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Brain and Nerve No to Shinkei Volume 37, Issue 6 （June 1985）

Brain and Nerve 脳と神経 37巻6号（1985年6月発行）

Japanese English

原著

虚血脳におけるmannitol, vitamin E,betamethasoneおよび三剤併用時の脳保護作用—ESR spin trapping法による検討 PROTECTIVE EFFECT OF RADICAL SCAVENGERS ON CEREBRAL INFARCTION:EXPERIMENTAL STUDY UTILIZING SPIN TRAPPING METHOD OF ESR 富永悌二 ¹ , 今泉茂樹 ¹ , 吉本高志 ¹ , 鈴木二郎 ¹ , 藤田雄三 ² Teiji Tomonaga ¹ , Shigeki Imaizumi ¹ , Takashi Yoshimoto ¹ , Jiro Suzuki ¹ , Yuzo Fujita ² ¹東北大学脳研脳神経外科 ²倉敷中央病院脳神経外科 ¹Division of Neurosurgery, Institute of Brain Diseases, Tohoku University School of Medicine ²Depertment of Neurosurgy, Kurashiki Central Hospital pp.555-560

発行日 1985年6月1日 Published Date 1985/6/1

DOI https://doi.org/10.11477/mf.1406205523

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Abstract 文献概要
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　抄録　虚血脳におけるmannitol，vitamin E，betamethasoneの効果を検討する目的で，37℃で窒素下に保存したラット脳ホモジネートを用いPBNによるspin trappingを試みESR測定を行った。無処置群（n＝6）は，断頭後脳ホモジネートを作製し37℃で窒素下に保存しながら，保存前，保存後30分，60分，120分の各時点で脳ホモジネートを採取し，PBN，NADPH，Fe-EDTA添加による反応液を2つずつ作製した後，更返に20分閥で37℃それぞれ空気中あるいは窒素下にふ置してESR測定を行った。薬剤投与群は20％mannitol 10ml/kg（n＝6），vitamin E 30mg/kg（n＝6），betamethasone lmg/kg（n＝6）およびこれら三剤併用（n＝6）で，断頭30分前に静注し，同様の操作にてESR測定を行った。本実験で得られたシグナルは，無処置群および薬剤投与群で結合定数A＝16.0-16.6G，A＝3.0-3.8Gであった。そのシグナル強度は，無処置群で反応液の空気中ふ置の場合，窒素下保存時間に依存して増加し，窒素下ふ置の場合は，保存後60分にピークを認めた。薬剤投与群は，いずれの場合も強皮の減弱を認めたが，特に三剤併用投与群では著明な抑制効果が得られた。

　To evaluate the scavenging effect of mannitol, vitamin E and betamethasone in cerebral ischemia, spin trapping technique was applied to the detection of the free radicals generated in ischemic brain homogenate.

　Thirty Wistar rats were used for this study. In the control group, the brain homogenate prepared immediately after decapitation was preserved at 37℃ under N2 gas. Before the preservation and at 30 min, 60 min and 120 min from the start of the preservation, two reaction mixtures containing of spin trapping reagent phenylt-buthylnitrone (PBN), NADPH, Fe-EDTA and brain homogenate was prepared from each brain sample-one to be incubated for 20 min at 37℃in air and one to be incubated in nitrogen gas under similar condition. Then the free radical adducts of PBN were measured by electrone spin resonance (ESR).

　In pre-treated group, mannitol, vitamin E and betamethasone were administered intravenously 30 min prior to the decapitation and spin adducts of PBN were detected by same procedures as in control group.

　The ESR spectra, which hyperfine coupling constants were AN=16.0-16.6G and A〓=3.0-3.8 G, were obtained from the reaction mixtures in each group. Analyses of their relative intensity in control group revealed that the formation of free radical adducts of PBN was increased dependent on the preservation period under aerobic incubation, and increased gradually for 60 min of preservation time followed by a decrease under anaerobic incubation. And the signal intensities obtained at each time period of preservation were consistently stronger when the reaction mixture was incubated in air than when incubated in nitrogen gas. On the other hand, in pre-treated group the formation of spin adducts of PBN was suppressed when reaction mixtures were incubated under both air and nitrogen gas. Especially marked suppression was observed in combined administration of these drugs.