| Project/Area Number |
14370178
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Gastroenterology
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| Research Institution | Mie University |
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Principal Investigator |
ADACHI Yukihiko Mie University, Faculty of Medicine, Professor, 医学部, 教授 (50111026)
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| Co-Investigator(Kenkyū-buntansha) |
KOBAYASHI Yoshinao Mie University, University Hospital, Research Associate, 医学部附属病院, 助手 (70378298)
玉置 繁憲 三重大学, 医学部附属病院, 助手 (80260602)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥12,000,000 (Direct Cost: ¥12,000,000)
Fiscal Year 2004: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2003: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2002: ¥7,100,000 (Direct Cost: ¥7,100,000)
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| Keywords | transporter / organic anion / polymorphism / Uridine-diphosphate glucuronosyl transferase / multidrug resistance-associated protein / organic anion transporting polypeptide / polypeptide / organic anion transporting polypeptide / Gunn rat / uridine-diphosphate glucuronosyltransferase / stem cell / bilirubin UDP-glucronosyltransferase / Eisai hyperbilirubinuria / short interfering RNA / 臓器相関 |
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| Research Abstract |
Various mutations of bilirubin UDP-glucronosyltransferase gene (UGT1A1) have been reported in patients with familial unconjugated hyperbilirubinemia. To clarify the incidence of this gene mutation in the Japanese population, we investigated the presence of UGT1A1 mutation in a group of Japanese patients with Crigler Najjar syndrome(CNS) type 2 and Gilbert's syndrome(GS), as well as in healthy anicteric subjects. Polymorphisms in the coding region of UGT1A1 were commonly observed in Japanese patients with GS and in healthy subjects. The genetic basis of hyperbilirubinemia appears to be different between the Japanese population and Caucasians. In this study, we additionally identified a novel mutation, heterozygous P364L. The P364L UGT1A1 enzyme activity was 64.4% lower than the wild type enzyme activity.
Gunn rat is an animal model of CNS type I. UGT1A1 has been reported to be deficient in CNS type 1. We evaluated expressions of organic anion transporting polypeptide(oatp) 1, oatp2, mult
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idrug resistance-associated protein(mrp) 2, and mrp3 in the liver and kidney of Gunn rats. Decreased expression of oatpl and increased expression of mrp3 were observed in the liver and kidney of Gunn rats.
Eisai hyperbilirubinuria rat(EHBR) is an animal model of Dubin-Johnson syndrome that suffers from jaundice due to impaired biliary excretion of bilirubin glucuronides. In EHBR, deficiency of mrp2 causes defective biliary excretion of numerous organic anions. Hepatic and renal expression of mrp3 was significantly higher in EHBR than in SD rats. Hepatic and renal expression of mrp1 and mrp6 mRNA was not significantly different between EHBR and SD rats. Mrp1 and mrp6 proteins were hardly expressed in liver and kidney of both EHBR and SD rats. In contrast to mrp3, hepatic expression of oatp1 and oatp2 mRNA were lower in EHBR than in SD rats. Immunohistochemistry disclosed that hepatic and renal mrp3 protein was localized at the basolateral membrane.
We developed a protein expression vector (pDEST_<26>) which includes the full length of human MRP2 cDNA, which is encapsulated by the envelope protein of the hemoagglutinating virus of Japan (HJV). Gene transfection with pDEST_<26> was performed on EHBR. We disclosed that transcellular transport of conjugated bilirubin was recovered in the liver from EHBR after transfection with MRP2. Serum conjugated bilirubin level decreased to normal levels in EHBR (35.7 μmol/L to 6.4 μmol/L) after transfection. Mrp3 expression decreased while Oatp1 or Oatp2 expression increased in the liver from transfected EHBR but these expressions were not significantly different from control SD rats. Less
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