Transport of Antihypertensive Peptide RVPSL, Ovotransferrin 328-332, in Human Intestinal Caco-2 Cell Monolayers

J Agric Food Chem. 2015 Sep 23;63(37):8143-50. doi: 10.1021/acs.jafc.5b01824. Epub 2015 Sep 10.

Abstract

The objective of this study was to investigate the transepithelial transport of RVPSL (Arg-Val-Pro-Ser-Leu), an egg-white-derived peptide with angiotensin I-converting enzyme (ACE) inhibitory and antihypertensive activity, in human intestinal Caco-2 cell monolayers. Results revealed that RVPSL could be passively transported across Caco-2 cell monolayers. However, during the process of transport, 36.31% ± 1.22% of the initial RVPSL added to the apical side was degraded, but this degradation decreased to 23.49% ± 0.68% when the Caco-2 cell monolayers were preincubated with diprotin A (P < 0.001), suggesting that RVPSL had a low resistance to various brush border membrane peptidases. When transport from the apical side to the basolateral side was investigated, the apparent permeability coefficient (Papp) was (6.97 ± 1.11) × 10(-6) cm/s. The transport route of RVPSL appears to be the paracellular pathway via tight junctions, as only cytochalasin D, a disruptor of tight junctions (TJs), significantly increased the transport rate (P < 0.001). In addition, the relationship between the structure of RVPSL and transport across Caco-2 cell monolayers was studied by mutation of RVPSL. It was found that N-terminal Pro residues were more beneficial for transport of pentapeptides across Caco-2 cell monolayers than Arg and Val. Furthermore, RVPSL could be more easily transported as smaller peptides, especially in the form of dipeptides and tripeptides.

Keywords: Caco-2 cell monolayer; antihypertensive peptide; brush border membrane peptidase; egg white; transport.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Angiotensin-Converting Enzyme Inhibitors
  • Antihypertensive Agents*
  • Biological Transport
  • Caco-2 Cells
  • Chromatography, High Pressure Liquid
  • Egg Proteins / chemistry
  • Egg Proteins / metabolism
  • Egg Proteins / pharmacokinetics*
  • Epithelium / metabolism
  • Humans
  • Intestinal Mucosa / metabolism*
  • Microvilli / enzymology
  • Peptide Fragments / chemistry
  • Peptide Fragments / metabolism
  • Peptide Fragments / pharmacokinetics*
  • Peptide Hydrolases / metabolism
  • Structure-Activity Relationship

Substances

  • Angiotensin-Converting Enzyme Inhibitors
  • Antihypertensive Agents
  • Egg Proteins
  • Peptide Fragments
  • arginyl-valyl-prolyl-seryl-leucine
  • Peptide Hydrolases