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      • KCI등재

        Caffeine이 지구성 운동 수행능력에 미치는 영향과 억제성 아미노산에 의하여 활성화되는 이온통로의 조절작용

        김은경,김영표,천병옥,이계영,김연정,임백빈,조영욱,김창주,김성수 대한스포츠의학회 1999 대한스포츠의학회지 Vol.17 No.1

        To investigate the effect and mechanism of caffeine on endurance exercise, two experiments were performed. First, to test caffeine effect on aerobic exercise, 200-300g Sprague-Dawley rats were used and three groups, control group, low caffeine injection group and high caffeine injection group, were divided. Blood smpling by heart puncture were done at rest, after 30 min treadmill exercise, and after maximal exercise. Blood glucose, free fatty acid concentration were detected and following results were obtained. Glucose concentration showed significant difference between groups(p=0.0305) and also significant changes were exhibited between time(p=0.0004). Free fatty acid concentration had no difference between groups. but had significance between times(p=0.00065). Exercise endurance performance time showed significant difference(p=0.02350 in high caffeine injection group compared to control group. In this experiments, endurance exercise capacity was increased by caffeine injection. Therefore, second experiment was performed to investigate the effect of caffeine on ion current induced inhibitory amino acid neurotransmitter. GABA and glycine. Single periaqueductal gray neuron was acutely dissociated and nystatin perforated patch clamp was performed under voltage clamping condition. Caffeine evoked outward current in PAG neuron dose dependent manner. 1mM of caffeine application had no response. but 3mM caffeine evoked about 32.5±8.539pA outward ion current and 10mM caffeine evoked about 215.46±19.4pA outward current. 10^-2mM GABA activated Cl ̄current and recorded by inward current. Caffeine inhibited GABA activated Cl ̄ current concentration dependent manner. 10^-2mM of caffeine had no effect on 1-^-2mM of GABA response. but 10^-1mM caffeine inhibited GABA activated Cl ̄ current about 5.74±2.13%, 1mM caffeine inhibited about 17.25±2.70%, 10mM caffeine inhibited GABA response about 45.31±7.71%. 10^-1mM of glycine activated Cl ̄ current and also recorded by inward current. Caffeine inhibited glycine activated Cl ̄ current concentration dependent manner. 10^-2mM caffeine decreased glycine activated Cl ̄ current about 4.61±1.650%, 10^-1mM caffeine decreased about 6.49±2.24%, 1mM caffeine decreased about 26.82±4.27%, and 10mM caffeine decreased glycine response about 94.47±1.39%. These results suggest that caffeine inhibite inhibitory amino acid, GABA and glycine, this response causes excitation of CNS and this seems to be the basic mechanism of increasing effect to aerobic exercise performance by caffeine.

      • KCI등재후보

        환경 오염물질의 진보된 독성 평가 기법

        류재천,최윤정,김연정,김형태,방형애,송윤선 한국환경독성학회 1999 환경독성보건학회지 Vol.14 No.1

        Recently, several new methods for the detection of genetic damages in vitro and in vivo based on molecular biological techniques were introduced according to the rapid progress in toxicology combined with cellular and molecular biology. Among these methods, mouse lymphoma thymidine kanase (tk) gene forward mutation assay, single cell gel electrophoresis (comet assay) and transgenic animal and cell line model as a target gene of lac I (Big Blue) and lac Z (Muta Mouse) gene mutation are newly introduced based on molecular toxicological approaches. The mouse lymphoma tk^(+/-) gene assay (MOLY) using L5178Y tk+i- mouse lymphoma cell line is one of the mammalian forward mutation assays, and has many advantages and more sensitive than hprt assay. The target gene of MOLY is a heterozygous tk^(+/-) gene located in 11 chromosome, so it is able to detect the wide range of genetic changes like point mutation, deletion, rearrangement, and mitotic recombination within tk gene or deletion of entire chromosome 11. The comet assay is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakages in mammalian cells. Also, transgenic animal and cell line models, which have exogenous DNA incorporated into their genome, carry recoverable shuttle vector containing reporter genes to assess endogenous effects or alteration in specific genes related to disease process, are powerful tools to study the mechanism of mutation in vivo and in vitro, respectively. Also in vivo acridine orange supravital staining micronucleus assay by using mouse peripheral reticulocytes was introduced as an alternative of bone marrow micronucleus assay. In this respect, there was an International workshop on genotoxicity procedure (IWGTP) supported by OECD and EMS (Environmental Mutagen Society) at Washington D, C, in March 25-26, 1999. The objective of IWGTP is to harmonize the testing procedures internationally, and to extend to finalization of OECD guideline, and to the agreement of new guidelines under the International Conference of Harmonization (ICH) for these methods mentioned above. Therefore, we introduce and review the principle, detailed procedure, and application of MOLY, comet assay, transgenic mutagenesis assay and supravital staining micronucleus assay.

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