| Project/Area Number |
62570986
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Biological pharmacy
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| Research Institution | Okayama University |
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Principal Investigator |
OHMORI Shinji Okayama University, Professor, 薬学部, 教授 (10032872)
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| Co-Investigator(Kenkyū-buntansha) |
TSUBOI Seiji Okayama University, Assistant Faculty of Pharmaceutical Sciences, 薬学部, 助手 (50172052)
IKEDA Mikiko Okayama University, Lecturer, 薬学部, 講師 (20112154)
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| Project Period (FY) |
1987 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1989: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1988: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1987: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | D-Lactate / Methylglyoxal / L-Lactate / Pyruvate / High-performance liquid chromatography / After exercise / Diabetes mellitus / Starvation / 糖尿病ラット / 飢餓ラット / 2-ハイドロキシ6.7ジメトキシ-3-メチルキノキサリン / 脱水素酵素 / 蛍光分光定量 / 6,7ジメトキシ-3-メチル2-ハイドロキシキノキサリン / 運動 / D乳酸脱水素酵素 / HPLC / 2メチルキノキサノール / オルトフェニレンジアミン / 食事後 |
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| Research Abstract |
1. Establishment of the determination of D-lactate: D-Lactate was converted into pyruvate by D-lactate dehydrogenase. Hydrazine and a NADH-reoxidation-system were added to the reaction solution in order to complete the reaction. The formed pyruvate was converted into 2-hydroxy-3-methylquinoxaline (I) or 2-hydroxy-6,7-dimethoxy-3-methylquinoxaline (II), which were determined by HPLC with fluorescence detection. The determination limit was 600 fmol for (I) and 80 f mol for (II).
2. Formation of methylglyoxal (MG): L-Lactate was formed from MG. It was reported that MG was formed from several compounds in animals. These MG formation routes were reexaminated by our selective and sensitive determination method, because MG-determination methods hitherto used were not exact. As result, MG was found to be formed mainly from triosephosphates.
3. Changes in D-lactate and its related compound levels in blood plasma after exercise: Since MG was found to be correlated to energy and glucose metabolism by our experiments, D-lactate and its related metabolites in blood were determined. The D-lactate level was only about twice higher, not as much as L-lactate. The MG level, however, decreased in plasma, especially in red blood cells. D-Lactate and MG in: sweat and urine did not change after exercise.
4. Changes in D-lactate and its related compound content in blood and liver of diabetic and fasted rats: Phosphofructokinase activity of diabetic and fasted rats was not altered compared to a normal group, but pyruvate kinase activity decreased. The D-Lactate level in blood of diabetic rats increased to the double of the normal value, but MG and pyruvate decreased. The change of its L-lactate level was small. The D- and L-Lactate contents in liver of diabetic rats rised but the MG and pyruvate levels fell. In the case of starved rats, the D-lactate and MG levels in blood were elevated but pyruvate and L-lactate were decreased. D-lactate, MG and pyruvate levels fell.
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