Reduced Glutathione의 마우스 大腦組織內 移動에 關하여
Transport of Reduced Glutathione into the Brain Tissue of the Mouse
경북의대잡지 1973년 14권 2호 p.245 ~ 262
이한철, 朱永恩,
소속 상세정보
이한철 ( ) - 慶北大學校 醫科大學 生理學敎室
朱永恩 ( ) - 慶北大學校 醫科大學 生理學敎室
KMID : 0351619730140020245
Abstract
Reduced glutathione (GSH)의 組織細胞內로의 移動標相의一端을 알고져 마우스 大腦의 切片을 作成하고 이것을 1mM, 2mM 및 3mM의 GSH溶液中에 둔 후 4C,25C 및 37C의 溫度下에서 incubate하고 incubation中 5,15,30,60 및 120분에서 各各 組織內의 GSH 및 GSSG(dxidized glutahione)을 定量하고 이 兩者의 合計로서 總 SH基(sulfhydryl)의 量으로 하여 正常群 및 對照群의 그것과 比較한 結果 다음과 같은 結果를 얻었다.
1) 正常 마우스 大腦組織의 GSH, GSSG 및 총 SH基의 量은 各各 2.33±0.26, 1.72±0.27, 및 4.02±0.27mmol/gm wet.wt이었다.
2) 大腦組織의 GSH 및 GSSG의 量은 incubation의 온도가 4C alc 25C에서는 incubation溶液中의 GSH의 濃度와 無關하게 時間의 經過에 따라 큰 差異가 없었고 正常群 및 各對照群의 그것과 比較해서도 서로 비슷한 傾向을 나타내었다.
3) 37C에서 incubate 하였을 때의 大腦組織의 GSH,GSSG및 總 SH基의 量은 incubation溶液中의 GSH濃度에 比例하여 憎加하였고 또한 實驗時間 120分까지 繼續 增加되는 傾向을 나타내었다.
Glutathione, a tripeptide abundantly distributed among the animal and plant world, is known to exert some important physiological actions in the body, i. e. ; participation in various enzymatic reactions, membrane transport and action mechanism of some peptide hormones. Also, the effectiveness of reduced glutathione (GSH) for radioprotection has recently been established.
However, reports on the transport mechanism of GSH through the cell membrane is scanty, and little is known whether GSH is transported through the cell membrane by active transport or facilitated diffusion. or otherwise.
In the present study, an effort was made to observe the transport mechanism of GSH by using the brain tissue of the mouse as the model.
Whole brain was carefully removed from the normal mouse, and the brain slice of approximately 0.2mm in thickness was prepared manually.
The slice was incubated in the solution of GSH in the concentration of 1mM, 2mM and 3mM, and temperature of the incubation was set at 4C, 25C and 37C. At 5,15,30,60 and 120 min of the incubation, the slice was taken out and carefully washed with Krebs-Ringer phosphate buffer (KRP).
GSH level was measured by Ellman´s method, and GSSG (oxidized glutathione) was measured by the electrolytic reduction method described by Dohan and Woodward. The total SH (sufhydryl) group was calculated by the sum of GSH and GSSG. The control was set in each temperature group, in which the experimental procedures were identical as in the experimental groups but the incubation was carried out in the absence of GSH and onty in KRP.
Results thus obtained are summarized as follows.
1) Normal value of GSH, GSSG and total SH group of the mouse brain was 2.33±0.26, 1.72±0.27 and 4.03±0.27/㎛ol/gm wet wt., respectively.
2) when the incubation was carried out at 4C and 25C, GSH and GSSG levels of the mouse brain were similar in all the groups including the normal and control regardless of the concentration of GSH throughout the entire experiment.
3) when the incubation temperature was set at 37C, the levels of GSH, GSSG and total SH group increased in proportion to the concentration of GSH, and the increase was continuous till 120 min.
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