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산화적 스트레스에 의한 N'-methyl-N'-nitroguanidine의 유전독성증가
강진석(Jin Seok Kang),정기경(Ki Kyung Jung),서수경(Soo Kyung Suh),김주환(Joo Hwan Kim),이화옥(Hwa Ok Lee),정해관(Hai Kwan Jung),김승희(Seung Hee Kim),박순희(Sue Nie Park) 환경독성보건학회 2007 환경독성보건학회지 Vol.22 No.4
To investigate the possible enhancement of genotoxicity in stress environment, we examined the effect of genotoxic material in oxidative stress-induced condition using human cell line. Human lymphoblast cell line, TK6 was treated with hydrogen peroxide (H₂O₂) for induction of oxidative stress, and treated with N’-methyl-N’-nitroguanidine (MNNG), as a genetoxic material. We carried out MTS assay to set treatment doses. TK6 was treated with H₂O₂ as 6.75 (low dose) or 13.5 μM (high dose) for 2 h, and treated with MNNG as 0.117 (low dose), 0.234 (middle dose), 0.468 μM (high dose) for 2 h. As results, a treatment of MNNG induced DNA damage as dose dependently. And TK6 treated with H₂O₂ at low as well as high dose followed by MNNG treatment showed higher DNA damage compared to MNNG alone treated groups. Malondialdehyde, as a marker of lipid peroxidation was increased in H₂O₂ and MNNG treated groups. Real-time RT-PCR analyses for expression sion of several antioxidative enzymes showed that catalase mRNA and glutathione peroxidase 1 mRNA expression were decreased in H₂O₂ and MNNG treated groups. Taken together, we conclude that genotoxicity induced by MNNG is enhanced in a condition of oxidative stress induced by H₂O₂ and it suggests that it should be associated with induction of lipid peroxidation and decrease of antioxidant enzymes.
白鼠에서 疼痛에 미치는 Phenobarbital 效果의 再評價
소병겸,이화옥,김기원,조규박,은홍배 전북대학교 의과학연구소 1986 全北醫大論文集 Vol.10 No.4
Clinically, subhypnotic doses of barbiturates have been known to elicit hyperlgesia. In this experiment, effect of acute or chronic phenobarbital on the response to pain in rats was reevaluated by hot-plate method. To elucidate its mechanism, changes in β-endorphin contents and [^3H]-morphine binding of the rat midbrain as well as functional opiate receptor in vas deferens were also measured. Intraperitoneal injection of subanesthetic dose of phenobarbital induced initial hyperalgesia followed by successive analgesia, while chronic phenobarbital-treatment decreased reactivity to pain. Naloxone(10mg/kg, I. p.) markedly shortened hot plate latency, and significantly inhibited the analgesic action of phenobarbital. Single dose of phenobarbital did not affect β-endorphin contents and [^3H]-morphine binding in rat madbrain, but in the chronic phenobarbital-treated groups, β-endorphin content was increased, while Bmax of opiate receptor binding was decreased. Moreover, very significant correlations among response to pain, changes in β-endorphin contents and opiate receptor binding were observed. However, Kd values of opiate receptor binding swere not changed in all preparations. In the chronic phenobarbital-treated vas deferens preparation, ID_50 of morphine was increased with concomitant decrease of maximum effect. But pA_2 value for naloxone was not changed. From these resuts, it is suggested that phenobarbital can produce analgesia due to changes in β-endorphin contents as well as functional opiate receptors by receptor regulation.
白鼠의 腦內 Opiate 受容體의 日中 變動에 미치는 Phenobarbital의 影響
박영걸,김기원,조규박,정성후,이화옥 전북대학교 의과학연구소 1987 全北醫大論文集 Vol.11 No.1
실험적으로 명암을 조절한 백서 뇌내 specific opiate binding β-endorphin 함량을 시간별로 정량하여 그 일중변동 유무및 이에 대한 지속적인 암적응및 phenobarbital 장기처리에 의한 영향과 opiate receptor binding과 β-endorphin 함량 양자간의 관계를 관찰하여 다음과 같은 성적을 얻었다. 1. L : D, 12 : 12 주기에 적응시킨 대조군에서 maximum[3H]-morphine binding과 뇌내 β-endorphin 함량은 22시및 06시에 최고에 달하는 매우 유의한 일중변동을 보였고 24시간 평균[3H]-morphine binding 및 뇌내 β-endorphin 함량은 각각 0.45±0.03 pmole/㎎ protein과 46.7±3.6 fmole/mg protein이었다. 2. D : D, 12 : 12 주기에 적응시킨 표본에서 maxi-mum [3H]-morphine binding과 뇌내 β-endorphin 함량은 대조군에서와 다른 02시및 14시에 최고에 달하는 시간별로 유의한 일중변동을 보였고 24시간 평균 maximum [3H]-morphine binding치는 0.36±0.03 pmole/mg protein으로 대조군에 비하여 유의한 감소를 보였고, 24시간 평균 뇌내 β-endorphin치는 35.9±3.1fmole/mg protein으로 대조군에 비하여 유의한 감소를 보였다. 3. Phenobarbital 처리 표본에서 maximum [3H]-morphine binding과 뇌내 β-endorphin 함량은 02시 및 14시에 최고, 그리고 14시 및 02시에 최저에 달하는 대조군과는 다른 일중변동을 보였으며 24시간 평균 maximum [3H]-morphine binding 치는 0.33±0.03 pmole/mg protein으로 대조군에 비하여 유의한 감소를 보였고, 24시간 뇌내 β-endorphin 함량은 60.0±5.8 fmole/mg protein으로 유의한 증가를 보였다. 4. 전실험군에서 opiate receptor binding의 Kd치는 변동되지 않았다. 5. 전 실험군에서 maximum[3H]-morphine binding은 뇌내 β-endorphin 함량과 유의한 역상관 관계를 나타내었다. 이상의 실험성적은 phenobarbital이 뇌내 β-endorphin 함량과 opiate 수용체의 숫적 변동을 일으켜 morphine의 약리적 작용을 변동시킬 수 있으며, 이와 더불어 opiate 수용체와 β-endorphin 함량 을중변동을 변화시킬 수 있음을 보여준다. To investigate the influence of penobarbital sodium on the action of morphine and the diurnal rhythms of both opiate receptor binding and β-endorphin contents, the amount of specifically bound [3H]-morphine and immunoreactive β-endorphin were measured in the midbrain of phenobarbital-treated rats at 4-hr intervals in a day. Rats were housed and adapted to a controlled cycle of either 12-hr light -12-hr dark or 24-hr constant dark. After 3 weeks of adaptation, 0.5㎖ of physiological saline or phenobarbital sodium(20㎎/㎏/day) were administered intraperitoneally twice a day for 2 weeks. Highly significant diurnal rhythms of opiate receptor binding and β-endorphin were present in rat midbrain. In the control group, the peack of maximum [3H]-morphine binding was observed at 22:00 O'clock, whereas the peak of β-endorphin contents was found at 06:00 O'clock. Even in the absence of time cues these diurnal rhythms persisted, but they were highly modified with respect to the wave forms as well differences in the timing of peak and nadir. In the phenobarbital-th\reated group, these diurnal rhythms were also modified in shape, phase and ampltude, as well as in timing of peak and nadir. In the phenobaribital-treated group, these diurnal rhythms were also modified in shape, phase and amplitude, as well as in timing of peak and nadir. In this group, 24-hr mean of opiate receptor bindindg was significantly decreased, while the 25-hr mean level of β-endorphin content was highly increased. However, Kd values in all experimental groups was not changed. This indicates that dfferences in binding were not due to changes in the affinity but in the number of binding sites. Statistical analysis of of regression line indicates that changes in receptor binding are closely correlated with the changes of β-endorphin content. These results suggest that phenobarbital may influence the action of morphin by changing the number of opiate receptors and that ine modification of diurnal rhythm of opiate receptor by the agent is possibly due to changes in β-endorphin content.