Docosahexaenoic acid and eicosapentaenoic acid-enriched phosphatidylcholine liposomes enhance the permeability, transportation and uptake of phospholipids in Caco-2 cells.
The influence of docosahexaenoic acid (DHA)- and eicosapentaenoic acid (EPA)-enriched phosphatidylcholine (PC) on the permeability, transport and uptake of phospholipids was evaluated in Caco-2 cells. The cells were grown on permeable polycarbonate transwell filters, thus allowing separate access to the apical and basolateral chambers. The monolayers of the cells were used to measure lucifer yellow permeability and transepithelial electrical resistance (TEER). Transcellular transportation of diphenylhexatriene (DPH) labeled-PC small unilamellar vesicles (SUV) from the apical to basolateral chamber, and uptake of the same SUV was monitored in the cell monolayers. Cell-membrane perturbation was evaluated to measure the release of lactate dehydrogenase and to determine the cell viability with sodium 2-(4-iodophenyl)-3-(4-nitrophenyl) -5-(2, 4-disulfophenyl)-2H-tetrazolium dye reduction assay. The lucifer yellow flux was 1.0 and 1.5 nmol/h/cm(2) with 50 microM PC, and 17.0 and 23.0 nmol/h/cm(2) with 100 microM PC when monolayers of Caco-2 cells were treated with DHA- and EPA-enriched PC, respectively. TEER decreased to 24 and 27% with 50 and 100 microM DHA-enriched PC, and to 25 and 30% with 50 and 100 microM EPA-enriched PC, respectively. Our results show that DHA- and EPA-enriched PC increases tight junction permeability across the Caco-2 cell monolayer whereas soy PC has no effect on tight junction permeability. Transportation and uptake of DHA- and EPA-enriched PC SUV differed significantly (P < 0.01) from those of soy PC SUV at all doses. We found that PC SUV transported across Caco-2 monolayer and was taken up by Caco-2 cells with very slight injury of the cell membrane up to 100 microM PC. Lactate dehydrogenase release and cell viability did not differ significantly between the treatment and control, emphasizing that injury was minimal. Our results suggest that DHA- and EPA-enriched PC enhance the permeability, transport and uptake of PC SUV across monolayers of Caco-2 cells.
Hossain
Zakir
Z
Division of Marine Biosciences, Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, 041-8611, Japan. zakirh1000@yahoo.com
Kurihara
Hideyuki
H
Hosokawa
Masashi
M
Takahashi
Koretaro
K
D001692:Biological Transport / Q000187:drug effects*; D018938:Caco-2 Cells / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004281:Docosahexaenoic Acids / Q000494:pharmacology*; D015118:Eicosapentaenoic Acid / Q000494:pharmacology*; D017097:Electric Impedance; D005227:Fatty Acids / Q000737:chemistry; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008081:Liposomes / Q000494:pharmacology*; D010316:Particle Size; D010539:Permeability / Q000187:drug effects; D010713:Phosphatidylcholines / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism; D013024:Soybean Oil / Q000494:pharmacology; D019108:Tight Junctions / Q000187:drug effects; D022161:Transport Vesicles / Q000378:metabolism
D005227:Fatty Acids; D005456:Fluorescent Dyes; D008081:Liposomes; D010713:Phosphatidylcholines; D010743:Phospholipids; D004281:Docosahexaenoic Acids; D013024:Soybean Oil; D015118:Eicosapentaenoic Acid; D007770:L-Lactate Dehydrogenase