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[磷酸吡哆醛在测定脑组织转氨酶活性中的应用]

[The use of pyridoxal-5-phosphate in determining aminotransferase activity in brain tissue].

作者信息

Rozanov V A, Karpovich G A, Muliukina N A

出版信息

Vopr Med Khim. 1987 May-Jun;33(3):129-32.

PMID:3630009
Abstract

When aspartate- and alanine transaminase (AST and ALT, respectively) activities were studied in homogenates of rat cerebellum, brain cortex and brain stem (using a modified procedure by Raitman and Frenkel), addition of 50 microM pyridoxal-5-phosphate (PALP) increased 2-3-fold the activity studied. With an increase in PALP concentration from 6.25 microM up to 200 microM AST- and ALT-activities increased dose-dependently, while at 100-200 microM concentration of PALP saturation of the reaction occurred and 200 microM of PALP decreased the AST activity. 10 min preincubation of these homogenates without and in presence of 0.25 mM PALP led to a distinct increase in amount of ketoderivatives reacting positively with 2,4-dinitrophenyl hydrazine, most markedly elevated in the sample containing PALP. At the same time, prolongation of the preincubation period under standard conditions in presence of an excess of the substrates and 50 microM PALP did not cause any increase in AST- and ALT-activities. During studies of the transaminase activity in brain biopsy, in the material obtained after neurosurgical operations as well as in medico-biological experiments, the activity should be estimated using two sets of conditions: under standard conditions and in presence of 50-100 microM PALP but without preincubation with PALP.

摘要

当在大鼠小脑、大脑皮层和脑干的匀浆中研究天冬氨酸转氨酶和丙氨酸转氨酶(分别为AST和ALT)活性时(采用Raitman和Frenkel改良的方法),添加50微摩尔磷酸吡哆醛(PALP)可使所研究的活性提高2 - 3倍。随着PALP浓度从6.25微摩尔增加到200微摩尔,AST和ALT活性呈剂量依赖性增加,而在100 - 200微摩尔PALP浓度时反应达到饱和,且200微摩尔PALP会降低AST活性。这些匀浆在无0.25毫摩尔PALP和有0.25毫摩尔PALP存在的情况下预孵育10分钟,导致与2,4 - 二硝基苯肼呈阳性反应的酮衍生物量明显增加,在含有PALP的样品中增加最为显著。同时,在存在过量底物和50微摩尔PALP的标准条件下延长预孵育时间,AST和ALT活性并未增加。在脑活检、神经外科手术后获得的材料以及医学生物学实验中研究转氨酶活性时,应使用两组条件来评估活性:在标准条件下以及在存在50 - 100微摩尔PALP但不与PALP预孵育的情况下。

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