Protective effects of bilirubin against cyclophosphamide induced hemorrhagic cystitis in rats.
OBJECTIVE
The end product of the heme oxygenase pathway, bilirubin, is the most abundant endogenous antioxidant in mammals. We report the heme oxygenase-1 mediated production of bilirubin and its cytoprotective roles in cyclophosphamide induced hemorrhagic cystitis in rats.


METHODS
Female Sprague-Dawley rats received intraperitoneal administration of cyclophosphamide. In the first experiment hemin (an inducer of heme oxygenase-1) with or without zinc protoporphyrin IX (an inhibitor of heme oxygenase activity) was given before cyclophosphamide injection. Endogenous bilirubin production was analyzed in bladder tissues immunohistochemically. In another experiment bilirubin solution was administered before the cyclophosphamide injection. Changes in bladder weight, microscopic feature and expression levels of inducible nitric oxide synthase, proinflammatory cytokines and heme oxygenase were evaluated using polymerase chain reaction and immunostaining.


RESULTS
Bilirubin was generated in bladders with cyclophosphamide induced cystitis, especially in the urothelium and suburothelium. Hemin pretreatment provided increased production of endogenous bilirubin, which was decreased by zinc protoporphyrin IX. In an evaluation of the roles of bilirubin exogenous bilirubin administration ameliorated cyclophosphamide induced inflammatory changes and reduced the increase in bladder weight. The elevated expression of inducible nitric oxide synthase and interleukin-1beta in cyclophosphamide induced cystitis was significantly down-regulated by exogenously applied bilirubin. The expression of heme oxygenase-1 and 2 was not modified by bilirubin administration.


CONCLUSIONS
Cyclophosphamide induced hemorrhagic cystitis is accompanied by endogenous bilirubin production through heme oxygenase-1 induction in the bladder. Bilirubin has cytoprotective roles in association with the down-regulation of inducible nitric oxide synthase expression. Our results suggest that bilirubin may have therapeutic potential against bladder inflammatory insults such as cyclophosphamide induced cystitis.
Matsuoka
Yoh
Y
Department of Urology, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan. yoh-m.uro@tmd.ac.jp
Masuda
Hitoshi
H
Yokoyama
Minato
M
Kihara
Kazunori
K
D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D001663:Bilirubin / Q000096:biosynthesis*; D003520:Cyclophosphamide / Q000009:adverse effects; D003556:Cystitis / Q000139:chemically induced / Q000378:metabolism / Q000517:prevention & control*; D005260:Female; D051547:Heme Oxygenase-1 / Q000096:biosynthesis; D006470:Hemorrhage; D009676:Noxae / Q000009:adverse effects; D051381:Rats; D017207:Rats, Sprague-Dawley
D000975:Antioxidants; D009676:Noxae; D003520:Cyclophosphamide; D051547:Heme Oxygenase-1; D001663:Bilirubin