| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2000: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1999: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Research Abstract |
We investigated whether administration of antisense oligonucleotide for interleukin-1 receptor antagonist (IL-1ra) mRNA into the lateral ventricle of rat brain could diminish concentrations of the cytokine in the brain, and whether stress-induced enhancement of the cytokine levels could be inhibited by administrating the antisense oligonucleotide. Arrangement of both antisense oligonucleotide and miss-matched one was determined by using ANTISENSER (Biognostik, Gottingen, Germany). Inhibitory effect of antisense oligonucleotide was estimated by NR8383 derived from rat alveolar macrophage. Two nano mole of oligonucleotide was injected into the left lateral ventricle of rats (Sprague-Dawley, male, 12-14 week old). The rats were decapitated under anesthesia with ether and pentobarbital at 12 h after the injection, and the brains were removed from the skull and homogenized with 10-times volume of culture medium on ice. Supernatant of the medium was collected after centrifuge (15,000 rpm, 10
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min) and stored at -80 * until measurement of IL-1ra levels. In the series of experiments to investigate effect of stress exposure, supernatant was extracted from rats that had been immobilized for 60 min. The experiments conform to the standards put forth in the Guideline for the Care and Use of Laboratory Animals of Keio University School of Medicine.
Adding antisense oligonucleotide into the culture medium significantly inhibited concentration of IL-1ra produced by NR8383. Administration of antisense oligonucleotide into the rat ventricle did not decrease in the concentration of IL-1ra in the brain, compared with that of miss-matched one. Stress exposure did not enhance the levels of IL-1ra in the rat brain, and administration of antisense oligonucleotide had no effect on the concentration of IL-1ra in the brain of the stress-exposed rats.
These results suggest that an alteration of timing for injection into the brain of animals was necessary to lead inhibitory effect of antisense oligonucleotide sufficiently. Less
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