Many works on actions of androgen to rat ventral prostate demonstarted that testoosterone increased RNA and protein syntheiss promptly after administration of testosterone. Before RNA synthesis was increased by testosterone, the fluctuation of ATP was noticed in ventral prostate. Increase of DNA syntheiss needed the lag time after testosterone in jection, and this increase was temporary.
Rat ventral prostate possessed 5a-reductase and 17-hydroxysteroid dehydrogenase of testosterone. The rate of 5a-reduction of testosterone was influenced by endogenous and exogenous testosterone level, but the rate of 17-dehydrogenation of testosterone was constant in various experimental conditions (Endocrinol. Jap. 16,453, 1969). The localization of 5a-reductase of testosterone was investigated by different centrifugation. Microsomal fraction contained about half of the content of RNA and more than half of the whole ATPase activity, however, the activity of 5a-reductase was appeared in some part. This results suggested some relationship between the presence of 5a-reductase and nuclear constituents.
The activity of 5a-reductase was found in human skin, and activity in scrotal skin was higher than that in abdominal part. This results suggested the importance of the role of 5a-reduction of testosterone in the hormonal action.

抄録全体を表示

Cyclic 3′, 5′-AMP has now been established as an intracellular second messenger mediating many of the actions of a variety of different hormones including ACTH. The level of this nucleotide depends upon the relative activities of at least two enzymes, viz., adenyl cyclase, which is at first activated by each hormone and catalyzes the conversion of ATP to cyclic AMP, and phosphodiesterase, which catalyzes the breakdown of cyclic AMP to 5′-AMP. It has been recently reported that the metabolic effects of catechol-amines are differentially affected by alpha and beta adrenergic agents. Stimulation of the alpha adrenergic receptor inhibits, and stimulation of the beta adrenergic receptor enhances the effects of catecholamines on cyclic AMP accumulation probably via their effects on adenyl cyclase. This is also the case with vasopressin and MSH. Few deta have been reported about the role of an adrenergic receptor mechanism in adrenal steroidogenic response to ACTH.
In the present study both alpha adrenergic blocker,, thymoxamine (5 to 50 μg/ml). and beta adrenergic stimulator, isoproterenol (1 μg/ml), were found to potentiate the rat adrenal steroidogenic response to ACTH in vitro, while both alpha adrenergic stimulator, methoxamine (1 to 20 μg/ml), and beta adrenergic blocker, alprenolol (10 to 25 μg/ml), to inhibit the response to ACTH. These results indicate that the general hypothesis (increase of hormone response by stimulation of the beta adrenergic receptor and decrease by stimulation of the alpha adrenergic receptor) may be valid in adrenal steroidogenic response to ACTH in vitro. Alpha adrenergic stimulator, phenylephrine (1 to 20μg/ml), and alpha adrenergic blocker, phentolamine (5 μg/ml), had no effect on the steroidogenesis induced by ACTH at the amounts used. Beta adrenergic blocker, propranolol (5 μg/ml), was shown to interefere with fluorescence of corticosterone by sulfuric acid-ethanol, resulting in the apparent potentiation of adrenal steroidogenesisby ACTH in vitro. Adenyl cyclase in rat adrenal homogenate was activated in the presence of isoproterenol and thymoxamine, and inhibited in the presence of propranolol, lending further support to the involvement of an adrenergic receptor mechanism in the adrenal. These adrenergic agents had no effect on phosphodiesterase activity. Prosta-glandin Ei (0.5 to 20μg/ml) was without effect on the steroidogenic response to ACTH. Omitting calcium from the medium and adding EGTA (1 mM) completely abolished the effect of ACTH on steroidogenesis and partially inhibited the dibutyryl cyclic-AMP-induced, steroidogenic response. Theophylline (1 mM) decreased rather than increased the steroidogenic action of ACTH and was without effect on the response to submaximal dose of dibutyryl cyclic AMP. On the contrary, imidazole (1 mM) potentiated the action of ACTH on steroidogenesis. These contradictory effects on adrenal steroidogenesis by ACTH may be explained by the possible action of these two agents on protein sythesis in rat adrenal tissue, a process which is essential for the steroidogenesis by ACTH. The fact that addition of cycloheximide (0.4 mg/ml) to medium inhibited completely the ACTH-induced steroidogenesis in vitro, also confirms the importance of new protein synthesis.

抄録全体を表示

TSH stimulates the secretion and synthesis of thyroid hormone, and also stimulates many metabolic pathways i.e, salt and water, carbohydrate, phosphorus and phospholipid, nucleic acid, amino acid and protein metabolism. Evidence has been accumulated that LATS causes a prolong effect on the thyroid and LATS differs in several respects from TSH. Until now the precise role of TSH and LATS at cellular level has not been known and the mode of action of two substances is not clear.
In this paper, the metabolic effects of TSH and LATS on the thyroid were examined extensively using both in vitro and in vivo systems. The incorporation of ³²P into the thyroid gland after a single injection of LATS and TSH was increased about 2 fold in the thyroxine treated mice. Also specific activities of RNA and phospholipid in the gland were increased remarkably. Experiments using ³H-uridine, TSH and LATS increased the incorporated radioactivity per RNA.
A single injection of TSH or LATS into mice falied to stimulate the incorporation of ³H-leucine into thyroidal protein. While the continuous injection of LATS for 5 days caused increased thyroid weight, augmented protein content and increased incorporation of ³H-leucine. Also the specific activity of incorporated radioactivity per mg thyroid protein was significantly increased. Treatment with LATS or TSH for 8 days produced marked goiter formation accompanied by a high I¹³¹ thyroidal uptake. LATS stimulates protein synthesis in the gland, as does TSH.
Using McKenzie's method, cyclic 3′5′ AMP (C 3′ 5′ AMP) and Dibutyryl cycilic 3′ 5′ AMP (D.C 3′ 5′ AMP) were found to increase thyroid hormone release from mouse thyroid in vivo. Other adenine nuclectides were also active. Simultaneous injection of theophylline and caffeine with C 3' 5' AMP and D.C 3′ 5′ AMP augmented the stimulatory effect of these adenine nucleotides on hormone release, although theophylline or caffeine had only a slight effect alone. No effect was obtained by the injection of NaF with D.C 3′ 5′ AMP. These adenine nucleotides have no enhancing action on hormone release from mouse thyroids during in vitro incubations, and no stimulatory effects were obtained by the nucleotides on the hydrolysis of thyroglobulin in mouse thyroid homogenates. These adenine nucleotides appeared to have no direct action on the enzyme participating in the breakdown of thyroglobulin.
According to Pastan's method, the effect of TSH on cyclase activity was determined using beef and rabbit thyroid homogenate, but the increase of cyclic AMP synthesis from ³H-ATP (or ¹⁴C-ATP) was not obtained by the large doses of TSH. Similarly also LATS haven't any stimulating action on adenyl cyclase activity. NaF did not elevate adenyl cyclase activity, but inhibited ATP destruction by the inhibition of ATPase. Theophylline inhibited cyclic AMP destruction significantly. Using beef thyroid slices, ³H-adenine was preincubated for 1 hr and then TSH (Armour or NIH) or LATS was incubated for 30 min. with theophylline or NaF. The incorporated radioactivity into cyclic AMP in whole thyroid slice was examined. TSH and LATS similarly stimulated the incorporation of ¹³¹H-adenine into cyclic AMP in beef thyroid slice, especially in the presence of theophylline. However, the stimulatory effect of NaF on cyclic AMP formation was very slight.
Both TSH and LATS elevated cyclic AMP levels in the gland, however it may be some of effects of both these thyroid stimulator. It is possible that TSH and LATS have similar stimulating action on various metabolism in the thyroid.

抄録全体を表示

In an attempt to elucidate the mode of hormone action in the most physiological situations, an in vivo-in vivo system (hormone was introduced in vivo, and the resulting biochemical responses were examined also in vivo) was studied by the use of an organ-microfluorometer. The instrument, developed by our hands for these particular ends, permits a faithful monitoring of alterations in the intracellular redox state of organs in situ, by continuously recording the fluorescence signal from reduced pyridine nucleotides which is compensated for the “hemodynamic artifact”. As examples of the study were presented the effect of thyrotropin and that of parathyroid hormone.
1. Thyrotropin (Thytropar, Armour) given iv to thyroxine-treated rabbits and rats caused a prompt and sustained fall in the level of reduced pyridine nucleotides in thyroid. Bovine serum albumin, growth hormone, prolactin, parathyroid hormone, luteinizing hormone, and follicle stimulating hormone did not reproduce the thyrotropin effect. Thyrotropin did not affect kidney, testis, or parathyroid pyridine nucleotides. Such oxidative shift of thyroid redox-sta te of pyridine nucleotides as produced by thyrotropin was mimicked by Dicoumarol or by long-acting thyroid stimulator. 3′, 5′-cyclic AMP or dibutyryl cyclic AMP, on the other hand, led to a reductive shift of the thyroid redoxstate. From these results a possibility is discussed that thyrotropin in vivo acts on thyroid to result, independently of the adenyl cyclase activation, in the oxidation of reduced pyridine nucleotides.
2. A parathyroid extract (Parathormone, Lilly) injected iv to rats induced a prompt and transient reduction in the kidney. This action was mimicked by calcium salts of poorly membrane-permeant anion like dextran-sulfate. Calcium salts of apparently permeant anion (e.g., aspartate) as well as EDTA or EGTA produced an oxidative response. The transient reduction was also reproduced by giving reductive substrates of the cytosol dehydrogenases. These results led to a hypothesis that parathyroid hormone causes a rise in the calcium ion activity in the cytosol compartment of renal tubules, resulting somehow in an accelerated production of the reductive substrate (s).

抄録全体を表示

The theory that insulin acts to facilitate the transmembrane transport of sugars possessing the same configuration as d-glucose about carbons 1, 2 and 3 has received substantial support from subsequent confirmatory observations, which have added another important point that insulin-sensitive sugar transfer occurs only in the membranes which restrict the entry of glucose into cells in the absence of insulin.
This fact has led us to assume that the facilitation of stereospecific sugar transfer should be caused by other peptide hormones than insulin in their target organs so long as the membranes of the organs confine sugars to a small fraction of intracellular water in the absence of the hormones concerned.
In this respect, characteristics of sugar transport were investigated in the bisected rabbit adrenal preparations, showing that the difference between water-H³ space and 1-arabinose-1-C¹⁴ space remains constant and 1-arabinose space is always in close proximity to sucrose space, i.e. the extracellular space. These findings indicate that the permeability barrier is operative in adrenal tissue. The addition of synthetici β^1-24 ACTH was shown to produce a significant increase in the d-xylose and 1-arabinose spaces in the adrenal preparations without causing any increase in the spaces of the corresponding optical isomers. Thus the proposal by Levine et al. concerning to the insulin action is compatible with the effect of synthetic ACTH on its target gland. The stereospecific effect on sugar transport was reproduced when cyclic AMP was added to the medium instead of synthetic ACTH. No effect of 5′-AMP was observed. Although prostaglandin E₁ diminished the response of sugar permeability of rabbit adrenals to synthetic ACTH, the effect of cyclic AMP on sugar transport was uninfluenced by the addition of prostaglandin E₁. It is quite possible from these data that β^1-24 ACTH alters the sugar transport system in adrenal cortex by stimulating the formation of cyclic AMP.
When bovine thyroid slices were incubated in the presence of a TSH preparation or cyclic AMP, there was also a stereospecific effect on sugar transport of the same type as observed in the rabbit adrenal preparations.
When blood or tissue concentrations are compared among hormones, cyclic AMP and main metabolic intermediates, it is tempting to suggest that cyclic AMP acts as an amplifier of hormone actions rather than a simple transmitter. Blood concentrations of almost all peptide hormones are 10^-10 molar range, while tissue concentrations of main metabolic intermediates such as glucose-6-P and ATP are 10^-3-10^-4 molar range. Cyclic AMP concentration in various tissues is just between the above two groups, varying with the absence or presence of hormones. The signal of hormones seems to be amplified markedly by fluctuations in cyclic AMP concentration in their target organs.
Since serum insulin concentration is 10^-10 molar range, it appears worthy to determine whether an amplifier may be involved in the mechanism of insulin action. The effects of various cyclic nucleotides on sugar transport in muscle preparations were examined. In intact rat diaphragms dibutyryl cyclic AMP did not increase the distribution space of 3-o-methylglucose. In frog sartorius muscles dibutyryl cyclic AMP and cyclic CMP caused a significant increase in 3-o-methylglucose space, but cyclic GMP, cyclic IMP and cyclic UMP had no such effect. It is noteworthy that there is an analogy in the chemical structure between adenine (6-amino purine) and cytosine (6-amino 2-oxypyrimidine).
Cyclic AMP appears not to be involved in the permeability response to insulin, but the possibility remains that the insulin action is mediated by another amplifier containing a 6-amino derivative of the purine or pyrimidine base.

抄録全体を表示

The mode of action of LH on testosterone biosynthesis in male rats and mice has been studied in vivo. In rat's experiments, testicular vein blood was collected and testosterone in the plasma was measured by fluorometry of Ohsawa et al (1969). After intravenous administration of LH testosterone output started to increase within three minutes and reached the maximum level in ten minutes. Thus, the effect of LH on testosterone secretion from rat's testis has been revealed to be quite rapid, like the effect of ACTH on adrenal cortex. The similar results have been obtained in mice. After intratesticular injection of LH, the content of testosterone in mouse testis started to increase within three minutes and reached the plateau in ten minutes.
Cyclic-3′, 5′ AMP, proposed mediator of action of LH in the Leidig cell, has been tested in the same assay system as described above, concerning the capacity of stimulation of testosterone synthesis in the male animals. Administration of cyclic AMP caused a prompt and significant increase in testosterone production in testes of both specieses. Cyclic AMP armarentiv mimics the action of LH.
Inhibitors of protein and nucleic acid synthesis have also been tested in the same system. Cytosine-arabinoside, an inhibitor of DNA synthesis, and actinomycin D, an inhibitor of m-RNA synthesis, have been shown to have no influence on the action of LH on testosterone synthesis. On the contrary, cycloheximide, a known inhibitor of protein synthesis, completely blocks the steroidgenic activity of LH as well as cyclic AMP. From these results, it is postulated that LH exerts its action on the testosterone biosynthesis through the formation of cyclic AMP, followed by rapid synthesis of unknown regulator protein.

抄録全体を表示

Triamcinolone acetonide (T-A) の筋注は臨床効果が長時間持続するが, その血中濃度の正確な報告はない.著者は本剤40mgをtriamcinolone acctonide4, 2, 4-³Hと共に筋注し血漿中のT-Aとtriamcinolone (T) 濃隻を経時的に測定した.その結果T-Aの有効濃度は筋注後14-21日間続くこと及びT-AよりTへの体内での転換は遅く, かつ少ないことを明らかにした.T-Aの尿中排泄率は7日間で16.5%であった.

抄録全体を表示

治療困難なインスリン抵抗性糖尿病患者にその抵抗性を除外する試みの一つとして蛋白分解酵素 (主としてα-chymotrypsin) を投与し, 血糖, 尿糖および全身状態の改善された症例を経験し, 有効例の臨床的観察を行なつた.さらに本酵素の血清に対する直接的影響をin vivo, in vitroにおける実験で検索し, 本酵素がインスリン阻害因子を抑制, 除去する一方インスリン作用と類似または協調する作用のあることを示唆する成績をえた.

抄録全体を表示

InsulinのRadioimmunoassayについて基礎的な検討を加え, これらを通してresin吸着法における最適条件を設定し, 更に手技の均一化と簡素化をる目的で作成されたFB-Separatorを導入して, 臨床検査法としての応用性について検討した.その成績より本法が膵内分泌のInsulin分泌機能の判定に応用しうることを認め, 更に本法を応用して糖尿病の各病期における膵内分泌機能の特性を示唆する所見を得た.

抄録全体を表示

Estrogenの下垂体FSH, LHに対するpositive feedback mechanismとClomid, F6066の作用機序の一端を解明する為, 雌性白鼠にest.benz,,premarinの大量1回投与時の下垂体FSH, LHの経時的変動及びClomid, F6066投与時の下垂体FSH, LHの経日的変動を分離定量した.est.benz., premarinl回大量投与すると短時間内に下垂体LH含量が増加し, Clomid, F6066投与では共に雌性白鼠下垂体FSH, LH含量が増加する事を明らかにした.

抄録全体を表示