Ryohei Okamoto

Pharmacotherapeutic Effects of Kuei-chih-fu-ling-wan (Keishi-bukuryo-gan) on Human Uterine Myomas

Kuei-chih-fu-ling-wan (Keishi-bukuryo-gan; KBG), a traditional Chinese herbal remedy contains five components: bark of Cinnamomum cassia Bl. (Lauraceae), root of Paeonia lactiflora Pall. (Paeoniaceae), seed of Prunus persica Batsch. or P. persiba Batsch.var.davidiana Maxim. (Rosaceae), carpophores of Poria cocos Wolf. (Polyporaceae), and root bark of Paeonia suffruticosa Andr. (Paeoniaceae). This prescription has been frequently used in the treatment of gynecological disorders such as hypermenorrhea, dysmenorrhea and sterility. We previously reported that KBG might act as a LH-RH antagonist and a weak anti-estrogen on the uterine DNA synthesis in immature rats. In the present study, we investigated the effects of KBG on 110 premenopausal patients with uterine myomas. Clinical symptoms of hypermenorrhea and dysmenorrhea were improved in more than 90% of the cases with shrinking of uterine myomas in roughly 60% of the cases.

Differential gene expression of fibroblast growth factor receptor isoforms in rat ovary.

The gene expression of four fibroblast growth factor receptors (flg, bek, FGFR-3, and FGFR-4) in rat ovary cells was studied. Northern blot hybridization revealed that flg and bek mRNAs were detectable during all stages except a diestrus stage, whereas FGFR-3 and FGFR-4 mRNAs were almost undetectable throughout the cycles. In situ hybridization also demonstrated that only flg and bek gene expression was detectable. A modest flg mRNA signal was detected in developing antral follicles and it was more prominent in the theca-interstitial cells than in the granulosa cells. A modest to weak flg mRNA signal was seen in the hypertrophied theca-interstitial cells of atretic follicles and a very weak flg mRNA signal was observed in the corpora lutea. On the other hand, a weak bek mRNA signal was seen in granulosa and theca-interstitial cells in developing follicles and also hypertrophied theca-interstitial cells of atretic follicles, but not in the corpora lutea. Intense signals for both flg and bek mRNAs were unexpectedly found in the epithelium of paroophoron at the hilus. These results demonstrate that the bFGF receptor isoforms are expressed differentially in the rat ovary cells.

Effects of estrogen and progesterone on thymidine kinase activity in the immature rat uterus

The effects of progesterone and/or 17 beta-estradiol on thymidine kinase activity and autoradiograms were investigated in immature rats. Thymidine kinase activity increased more than thirtyfold above the control level 30 hours after 17 beta-estradiol injection. The enzyme activity induced by 17 beta-estradiol was suppressed by progesterone, the dose of which was approximately 1,000-fold that of 17 beta-estradiol. The specific thymidine kinase isozyme, which was separated from 17 beta-estradiol-induced uterine thymidine kinase by diethylaminoethyl (DEAE) cellulose column chromatography and not affected by deoxycytidine triphosphate, was involved in the DNA replication and inhibited by progesterone. The autoradiogram revealed many grains due to 3H-thymidine in the endometrial epithelium, stroma, and the myometrium in the immature rat 30 hours after 17 beta-estradiol injection, whereas progesterone reduced remarkably the number of grains induced by 17 beta-estradiol in the epithelium. Progesterone seems to inhibit the increment of the specific thymidine kinase isozyme induced by 17 beta-estradiol in the endometrial epithelium.

Anticancer effects of a chinese Herbal medicine, Juzen-taiho-to, in combination with or without 5-fluorouracil derivative on DNA-synthesizing enzymes in 1,2-dimethylhydrazine induced colonic cancer in rats

Juzen-taiho-to (JTT; [Shi-quan-da-bu-tang], a Japanese modified Chinese herbal prescription) in combination with an anticancer drug UFT (5-fluorouracil derivative) prevented the body weight loss and the induction of the colonic cancer in rats treated with a chemical carcinogen 1,2-dimethylhydrazine (DMH), and suppressed markedly the activity of thymidylate synthetase (TS) involved in the de novo pathway of pyrimidine synthesis in colonic cancer induced by DMH.

Differences in the induction of thymidine kinase isozymes in estrogen-treated immature and adult rats

Thymidine kinase activity in immature and castrated adult rat uterus has been examined in response to estrogen treatment. Following estrogen administration, it was found that immature uterine thymidine kinase activity was increased 30-fold after 24 h, but almost no effect was produced on castrated or non-castrated adult uterus. Uterine thymidine kinase activity was separated into three peaks (peak 1, 2 and 3) by means of DEAE-cellulose column chromatography. In response to estrogen, the thymidine kinase isozymes differed in adult and immature uteri. In immature uteri, marked and selective increase of the activity was found in peak I, whereas in adult only a slight increase in peak 2 activity was observed. The thymidine kinase activity in peak 1 and peak 2 were found to have different enzymatic properties and molecular weight, as determined by gel filtration of 125 000 for peak 1 and 100 000 for peak 2. From these results, it is suggested that estrogen induces specific thymidine kinase isozyme in immature uterus and that the isozyme may be involved in DNA synthesis. Such a induction mechanism seems to be lost during the development.

Basic fibroblast growth factor in rat corpus luteum stimulates prostaglandin F2-alpha production

Rat luteal cells (LC) were incubated with or without basic fibroblast growth factor (bFGF) in a serum-free medium. Prostaglandin F2 alpha (PGF2 alpha) production was stimulated in a dose-dependent manner at 0.03-1 ng/ml of bFGF. One ng/ml of bFGF caused approximately 5-fold the increment of PGF2 alpha at every stage of LC after 48 hrs of incubation. bFGF also raised progesterone secretion from LC, and this stimulatory effect on progesterone was more distinguishable in an early-, than a middle- and a late-stage. Additionally, bFGF concentration throughout the luteal phase was assessed using western blot analysis. The protein with typical molecular weight 18 kDa form was in high concentration throughout the luteal phase. These results suggest that bFGF may play a role in the regulation of PGF2 alpha and progesterone production as autocrine, but not in mitosis in corpus luteum.

Effects of estrogen and prolactin on thymidine kinase isozyme activities in DMBA-induced rat mammary tumor.

Effects of prolactin or estrogen on the activity of the pyrimidine salvage pathway enzyme thymidine kinase (TK) in 7,12-dimethylbenz(a)anthracene (DMBA)-induced mammary tumors were investigated in ovariectomized rats. Prolactin induced by perphenazine administration significantly enhanced the activities of total TK or its isozyme in the tumors. This specific TK isozyme separated by DEAE-cellulose column chromatography was suggested to be involved in DNA synthesis because it was not affected by dCTP. Its Mr was estimated to be approx. 100 000 by high-performance liquid chromatography. In contrast, estrogen alone did not raise the activity of the specific TK isozyme in the tumors.

Uremic Galactorrhea: an Endocrine Profile

Abnormal lactation (galactorrhea) was observed in 5 female patients with renal failure. This appeared in association with amenorrhea in the terminal stage of renal failure. Lactation gradually decreas

Effect of Ba-wei-di-huang-wan (Hachimi-jio-gan) on thymidine kinase and its isozyme activities in the prostate gland in rats

Ba-wei-di-huang-wan (Hachimi-jio-gan, TJ-7) has been frequently used for the treatment of sterilitas virilis, prostatic disorders, etc. The hormonal effects of TJ-7 were studied using thymidine kinase (TK), an enzyme that catalyzes the phosphorylation of thymidine via the pyrimidine salvage pathway. TK isozyme activity increases in rapidly proliferating tissues. TJ-7 may act as a weak androgen and enhance prostatic TK and in particular, a specific TK isozyme activity, and also as an anti-androgen and reduce prostatic TK activity in the presence of androgen in immature rats.

Effects of ACTH or hCG on corticosterone and progesterone secretion from adrenal glands of the rats at various ages

Effects of ACTH or hCG on the secretion of corticosterone (CB) and progesterone (PRG) from adrenal glands of the female rats at various ages were studied. In in vitro experiment, adrenal gland cells were isolated from fetal, neonatal, immature and adult rats by the method described by us before and were incubated in MEM. We chose to designate the hormones in the medium as hormone secretion. In in vivo experiment, hormones in the serum were determined. In vitro experiment, in which ACTH was added to the medium, indicated that CB was low in the media containing the cells from fetal and neonatal glands compared to those from immature and adult. PRG was elevated by ACTH addition at all of the ages. hCG addition had no effects on both CB and PRG in the media of immature and adult rats. In vivo experiment, in which ACTH was injected s.c., indicated that the response of serum PRG was slightly earlier than the CB response and peaked with a subsequent decline in immature rats. The results suggest that adrenal gland responds to ACTH from fetal age and secrets PRG, but little of CB in fetal age.