Furosine 測定法を用いた glycosylated plasma proteins 測定の基礎的研究並びにその臨床的意義
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Glycosylated hemoglobin (Hb) has been clinically used as an indicator of long-term glucose control of diabetic patients. The main attachment site of glucose to Hb appears to be both ε-amino group of lysine and amino terminal valine residues. Fructose-lysine, which is formed by binding of glucose to ε-amino group of lysine residue, is changed into furosine [e-N-(2-furoylmethyl)-L-lysine] after acid hydrolysis. In the present study, furosine determination was used for the evaluation of glycosylated plasma proteins (GPP) in clinical practice. 1. Furosine was determined by high-performance liquid chromatography (HPLC) after acid hydrolys is of plasma protein. GPP was measured under the following conditions. 1) Amount of plasma protein : 50 mg 2) Duration of acid hydrolysis : 30 hours 3) Eluent: 7mM H_3PO_4 (pH 2.45) 4) Room temperature during operation by HPLC : 20-25℃ 5) UV detector wavelengths: 280 nm and 254 nm 6) The furosine content was expressed as the ratio of furosine peak area to tyrosine peak area in samples applied to HPLC. This method showed an excellent precision (intra-assay coefficent of variation was 1.4% and inter-assay coefficient of variation was 3.6%). 2. Stable component of GPP was specifically measured by the determination of furosine. 3. Glycosylation of plasma protein increased in proportion to the concentration of glucose and incubation time. 4. Positive correlation was found between furosine values and GPP levels determined by affinity chromatography (r=0.78, p<0.001). 5. The furosine values was 4.6±1.5% (mean±S.D.) in diabetic patients, being significantly higher than that found in normal subjects which was 2.6 ± 0.4%. The furosine values correlated with the fasting blood glucose level of two weeks before sampling. These results suggest that GPP can be useful as an indicator of blood glucose control of about two weeks before, and moreover, may be useful in the investigation of diabetic complications.
- 神戸大学の論文
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