Fumonisin B1 alters sphingolipid metabolism and disrupts the barrier function of endothelial cells in culture.
Fumonisins are responsible for at least two diseases of veterinary importance (equine leukoencephalomalacia and porcine pulmonary edema) and are carcinogenic for experimental animals and, perhaps, humans. They have been found to disrupt sphingolipid metabolism in many types of cells, including hepatocytes, neurons, and renal cells. In this study, endothelial cells form porcine pulmonary arteries were cultured on micropore filters as a model for the endothelial barrier, and barrier function was quantitated as the movement of albumin across the endothelial monolayers. Fumonisin B1 increased the amount of free sphinganine by 20- to 30-fold within 3 hr, as expected for inhibition of sphinganine (sphingosine) N-acyltransferase by this mycotoxin. At 30 to 50 microM, fumonisin B1 doubled the rate of albumin transfer across endothelial monolayers; however, there was no loss of cell viability based on morphology or trypan blue exclusion. When 15 microM D-erythro-sphinganine was added to the cells, the rate of albumin transfer also doubled (after 24 hr incubation) without a loss of viability; however, this treatment increased the cellular level of sphinganine by > 100-fold. Addition of 25 microM sphinganine caused even greater albumin transfer, but also resulted in significant cell death. These results establish that fumonisin B1 and D-erythro-sphinganine allow accelerated passage of macromolecules across the endothelium. Fumonisin B1 alters sphingolipid biosynthesis with an elevation of sphinganine in the cells which may, at least in part, explain the observed disruption of endothelial barrier function.
Ramasamy
S
S
Department of Nutrition and Food Science, University of Kentucky, Lexington, USA.
Wang
E
E
Hennig
B
B
Merrill
A H
AH
D000418:Albumins / Q000378:metabolism; D000704:Analysis of Variance; D000818:Animals; D002450:Cell Communication / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000032:analysis / Q000378:metabolism*; D037341:Fumonisins; D009183:Mycotoxins / Q000633:toxicity*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D011651:Pulmonary Artery; D015398:Signal Transduction / Q000187:drug effects; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism*; D013552:Swine; D014343:Trypan Blue / Q000378:metabolism
D000418:Albumins; D004791:Enzyme Inhibitors; D037341:Fumonisins; D009183:Mycotoxins; C056933:fumonisin B1; D011493:Protein Kinase C; D014343:Trypan Blue; D013110:Sphingosine; C005682:safingol