Selective depletion and measurement of glutathione in periportal and pericentral regions in the perfused rat liver.
A method was developed to selectively deplete and measure glutathione (GSH) in periportal and pericentral regions of the liver lobule based on the formation of chlorodinitrobenzene-glutathione (CDNB-GSH) adducts. Using microlight guides, we demonstrated that small pulses of CDNB caused reflected light at 366 nm to decline only in upstream regions of the liver lobule. This indicates that GSH was only depleted in upstream periportal or pericentral regions following perfusions in the anterograde or retrograde direction, respectively. Infusion of repeated pulses of CDNB in alternating directions of perfusion (up to eight) allowed selective and complete depletion of GSH in specific sublobular regions. Summation of local rates of adduct formation indicated that GSH content averaged 3.6 +/- 0.8 mumol/g in periportal regions and was 3.3 +/- 0.8 mumol/g in pericentral areas. Total values for GSH calculated from GSH-CDNB adduct formation were nearly identical to levels of GSH measured chemically. This new method may be useful in evaluating the mechanism of toxic chemicals which interact with GSH in discrete regions of the liver lobule.
Schön
M R
MR
Department of Pharmacology, University of North Carolina, Chapel Hill 27514.
Kauffman
F C
FC
Thurman
R G
RG
D000818:Animals; D004137:Dinitrochlorobenzene / Q000008:administration & dosage; D005260:Female; D005978:Glutathione / Q000032:analysis / Q000172:deficiency*; D007262:Infusions, Intravenous; D008099:Liver / Q000032:analysis*; D010477:Perfusion; D011168:Portal System / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D013053:Spectrophotometry / Q000379:methods
D004137:Dinitrochlorobenzene; D005978:Glutathione