Metabolism of lipid peroxidation product, 4-hydroxynonenal (HNE) in rat erythrocytes: role of aldose reductase.
Lipid peroxidation represents a significant source of erythrocyte dysfunction and aging. Because the toxicity of lipid peroxidation appears to be in part due to aldehydic end products, we examined, in rat erythrocytes, the metabolism of 4-hydroxy-trans-2-nonenal (HNE), one of the most abundant and toxic lipid-derived aldehydes. Packed erythrocytes, 0.1 ml, completely metabolized 20 nmoles of HNE in 20 min. The glutathione conjugate of HNE and 4-hydroxynonanoic acid (HNA) represented 70 and 25% of the total metabolism, respectively. Approximately 70% of the metabolites were extruded to the medium. Upon electrospray ionization mass spectrometry, the glutathione conjugate resolved into two distinct species corresponding to glutathionyl HNE (GS-HNE) and glutathionyl 1,4-dihydroxynonene (GS-DHN). The concentration of GS-DHN formed was twice that of GS-HNE. Inhibition of aldose reductase by sorbinil and tolrestat led to a selective decrease in the formation of GS-DHN, although the extent of HNE glutathiolation was unaffected. Inhibitors of aldehyde or alcohol dehydrogenase, i.e., cyanamide and 4-methyl pyrazole, had no effect on the formation of HNA and GS-DHN, indicating that these enzymes are not significant participants in the erythrocyte HNE metabolism. Thus, oxidation to HNA, conjugation with glutathione, and further reduction of the conjugate by aldose reductase appear to be the major pathways of HNE metabolism in erythrocytes. These pathways may be critical determinants of erythrocyte toxicity due to lipid peroxidation-derived aldehydes.
Srivastava
S
S
Division of Cardiology, University of Louisville, Louisville, KY, USA. ssrivaast@utmb.edu
Dixit
B L
BL
Cai
J
J
Sharma
S
S
Hurst
H E
HE
Bhatnagar
A
A
Srivastava
S K
SK
D000449:Aldehyde Reductase / Q000097:blood*; D000447:Aldehydes / Q000097:blood; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004912:Erythrocytes / Q000201:enzymology / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D015227:Lipid Peroxidation; D051381:Rats; D018629:Spectrometry, Mass, Secondary Ion; D014316:Tritium
D000447:Aldehydes; D014316:Tritium; D000449:Aldehyde Reductase; C027576:4-hydroxy-2-nonenal