirmed using Caco‐2 cell monolayers as a novel assessment tool, although its potency was reduced by metabolic degradation. In general, this study reported the use of Caco‐2 cell monolayers as a tool for comprehensively studying peptides as sources of DPP‐IV inhibitors. A Caco‐2 cell‐based approach with molecular docking can be adapted for the investigation of intestinal absorption and activity attenuation of food peptides being considered for enzymatic action. Moreover, since the Caco‐2 cells express a wide range of enzymes, this method can be used for screening for other active food peptides such as for the inhibitors of ACE and a‐glucosidase. [ABSTRACT FROM AUTHOR]

The objective of this paper was to investigate the effects of hydrophobicity and molecular weight (MW) on the transepithelial transport permeability of oligopeptides across Caco‐2 cell monolayers. Results showed that oligopeptides with different N‐terminal amino acids had a wide range of permeability values and could be divided into three levels according to their correlations with log D and MW. At a good level of permeability, the permeability was positively correlated with log D, but negatively correlated with MW (p <.001); at an intermediate level of permeability, the permeability was negatively correlated with log D and MW (p <.001); and at a low level of permeability, the permeability was positively correlated with log D and MW (p <.01). These results suggest for the first time that the transport of oligopeptides across Caco‐2 cell monolayers might be closely related to their molecular properties of log D and MW. Practical applications: A great number of food‐derived bioactive peptides display health‐promoting effects and show potential as bioactive ingredients in functional foods. However, the poor absorption in the intestine limits the application of food bioactive peptides, especially for the oligopeptides containing more than three amino acids. Although the transepithelial transport of food‐derived oligopeptides in the intestinal epithelium has been widely reported, its transport mechanism is still obscure. Our study shows a three‐level relationship between the transport permeability and log D and MW of oligopeptides across Caco‐2 cell monolayers and provides a novel evidence for the coexistence of transcellular and paracellular pathways for the transport of oligopeptides through the intestine. This result will contribute to the understanding of the transport mechanisms of oligopeptides in the intestine. [ABSTRACT FROM AUTHOR]

Aronia phenolics are considered to be beneficial for cardiovascular health, but are not always bioavailable. In this study, we evaluated the in vitro bioavailability of Aronia juice phenolics by combination of intestinal digestion models with Caco-2 absorption models. First, intestinal luminal stability and microbial metabolism of Aronia phenolics were investigated using two in vitro digestion models: a short-term batch model and a long term dynamic model (SHIME®). Next, diluted digests were directly applied to Caco-2 cells to simulate enterocyte absorption. Three anthocyanins, 5 flavonol glycosides, 5 phenolic acids and epicatechin were quantified at different stages of digestion and absorption. The location of absorption was compound specific, as procyanidins were mainly absorbed in the distal colon while anthocyanins, flavonol glycosides and phenolic acids were absorbed in the proximal intestinal tract. Further, the food matrix present in the colon significantly increased transport without permanent barrier damage.

Recently, due to their promising applications in biomedicine, magnetic iron oxide nanoparticles (MPs) have become one of the research hotspots in the nanomedicine field. Since various synthetic modifications have been widely applied to these nanoparticles for better targeting behaviors, it is meaningful to apply the optimal magnetic field condition for each case. This will enable creating a safe and efficient drug targeting using different types of MPs. In the present study, we aimed to find out any changes of transepithelial transport of polysaccharide-coated MPs by applying the continuous or the pulsatile magnetic field condition. Our results with heparin-functionalized MPs indicate that the particle concentrations and the external magnetic field could influence the transepithelial permeability of the particles. In the presence of a continuously applied magnetic density, heparin-MPs at high concentrations, by forming magnetically-induced aggregation of particles over the cell surface layer, showed a lower cellular transport than those at low concentrations. Furthermore, the results from the quantitative chemical assays and imaging analyses showed that transepithelial transport of heparin-MPs (negatively charged) under the pulsatile magnetic field was higher than that under the continuous magnetic field (CP), whereas the starch-MPs (neutrally charged) showed a small difference in transepithelial transport or cell retention between pulsatile vs. continuous magnetic field conditions. Taken together, our results suggest that the external magnetic field should be differentially applied to control the cellular drug transport depending on the physicochemical properties of the surface chemistry of magnetic particles. [ABSTRACT FROM AUTHOR]

In recent years, food-derived peptides have gained much attention for their potential health benefits. Some short and medium-sized peptides released from food proteins after their enzymatic hydrolysis may exhibit hypocholesterolemic activity. Hypocholesterolemic peptides act either by targeting exogenous cholesterol in the gastrointestinal (GI) tract or by modulating endogenous cholesterol levels via cholesterol metabolism pathways in the liver after being absorbed. This paper provides a comprehensive review of current pieces of evidence regarding the production, transepithelial transport, and cellular mechanisms underlying the hypocholesterolemic activities of food-derived peptides. The molecular mechanisms of hypocholesterolemic peptides involve bile acid binding, inhibition of cholesterol micellar solubility, statin-like effects through the modulation of 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGCoAR), as well as the targeting of interactions between proprotein convertase subtilisin/kexin type 9 (PCSK9) and low-density lipoprotein receptor (LDLR), sterol regulatory element-binding protein 2 (SREBP-2), and hepatocyte nuclear factor 1α (HNF-1α) pathways. Furthermore, some peptides exhibit multiple biological activities, such as anti-inflammatory and antioxidant activities, besides cholesterol-lowering properties, thereby safeguarding cellular components against high levels of cholesterol-induced damage. However, since only a few studies have evaluated the in vivo effects of hypocholesterolemic peptides, further studies carried out in animal models or human are necessary to exploit these ingredients in the prevention and management of hypercholesterolemia. • Some peptides exert their activity through the modulation of HMGCoAR. • Some peptides inhibit the interactions between PCSK9 and LDL receptor. • They may be used in the prevention of hypercholesterolemia. [ABSTRACT FROM AUTHOR]

We hypothesized that, due to the high pH of this compartment, the reticulum epithelium displays particular features in the transport of short-chain fatty acids (SCFA). Ovine reticulum epithelium was incubated in Ussing chambers using a bicarbonate-free buffer solution containing butyrate (20 mmol L−1). p-hydroxymercuribenzoic acid (pHMB), 5-(N-Ethyl-N-isopropyl)amiloride (EIPA), or ouabain were added to the buffer solution as inhibitors of monocarboxylate transporters, sodium-proton-exchangers, or the Na+/K+-ATPase, respectively. The short-circuit current (Isc) and transepithelial conductance (Gt) were monitored continuously while the flux rates of 14C-labelled butyrate were measured in the mucosal-to-serosal (Jmsbut) or serosal-to-mucosal direction (Jsmbut). Under control conditions, the mean values of Isc and Gt amounted to 2.54 ± 0.46 µEq cm−2 h−1 and 6.02 ± 3.3 mS cm−2, respectively. Jmsbut was 2.1 ± 1.01 µmol cm−2 h−1 on average and about twice as high as Jsmbut. Incubation with ouabain reduced Jmsbut, while Jsmbut was not affected. The serosal addition of EIPA did not affect Jmsbut but reduced Jsmbut by about 10%. The addition of pHMB to the mucosal or serosal solution reduced Jmsbut but had no effect on Jsmbut. Mucosally applied pHMB provoked a transient increase in the Isc. The serosal pHMB sharply reduced Isc. Our results demonstrate that butyrate can be effectively transported across the reticulum epithelium. The mechanisms involved in this absorption differ from those known from the rumen epithelium.

The epididymis is a single convoluted tubule lined by a pseudostratified epithelium. Specialized epididymal epithelial cells, the so-called principal, basal, narrow, and clear cells, establish a unique luminal environment for the maturation and storage of spermatozoa. The epididymis is functionally and structurally divided into several segments and sub-segments that create regionally distinct luminal environments. This organ is immature at birth, and epithelial cells acquire their fully differentiated phenotype during an extended postnatal period, but the factors involved in this complex process remain incompletely characterized. In the adult epididymis, the establishment of an acidic luminal pH and low bicarbonate concentration in the epididymis contributes to preventing premature activation of spermatozoa during their maturation and storage. Clear cells are proton-secreting cells throughout the epididymis, but principal cells have distinct acid/base transport properties, depending on their localization within the epididymis. Basal cells are located in all epididymal segments, but they have a distinct morphology depending on the segment and species examined. How this structural plasticity of basal cells is regulated is discussed here. Also, the role of luminal factors and androgens in the regulation of epithelial cells is reviewed in relation to their respective localization in the proximal versus distal regions of the epididymis. Finally, we describe a novel role for CFTR in tubulogenesis and epithelial cell differentiation.

In order to evaluate potential therapeutic properties of Sambucus nigra fruit extract, its anti-inflammatory effects were investigated in lipopolysaccharide-activated RAW264.7 macrophages. Physiochemical changes in the extract composition, which occur following oral administration, were mimicked in an artificial alimentary tract, while transepithelial absorption was simulated using enterocyte-like Caco-2 cell monolayer, imitating intestinal barrier. The obtained results showed that gastrointestinal digested extract, transported across intestinal epithelium, down-regulated the expression of major genes of inflammatory pathway, such as IL-1β, IL-6, TNF-α and COX-2. Reduction of IL-6, TNF-α and prostaglandin E2 secretion was also observed as a result of LPS-stimulated macrophages' treatment. Moreover, enhanced nitric oxide production in response to inflammatory stimuli in RAW264.7 cells was controlled by the digested and intestinal permeable fraction of elderberry extract.

The effect of a chum salmon egg lectin (CSL3) on tight junction (TJ) of Caco-2 cell monolayers was investigated. The lectin opened TJ as indicated by the decrease of the transepithelial electrical resistance (TER) value and the increase of the permeation of lucifer yellow, which is transported via the TJ-mediated paracellular pathway. The effects of CSL3 were inhibited by the addition of 10 mM L-rhamnose or D-galactose which were specific sugars for CSL3. The lectin increased the intracellular Ca2+ of Caco-2 cell monolayers, that could be inhibited by the addition of L-rhamnose. The fluorescence immunostaining of β-actin in Caco-2 cell monolayers revealed that the cytoskeleton was changed by the CSL3 treatment, suggesting that CSL3 depolymerized β-actin to cause reversible TJ structural and functional disruption. Although Japanese jack bean lectin and wheat germ lectin showed similar effects in the decrease of the TER values and the increase of the intracellular Ca2+, they could not be inhibited by the same concentrations of simple sugars, such as D-glucose and N-acetyl-D-glucosamine.

Assessing nutrient bioavailability is complex, as the process involves multiple digestion steps, several cellular environments, and regulatory-metabolic mechanisms. Several in vitro models of different physiological relevance are used to study nutrient absorption, providing significant challenges in data evaluation. However, such in vitro models are needed for mechanistic studies as well as to screen for biological functionality of the food structures designed. This collaborative work aims to put into perspective the wide-range of models to assay the permeability of food compounds considering the particular nature of the different molecules, and, where possible, in vivo data are provided for comparison.

Proximal tubules in the kidney play a crucial role in reabsorbing and eliminating substrates from the body into the urine, leading to high local concentrations of xenobiotics. This makes the proximal tubule a major target for drug toxicity that needs to be evaluated during the drug development process. Here, we describe an advanced in vitro model consisting of fully polarized renal proximal tubular epithelial cells cultured in a microfluidic system. Up to 40 leak-tight tubules were cultured on this platform that provides access to the basolateral as well as the apical side of the epithelial cells. Exposure to the nephrotoxicant cisplatin caused a dose-dependent disruption of the epithelial barrier, a decrease in viability, an increase in effluent LDH activity, and changes in expression of tight-junction marker zonaoccludence 1, actin, and DNA-damage marker H2A.X, as detected by immunostaining. Activity and inhibition of the efflux pumps P-glycoprotein (P-gp) and multidrug resistance protein (MRP) were demonstrated using fluorescence-based transporter assays. In addition, the transepithelial transport function from the basolateral to the apical side of the proximal tubule was studied. The apparent permeability of the fluorescent P-gp substrate rhodamine 123 was decreased by 35% by co-incubation with cyclosporin A. Furthermore, the activity of the glucose transporter SGLT2 was demonstrated using the fluorescent glucose analog 6- NBDG which was sensitive to inhibition by phlorizin. Our results demonstrate that we developed a functional 3D perfused proximal tubule model with advanced renal epithelial characteristics that can be used for drug screening studies. [ABSTRACT FROM AUTHOR]

Korat-chicken breast and thigh were subjected to heating at 70, 100 or 121 °C for 30 min and simulated in vitro gastrointestinal digestion. At 70 or 100 °C heating, digests of breast possessed higher ACE inhibitory activity than those of thigh. The highest ACE inhibitory activity was found in the digest of breast cooked at 70 °C (B/H-70), whereas breast heated at 121 °C (B/H-121) exhibited the lowest. The 1-kDa permeate of the B/H-70 digest revealed higher permeability through colorectal adenocarcinoma monolayers and ACE inhibitory activity than did B/H-121. Among nine transported peptides, APP derived from myosin showed the highest ACE inhibition, with a non-competitive characteristic (K i 0.93 μM). Molecular docking showed that APP interacts with ACE via hydrogen bonds, electrostatic and van der Waals interactions. In conclusion, mild thermal treatment of chicken breast resulted in a higher amount of transported peptides, exerting higher ACE inhibitory activity, which could lead to potential health benefits. [ABSTRACT FROM AUTHOR]

Alpha-lipoic acid (ALA) oral supplements were used in many pathologies associated with increased oxidative stress. Although only R -ALA is considered the biologically active form, R , S -ALA is used in therapeutic applications even showing poor water solubility. The aim of this work was to study the absorption and transport mechanism across the intestinal barrier of new R -ALA stable and water soluble form, consisting in the lysine R -ALA salt, in presence and absence of specific inhibitors of Na + /multivitamin (SMVT) and monocarboxylic acids (MCT). The absorption of a new ALA form was investigated at rat everted sacs in comparison with R -ALA, S -ALA, and R , S -ALA. Results showed that duodenum is the best portion of intestine for ALA forms absorption. The absorption percentage of R -ALA, S -ALA, R , S -ALA, and lysine R -ALA salt was 66%, 43%, 55%, and 70%, respectively. The modest effect of the SMVT inhibitor biotin demonstrated that this transporter system is not principally involved in the absorption of lysine R -lipoate salt across the rat intestinal barrier. On the contrary, the MCT inhibitor octanoic acid significantly reduced the transport of this salt, whit an absorption decrease of R -ALA and lysine R -lipoate salt of 28% and 24%, respectively. Since the highest concentration of these inhibitors did not completely inhibit the absorption of lysine R -lipoate salt, other transport mechanisms probably operate for its intracellular delivery. The new form of ALA, lysine R -lipoate salt, was the most absorbed respect to the other ALA forms demonstrating that this compound is more suitable for oral administration. This new salt could represent a promising candidate for ALA oral supplementation. [ABSTRACT FROM AUTHOR]

Permeability of angiotensin I-converting enzyme (ACE) inhibitory peptides (KPLLCS, ELFTT, and KPLL) identified from the in vitro gastrointestinal digest of cooked chicken breast was investigated using the Caco-2 cell model system. KPLLCS was originally the most effective ACE inhibitor (IC 50 0.37 μM), but it was degraded during permeation through Caco-2 cells. Among these peptides, KPLL showed the highest permeability in intact form, and partially degraded to KP and LL, which still showed ACE inhibitory activity after permeation. ACE inhibitory activity of permeated KPLL was highest of 56%. KPLL and KP possessed a mixed inhibitor characteristic, while LL was a non-competitive inhibitor. Based on molecular docking, K at N-terminus of KPLL is a key structure contributing to ACE inhibition as it can occupy the active site of ACE. Determining the structural stability and degree of peptide transport across epithelial cell is required to confirm their potential as ACE inhibitors. [ABSTRACT FROM AUTHOR]

Antioxidant casein peptides were separated by SP-Sephadex C-25 and C 18 columns. The transepithelial transport and bioavailability including the transport ratio and the remaining ratios of antioxidant activity (RRAA) of these peptide absorbates, were then investigated using a Caco-2 cell monolayer. The results indicate that both the negatively charged peptide fractions (CF1, CF2 and CF3) and the more hydrophilic fraction (HF1) were mainly transported via PepT1 and paracellular routes. The positively charged fractions (CF4 and CF5) and hydrophobic fractions (HF2, HF3 and HF4) were transported via PepT1 and transcytosis. The strongly negatively charged and more hydrophobic fractions showed a higher transport ratio, which ranged from 9.5 to 12.5%; however, the transport ratio of positively charged and hydrophilic fractions ranged from 4.0 to 8.5%. The positively charged and hydrophilic fractions showed a higher RRAA. [ABSTRACT FROM AUTHOR]

Peptides from Alcalase-hydrolyzed soybean protein hydrolysate (SPH) may hold the potential as natural antioxidants. In addition, the effect of human gastrointestinal (GI) tract on peptide bioavailability needs to be explored. In this study, the impact of simulated GI digestion and transepithelial transport on various antioxidant properties of SPH were investigated. SPH displayed DPPH radical scavenging (IC 50  = 4.22 mg/mL), ABTS + radical scavenging (IC 50  = 2.93 mg/mL), reducing power and metal ion-chelating activities (IC 50  = 0.67 mg/mL). Furthermore, SPH significantly (P < .05) inhibited the generation of intracellular reactive oxygen species (ROS) in Caco-2 cells. After simulated GI digestion, the antioxidant properties of SPH were enhanced, except for a decrease in ABTS + radical scavenging activity. After transepithelial transport, the permeates maintained partial antioxidant activity and the LC-MS/MS data further identified the absorbed soybean peptides. These results suggest that SPH contains the antioxidant peptides that are potentially bioavailable and can be regarded as a promising source of functional food ingredients. [ABSTRACT FROM AUTHOR]

Abstract: BACKGROUND: To exert an antihypertensive effect after oral administration, angiotensin I‐converting enzyme (ACE)‐inhibitory peptides must remain active after intestinal transport. The purpose of this article is to elucidate the transport permeability and route of ACE‐inhibitory peptide Arg‐Leu‐Ser‐Phe‐Asn‐Pro (RLSFNP) across the intestinal epithelium using Caco‐2 cell monolayers. RESULTS: Intact RLSFNP and RLSFNP breakdown fragments F, FNP, SFNP and RLSF were found in RLSFNP transport solution across Caco‐2 cell monolayers using ultra‐performance liquid chromatography–tandem mass spectrometry. RLSFNP fragments FNP, SFNP and RLSF also contributed to ACE inhibitory effects. Protease inhibitors (bacitracin and leupeptin) and absorption enhancers (sodium glycocholate hydrate, sodium deoxycholate and Na2EDTA) improved the transport flux of RLSFNP. A transport inhibitor experiment showed that intact RLSFNP may be transported via the paracellular route. CONCLUSION: Intact RLSFNP can be transported across the Caco‐2 cell monolayers via the paracellular route. Extensive hydrolysis was the chief reason for the low permeability of RLSFNP. © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]