Localization of Xanthine Dehydrogenase in Liver

Hattori, Yukako; Nishino, Tokuzo; Usami, Chikako; Tsushima, Kenji; Ichikawa, Misao; Ichikawa, Atsushi

doi:10.1007/978-1-4684-5676-9_75

Cited by 2 publications

(1 citation statement)

References 14 publications

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“…Equal amounts of protein were resolved with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Mini-PROTEAN TGX 4–20%, Bio-Rad Laboratories, Inc., Hercules, California, USA), which was followed by electrophoretic transfer onto a polyvinylidene fluoride membrane with the iBlot system (Life Technologies Corporation, Grand Island, New York, USA). After blocking the nonspecific binding sites by incubation in 5% (w/v) skimmed-milk Tris-Buffered Saline-containing 0.1% Tween-20 (TBS-T) for 60 min, the membranes were incubated for 60 min at room temperature with 10,000-fold dilution of anti-XOR polyclonal rabbit antibody that was previously produced by Hattori et al 16) The membranes were further incubated with a horseradish peroxidase-conjugated anti-rabbit immunoglobulin antibody (Dako Denmark A/S, Glostrup, Denmark) for 60 min at room temperature at a 1:5,000 dilution. After washing in TBS-T, the membranes were reacted with ECL-prime (GE Healthcare, Uppsala, Sweden) at room temperature for 5 min.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of Neuronal Protective Effects of Xanthine Oxidoreductase Inhibitors on Severe Whole-brain Ischemia in Mouse Model and Analysis of Xanthine Oxidoreductase Activity in the Mouse Brain

Suzuki

Kusano

Matsuda

et al. 2015

Neurol. Med. Chir.(Tokyo)

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Global cerebral ischemia and reperfusion (I/R) often result in high mortality. Free radicals play an important role in global cerebral I/R. Xanthine oxidoreductase (XOR) inhibitors, such as allopurinol, have been reported to protect tissues from damage caused by reactive oxygen species (ROS) by inhibiting its production through XOR inhibition. The recently introduced XOR inhibitor febuxostat, which is a more potent inhibitor than allopurinol, is expected to decrease free radical production more effectively. Here, we analyzed the effects of allopurinol and febuxostat in decreasing global severe cerebral I/R damage in mice. Mice were divided into three groups: a placebo group, an allopurinol group, and a febuxostat group. Pathological examinations, which were performed in each group in the CA1 and CA2 regions of the hippocampus 4 days after I/R surgery, revealed that there was a decrease in the number of neuronal cells in the 14-min occlusion model in both regions and that drugs that were administered to prevent this damage were not effective. The enzymatic activity was extremely low in the mouse brain, and XOR could not be detected in the nonischemic and ischemic mice brains with western blot analyses. Thus, one of the reasons for the decreased effectiveness of XOR inhibitors in controlling severe whole-brain ischemia in a mouse model was the low levels of expression of XOR in the mouse brain.

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Section: Methodsmentioning

confidence: 99%