Enhanced lipid peroxidation in tourniquet-release mice.
BACKGROUND
The pathogenesis of ischemia-reperfusion involves generation of reactive oxygen and resulting lipid peroxidation. However, investigation that ischemia-reperfusion following tourniquet release enhances lipid peroxidation is insufficient.


METHODS
Tourniquet was applied to a unilateral hind limb of mice for 3h followed by 5-, 15-, 30- and 60-min release. To examine superoxide production immunohistochemically in ischemia-reperfusion muscles, a primary antibody directed to 4-hydroxy-nonenal (HNE) was used. Furthermore, we analyzed 7alpha- and 7beta-hydroperoxycholest-5-en-3beta-ol, 7alpha- and 7beta-hydroxycholesterol, and 7-ketocholesterol by HPLC in the gastrocnemius muscles, kidneys, liver, heart and lungs of mice after 1-h reperfusion.


RESULTS
Increased HNE immunoreactivitiy was observed in the tourniquet-applied side of gastrocnemius muscles of hind limb particularly after 5-min reperfusion. All the oxysterols were significantly higher in the gastrocnemius muscles of the tourniquet-applied side than of the contralateral muscles. Oxysterols were elevated in the kidneys and the liver. Together with the presence of high blood urea nitrogen, these data indicate that the kidney is vulnerable to ischemia-reperfusion.


CONCLUSIONS
The enhanced oxidative stress due to ischemia-reperfusion appears to increase HNE in muscle and oxysterols by peroxidation not only in the gastrocnemius muscles but also in the kidneys and liver.
Adachi
Junko
J
Department of Legal Medicine, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Japan. adachi@med.kobe-u.ac.jp <adachi@med.kobe-u.ac.jp>
Kurisaki
Emiko
E
Kudo
Risa
R
Nakagawa
Kanako
K
Hatake
Katsuhiko
K
Hiraiwa
Kouichi
K
Ueno
Yasuhiro
Y
D000447:Aldehydes / Q000378:metabolism*; D000818:Animals; D002784:Cholesterol / Q000031:analogs & derivatives / Q000032:analysis / Q000378:metabolism; D006321:Heart / Q000502:physiology; D006614:Hindlimb / Q000098:blood supply / Q000378:metabolism; D007511:Ischemia / Q000378:metabolism*; D007668:Kidney / Q000378:metabolism; D015227:Lipid Peroxidation; D008099:Liver / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009132:Muscles / Q000098:blood supply / Q000378:metabolism*; D015427:Reperfusion Injury / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism; D013997:Time Factors; D014111:Tourniquets / Q000009:adverse effects* / Q000662:veterinary
D000447:Aldehydes; D013481:Superoxides; D002784:Cholesterol; C027576:4-hydroxy-2-nonenal