Procyanidins can interact with Caco-2 cell membrane lipid rafts: involvement of cholesterol.

Large procyanidins (more than three subunits) are not absorbed at the gastrointestinal tract but could exert local effects through their interactions with membranes. We previously showed that hexameric procyanidins (Hex), although not entering cells, interact with membranes modulating cell signaling and fate. This paper investigated if Hex, as an example of large procyanidins, can selectively interact with lipid rafts which could in part explain its biological actions. This mechanism was studied in both synthetic membranes (liposomes) and Caco-2 cells. Hex promoted Caco-2 cell membrane rigidification and dehydration, effects that were abolished upon cholesterol depletion with methyl-ß-cyclodextrin (MCD). Hex prevented lipid raft structure disruption induced by cholesterol depletion/redistribution by MCD or sodium deoxycholate. Supporting the involvement of cholesterol-Hex bonding in Hex interaction with lipid rafts, the absence of cholesterol markedly decreased the capacity of Hex to prevent deoxycholate- and Triton X-100-mediated disruption of lipid raft-like liposomes. Stressing the functional relevance of this interaction, Hex mitigated lipid raft-associated activation of the extracellular signal-regulated kinases (ERK) 1/2. Results support the capacity of a large procyanidin (Hex) to interact with membrane lipid rafts mainly through Hex-cholesterol bondings. Procyanidin-lipid raft interactions can in part explain the capacity of large procyanidins to modulate cell physiology.

Bioavailability of intact proanthocyanidins in the rat colon after ingestion of grape seed extract.

Current evidence shows that monomeric flavonoids are known to be only slightly absorbed in the small intestine, but the metabolism of oligomeric and polymeric proanthocyanidins (PAC) in the colon is poorly understood. The objective of this study was to optimize the analysis of grape seed extract (GSE) in feces and use that method to assess the presence of PAC in the colon after ingestion of GSE. Rats were fed a diet ad libitum containing 0.25% (w/w) GSE for 10 days. Feces were collected daily and colonic contents at sacrifice on day 10, respectively. The recovery of fecal PAC using a solid-phase extraction (SPE) method was >70%. PAC were separated by normal-phase HPLC with fluorescence detection, and subsequent peak confirmation was done by MS-ion trap. The concentration of colonic contents at day 10 was 13.9 mg/kg for monomer, and those for oligomers (dimers-hexamers) were 33.4, 84.6, 87.2, 57.3, and 35.7 mg/kg, respectively. The concentration of monomeric and oligomeric PAC in daily feces was similar among days. In the mass balance analysis, approximately 11% of ingested PAC was recovered in the feces. These findings indicate that ingested PAC were present in the colon as the intact parent compounds and thus may contribute to the health of the gastrointestinal tract.

Large procyanidins prevent bile-acid-induced oxidant production and membrane-initiated ERK1/2, p38, and Akt activation in Caco-2 cells.

Procyanidins are oligomers of flavanol subunits present in large amounts in fruits and vegetables. Their consumption is associated with health benefits against colonic inflammation and colorectal cancer (CRC). Large procyanidins (with more than three subunits) are not absorbed by intestinal epithelial cells but could exert biological actions through their interactions with the cell membrane. This study investigated the capacity of hexameric procyanidins (Hex) to prevent oncogenic events initiated by deoxycholic acid (DCA), a secondary bile acid linked to the promotion of CRC. Hex interacted with Caco-2 cell membranes preferentially at the water-lipid interface. Hex (2.5-20 µM) inhibited DCA-triggered increase in cellular calcium, NADPH oxidase activation, and oxidant production. DCA promoted the activation of protein kinase B (Akt), of the mitogen-activated protein kinases ERK1/2 and p38, and of the downstream transcription factor AP-1. This activation was not triggered by calcium or oxidant increases. Hex caused a dose-dependent inhibition of DCA-mediated activation of all these signals. DCA also triggered alterations in the cell monolayer morphology and apoptotic cell death, events that were delayed by Hex. The capacity of large procyanidins to interact with the cell membrane and prevent those cell membrane-associated events can in part explain the beneficial effects of procyanidins on CRC.