Testosterone-Estradiol Binding Globulin (TeBG) のbinding capacity測定法について

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概要

• 論文の詳細を見る

• Estradiol (E<SUB>2</SUB>) binding capacity to plasma protein was estimated by simplified methods. Three techniques of dextran coated charcoal (D.C.), ammonium sulfate (A.S.) and concanavalin A sepharose (Con A.) were compared to separate bound and unbound hormones.<BR>The D.C. and A.S. methods proceeded as follows: Purified <SUP>3</SUP>H-E<SUB>2</SUB> in benzene-methanol was evaporated under nitrogen gas, and a half ml of sample plasma was added. Then, 0.4 ml of D.C. or A.S. was added to the plasma after one hour incubation at 37°C. Separation of protein bound E<SUB>2</SUB> was performed by centrifugation of 3000 r.p.m. at 4°C for 30 minutes. The supernate was counted by a liquid scintillation gamma counter.<BR>The following procedure was performed in the Con A. method : Sample plasma of 0.1 ml and Con A. of 0.3 ml were mixed for one hour. Then, <SUP>3</SUP> H-E<SUB>2</SUB> was added and mixed for 30 minutes and settled for two hours. The supernate and/or sediment were counted. This procedure could be performed at room temperature.<BR>Reproducible results were obtained when the prepared concentration of D.C. was 0.5%, A.S. was 50% and Con A. was 50%. The highest binding capacity was observed in alpha globulin fraction compared with beta and gamma globulin. The observed binding percents in alpha globulin were 33.5% by D.C., 78.1% by A.S. and 55.6% by the Con A. method.<BR>A significant decrease of binding percent was observed in alpha globulin fraction when the protein solution was preincubated with cold E<SUB>2</SUB>, suggesting the responsible role of alpha globulin in E<SUB>2</SUB> binding.<BR>Plasma obtained from normal men, and women with normal periods, pregnancy, first degree amenorrhea and second degree amenorrhea were examined by the three methods. The D.C. method showed a notable difference between these plasma samples. The obtained percent bound was 84.6% in pregnancy of late trimester, 74.5% in normal women, 60% in amenorrheas and 45% in normal men.<BR>These results suggest that alpha globulin might play an important role in E<SUB>2</SUB> binding, and the D.C. method could be suitable for the routine study of estradiol binding in women of different estradiol levels.

• 日本内分泌学会の論文

著者

• 宇津宮 隆史

  九州大学温泉治療学研究所産婦人科

• 門田 徹

  九州大学温泉治療学研究所産婦人科

• 片桐 英彦

  福岡赤十字病院産婦人科

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