Transport of cowpea bean derived peptides and their modulator effects on mRNA expression of cholesterol-related genes in Caco-2 and HepG2 cells

This work studied the cell transport of peptidase-generated peptides from cowpea bean proteins and their effects on mRNA expression of cholesterol-related genes in intestinal and liver cells. The <= 3 kDa hydrolysate was obtained and incubated with Caco-2 intestinal cells using Transwell center dot plates. HepG2 liver cells were incubated with synthetic analogues of peptides (MELNAVSVVHS and MELNAVSVVSH) identified by "de novo" peptide sequencing in the Caco-2 monolayer permeates. The mRNA expression of NPC1L1, ABCA1 and ABCG1 was measured in Caco-2 cells, in the presence or absence of <= 3 kDa hydrolysate and the expression of HMGCR, SREBP2, LXRa, AMPK1, was determined in the HepG2 cells in the presence or absence of synthetic peptides. Exposure of Caco-2 cells to cowpea <= 3 kDa hydrolysate (2.5 and 5 mg/mL) increased ABCG1 expression at 6 h and 12 h. SREBP2, HMGCR and LDLR mRNA levels were reduced in HepG2 cells after 24 h of treatment with MELNAVSVVHS peptide (50 mu M and 100 mu M). These results suggest that MELNAVSVVHS peptide is able to cross intestinal barrier and to modulate genes involved in cholesterol homeostasis.

Transport of cowpea bean derived peptides and their modulator effects on mRNA expression of cholesterol-related genes in Caco-2 and HepG2 cells

This work studied the cell transport of peptidase-generated peptides from cowpea bean proteins and their effects on mRNA expression of cholesterol-related genes in intestinal and liver cells. The <= 3 kDa hydrolysate was obtained and incubated with Caco-2 intestinal cells using Transwell center dot plates. HepG2 liver cells were incubated with synthetic analogues of peptides (MELNAVSVVHS and MELNAVSVVSH) identified by "de novo" peptide sequencing in the Caco-2 monolayer permeates. The mRNA expression of NPC1L1, ABCA1 and ABCG1 was measured in Caco-2 cells, in the presence or absence of <= 3 kDa hydrolysate and the expression of HMGCR, SREBP2, LXRa, AMPK1, was determined in the HepG2 cells in the presence or absence of synthetic peptides. Exposure of Caco-2 cells to cowpea <= 3 kDa hydrolysate (2.5 and 5 mg/mL) increased ABCG1 expression at 6 h and 12 h. SREBP2, HMGCR and LDLR mRNA levels were reduced in HepG2 cells after 24 h of treatment with MELNAVSVVHS peptide (50 mu M and 100 mu M). These results suggest that MELNAVSVVHS peptide is able to cross intestinal barrier and to modulate genes involved in cholesterol homeostasis.