RESUMO
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 ≤3â¯kDa hydrolysate was obtained and incubated with Caco-2 intestinal cells using Transwell® 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 ≤3â¯kDa hydrolysate and the expression of HMGCR, SREBP2, LXRα, AMPK1, was determined in the HepG2 cells in the presence or absence of synthetic peptides. Exposure of Caco-2 cells to cowpea ≤3â¯kDa hydrolysate (2.5 and 5â¯mg/mL) increased ABCG1 expression at 6â¯h and 12â¯h. SREBP2, HMGCR and LDLR mRNA levels were reduced in HepG2 cells after 24â¯h of treatment with MELNAVSVVHS peptide (50⯵M and 100⯵M). These results suggest that MELNAVSVVHS peptide is able to cross intestinal barrier and to modulate genes involved in cholesterol homeostasis.
Assuntos
Colesterol/metabolismo , Mucosa Intestinal/metabolismo , Metabolismo dos Lipídeos/genética , Fígado/metabolismo , Oligopeptídeos/metabolismo , Proteínas de Vegetais Comestíveis/metabolismo , RNA Mensageiro/metabolismo , Vigna/metabolismo , Células CACO-2 , Regulação Enzimológica da Expressão Gênica , Células Hep G2 , Homeostase , Humanos , Absorção Intestinal , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Oligopeptídeos/síntese química , RNA Mensageiro/genética , Sementes/metabolismoRESUMO
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.
RESUMO
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.
