17α, 20α-Dihydroxy-4-pregnen-3-one (17α, 20α-P) のRadioimmunoassayによる測定法と正常婦人並びに正常妊婦の血中動態
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17α, 20α-dihydroxy-4-pregnen-3-one (17α, 20α-P) is reportedly a hyper-tensinogenic substance in a model of ACTH induced hypertension in sheep (Scoggins et al). This steroid has not previously been measured in Japan. We have developed a new radioimmunoassay (RIA) system for 17α, 20α-P, and measured this steroid in peripheral blood samples from non-pregnant and pregnant women. In this clinical investigation concentrations of 17α-hydroxyprogesterone (17α-OHP) and progesterone (P) were also measured in the same clinical samples. Specific anti-serum against 17α, 20α-P-3-carboxymethyloxime-BSA was generated in rabbits. Cross-reactions with 5-pregnen-313, 17α 20α-triol (17α, 20α-P5) and 17α-OHP were 42 and 2.4%, respectively. Other steroids showed cross-reactivity of less than 1%. As an internal standard dexamethasone (500ng/100μl) was added to the samples which were extracted twice with 6 volumes of dichloromethane. Separation of the plasma extract was performed by high performance liquid chromatography (HPLC) with the ODS column in the solvent system, acetonitrile : water = 55 : 45. Eluates of the 17α, 20α-P and 17α-OHP fractions were collected separately and the solvent was evaporated in vacuo. RIA was carried out on each extract with specific anti-sera. RIA of P was performed independent-ly using the anti-P-3-CMO-BSA antiserum; 0.1 ml of serum was extracted with n-hexane and subjected to RIA without chromatography. The coefficient of variation for intra- and inter-assay (n = 10) were 8.5 and 11.7% for 17α, 20α-P; 7.2 and 12.1% for 17-OHP and 10.5 and 14.0% for P, respectively. The recovery rates of 17α, 20α-P, 17α-OHP and P were 95 ± 1.5, 93.2 ± 3.5 and 97.5 ± 4.2%, respectively. With the use of this method, 17α, 20α-P was measured in the blood samples from the menstrual cycle and normal pregnancy. The mean serum 17α, 20α-P concentrations ± SD in the follicular, ovulatory and luteal phases, and in the first, second and third trimesters were; 76 ± 6.0,100 ± 31,101 ± 6.4,226 ± 80,151 ± 48 and 367 ± 99pg/ml, respectively. While the concentrations of 17α-OHP and P in the respective timings were; 0.92 ± 0.5 and 5.3 ± 1.2, 1.8 ± 0.3 and 13.5 ± 4.2, 1.6 ± 0.6 and 68.7 ± 28, and 5.2 ± 1.1 and 128 ± 46ng/ml. The normal women in the ovulatory and luteal phase showed slightly higher levels of 17α, 20α-P as compared to the value in the follicular phase. Comparing to the values of steroid concentrations in the luteal phase, all the steroids increased significantly during pregnancy. Concentrations of all steroids in the third trimester were significantly elevated as compared to those in the first trimester of pregnancy. 17α, 20α-P and 17α-OHP showed a similar tendency to decrease in the second trimester and they increased again in the third trimester. The source of 17α, 20α-P in human is not entirely clear yet, but it may be postulated that this steroid is produced in the ovary and the fetoplacental unit.
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関連論文
- Pregnancy induced hypertension (PIH) における17α,20α-dihydroxy-4-pregnen-3-one(17α,20α-P) の意義とその産生源に関する研究
- 17α, 20α-Dihydroxy-4-pregnen-3-one (17α, 20α-P) のRadioimmunoassayによる測定法と正常婦人並びに正常妊婦の血中動態