Increased renal expression of bilirubin glucuronide transporters in a rat model of obstructive jaundice.
Regulation of bilirubin glucuronide transporters during hyperbilirubinemia in hepatic and extrahepatic tissues is not completely clear. In the present study, we evaluated the regulation of the bilirubin glucuronide transporters, multidrug resistance-associated proteins (MRP)2 and 3, in rats with obstructive jaundice. Bile duct ligation (BDL) or sham operation was performed in Wistar rats. Liver and kidneys were removed 1, 3, and 5 days after BDL (n = 4, in each group). Serum and urine were collected to measure bilirubin levels just before animal killing. MRP2 And MRP3 mRNA expressions were determined by real-time RT-PCR. Protein expression of MRP2 and MRP3 was determined by Western blotting. Renal MRP2 function was evaluated by para-aminohippurate (PAH) clearance. The effect of conjugated bilirubin, unconjugated bilirubin, human bile, and sulfate-conjugated bile acid on MRP2 gene expression was also evaluated in renal and hepatocyte cell lines. Serum bilirubin and urinary bilirubin excretion increased significantly after BDL. In the liver, the mRNA expression of MRP2 decreased 59, 86, and 82%, and its protein expression decreased 25, 74, and 93% compared with sham-operated animals after 24, 72, and 120 h of BDL, respectively. In contrast, the liver expression of MRP3 mRNA increased 138, 2,137, and 3,295%, and its protein expression increased 560, 634, and 612% compared with sham-operated animals after 24, 72, and 120 h of BDL, respectively. On the other hand, in the kidneys, the mRNA expression of MRP2 increased 162, 73, and 21%, and its protein expression increased 387, 558, and 472% compared with sham-operated animals after 24, 72, and 120 h of BDL, respectively. PAH clearance was significantly increased after BDL. The mRNA expression of MRP2 increased in renal proximal tubular epithelial cells after treatment with conjugated bilirubin, sulfate-conjugated bile acid or human bile. Upregulation of MRP2 in the kidneys and MRP3 in the liver may be a compensatory mechanism to improve bilirubin clearance during obstructive jaundice.
Tanaka
Yuji
Y
Third Department of Internal Medicine, Mie University School of Medicine, 2 - 174 Edobashi, Tsu City, 514 - 8507, Japan.
Kobayashi
Yoshinao
Y
Gabazza
Esteban C
EC
Higuchi
Kunihiro
K
Kamisako
Toshinori
T
Kuroda
Makoto
M
Takeuchi
Keisuke
K
Iwasa
Motoh
M
Kaito
Masahiko
M
Adachi
Yukihiko
Y
D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D001646:Bile; D001652:Bile Ducts / Q000601:surgery; D001663:Bilirubin / Q000031:analogs & derivatives* / Q000097:blood / Q000378:metabolism* / Q000494:pharmacology / Q000652:urine; D015153:Blotting, Western; D002779:Cholestasis / Q000378:metabolism*; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007668:Kidney / Q000737:chemistry / Q000378:metabolism*; D008026:Ligation; D008099:Liver / Q000737:chemistry / Q000378:metabolism; D026901:Membrane Transport Proteins; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000032:analysis / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar; D010130:p-Aminohippuric Acid / Q000378:metabolism
D001663:Bilirubin; D026901:Membrane Transport Proteins; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D012333:RNA, Messenger; D010130:p-Aminohippuric Acid; C000715087:ABCC2 protein, human; C114462:multidrug resistance-associated protein 3; C016266:bilirubin glucuronate; C421109:multidrug resistance-associated protein 1