Soybean isoflavones inhibit estrogen-stimulated gene expression in mouse uteri.

This study was performed to examine the inhibitory effects of soybean isoflavones on estrogen-stimulated gene expression of the uteri in ovarectomized mice. Especially when compared with the inhibitory effect of genistein and daidzein as aglycosides described in our previous report, subcutaneous administration of the glycoside genistin significantly decreased the levels of estradiol-17beta (E2)-induced expressions of c-jun, interleukin (IL)-1alpha and tumor necrosis factor (TNF)-alpha mRNAs (p < 0.005, p < 0.05 and p < 0.05, respectively) and seemingly proteins in the mice uteri, whereas the glycoside daidzin weakly inhibited E2-stimulated expressions of c-fos and IL-1alpha. Both genistin and daidzin seemed to have a weaker inhibitory effect than that of genistein and daidzein on the expression of estrogen-stimulated genes. It is suggested that those glycosides are naturally derived and generally absorbed from plant foods and might prevent E2-related endometrial carcinogenesis.

Preventive effects of Juzen-taiho-to on N-methyl-N-nitrosourea and estradiol-17beta-induced endometrial carcinogenesis in mice.

Two experiments were conducted to determine effects of Juzen-taiho-to on endometrial carcinogenesis in mice. In the first experiment, Juzen-taiho-to treatment (2 weeks) decreased the levels of estradiol-17beta (E(2))-stimulated expression of c-fos/jun mRNA and their oncoproteins, determined by reverse transcription-polymerase chain reaction and Southern blot analysis, and the immunohistochemical method, in the uteri of ovarectomized mice. For the second experiment, 93 female ICR mice were given N-methyl-N-nitrosourea (MNU) solution (1 mg/100 g body weight) and normal saline (as controls) into their left and right uterine corpora, respectively, and were divided into four groups. Group 1 was given a diet containing 0.2% Juzen-taiho-to and 5 p.p.m. E(2). Group 2 was given a diet containing 5 p.p.m. E(2) alone. Group 3 was given a diet containing 0.2% Juzen-taiho-to alone. Group 4 was kept on the basal diet alone and treated as a control. Juzen-taiho-to treatment significantly decreased incidences of the uterine endometrial atypical (P<0.01), complex (P<0.05) and simple hyperplasias (P<0.01), under estrogenic stimulation. It is suggested that Juzen-taiho-to has an inhibitory effect on E2-related endometrial carcinogenesis in mice, relevantly through suppression of estrogen-induced c-fos/jun-expression.

Herbal complex suppresses telomerase activity in chemo-endocrine resistant cancer cell lines.

A herbal complex consisting of Hoelen, Angelicae radix, Scutellariae radix and Glycyrrhizae radix suppressed cell viability and telomerase activity in hormone-refractory and chemo-resistant cancer cell lines, namely poorly differentiated uterine endometrial cancer cell line AN3 CA, adriamycin-resistant breast cancer cell line MCF7/ADR and cisplatin-resistant ovarian cancer cell line A2780. Furthermore, the herbal complex suppressed the expression of the full length of human telomerase reverse transcriptase (hTERT), which is related to telomerase activity. This indicates that the herbal complex can suppress the tumor growth of chemoendocrine resistant cancers, at least in part via suppression of telomerase activity associated with down-regulated hTERT.

Preventive effects of Glycyrrhizae radix extract on estrogen-related endometrial carcinogenesis in mice.

Short- and long-term experiments were conducted to examine the effects of Glycyrrhizae radix (Gl radix) extract on mouse endometrial carcinogenesis. Gl radix treatment (2 weeks) decreased the levels of c-fos/jun mRNA and the corresponding oncoproteins induced by estradiol-17 beta (E2) in castrated mice uteri, as determined by reverse transcription-polymerase chain reaction and Southern blot analysis, and immunohistochemical methods, respectively. For the long-term assays, 98 female ICR mice were given N-methyl-N-nitrosourea (MNU) solution (1 mg/100 g body wt.) and normal saline (as controls) into their left and right uterine corpora, respectively. They were divided into four groups as follows: group 1 was given 0.625% Gl radix- and 5 ppm E2-containing diet; group 2, 5 ppm E2-containing diet; group 3, 0.625% Gl radix-containing diet; and group 4, the basal diet alone. Gl radix treatment significantly decreased uterine weights and the incidences of uterine endometrial atypical hyperplastic and malignant lesions. It is suggested that Gl radix has inhibitory effects on E2-related endometrial carcinogenesis in mice, through suppression of estrogen-induced c-fos/jun-expressions.

Therapeutic effects of an injectable new quinolone, pazufloxacin, against polymicrobial infections in the uterine endometritis model.

Polymicrobial infections with aerobes and anaerobes are common in female genital tract infections. We evaluated the efficacy of an injectable new quinolon, pazufloxacin, using a uterine endometritis model. Rats were infected with a mixed inoculation of Escherichia coli plus Bacteriodes fragilis (MIC of pazufloxacin and ceftazidime: E. coli: 0.05 and 1.56 micrograms/ml, respectively, B. fragilis: 3.13 and 3.13 micrograms/ml, respectively). After inoculating 10(7) cfu/rat of each organism, pazufloxacin or ceftazidime (10 or 20 mg/kg, respectively, i.v., b.i.d., 3 days) was administered and compared with the nontreated group. The viable cell counts of the uterine corpus and uterine cervix in pazufloxacin-treated and ceftazidime-treated groups were decreased, compared with the nontreated group. The viable cell counts of the adnexa in the pazufloxacin-treated group were significantly decreased, compared with the ceftazidimetreated group. These results suggest that pazufloxacin would be useful for the treatment of polymicrobial infections, especially adnexitis.

Plausible novel therapeutic strategy of uterine endometrial cancer with reduction of basic fibroblast growth factor secretion by progestin and O-(chloroacetyl-carbamoyl) fumagillol (TNP-470; AGM-1470).

To know the potential of growth, invasion and metastasis of endometrial cancer associated with neovascularization, the effects of sex steroids and O-(chloroacetyl-carbamoyl) fumagillol (TNP-470; AGM-1470) on basic fibroblast growth factor (FGF) expression and secretion and its mRNA expression were investigated in well-differentiated endometrial cancer cell line Ishikawa and in undifferentiated endometrial cancer cell line AN3 CA. Basic FGF expression and secretion and its mRNA expression in Ishikawa cells, but not in AN3 CA cells, were increased by estrogen, while progesterone diminished the estrogen-induced increases. TNP-470 reduced the levels regardless of estrogen treatment in AN3 CA cells. Therefore, basic FGF secretion may be inhibited by progestin in differentiated cells, and by TNP-470 in undifferentiated cells. Since endometrial cancer consists of differentiated and undifferentiated cells as heterogeneity, a combination therapy for endometrial cancer with progestin and TNP470 might be effective.

Comparative study on vaginal or oral treatment of bacterial vaginosis.

The bacterial epidemiology of bacterial vaginosis (BV) and the efficacy of vaginal or oral treatment of BV with clindamycin (CLDM) were investigated. The epidemiology of BV was investigated in 100 symptomatic women before CLDM therapy. Two groups consisting of 50 patients each with the diagnosis of symptomatic BV were treated with either oral administration of 450 mg CLDM three times daily or 2% CLDM phosphate in vaginal cream (self-made) 5 g once a day, for 7 days. There was no significant differences in efficacy among vaginal and oral therapies with CLDM. Vulvovaginal irritation occurred in 3 patients orally treated and in 1 patient vaginally treated. Gastrointestinal disturbances were observed in 4 orally treated patients. A slight abnormal elevation of the glutamine-oxaloacetic transaminase level was also found in 1 patient orally treated. Since there were no statistically significant differences in efficacy rates between vaginal and oral CLDM treatments, we favor vaginal treatment of BV, based on less adverse effects.

[Effect of preoperative oral 5-fluorouracil on cell cycle of advanced cervical squamous cell carcinoma].

Using flow cytometry, we examined the correlations between the tumor stage and the cell cycle in subjects with advanced cervical squamous cell carcinoma as well as between the changes of cell cycle caused by 5-FU and their prognoses. Before 5-FU treatment, 75.9 +/- 9.6% of the patients were at G1 phase, 16.8 +/- 3.9% at S and 7.3 +/- 4.5% at G2/M. The change of cell cycle was not accompanied by the progression of tumor stage. Sixteen patients received oral 5-FU (300 mg/day) for at least 2 weeks before operation. In the patients at stage I, the cell cycle was not changed even after 5-FU treatment. They have been cancer-free for 3 postoperative years. While early recurrence was found in 4 of 5 cases at stage II or III showing progression at G1, all 6 cases who had regressed or remained unchanged at G1 were free from recurrence (p = 0.0152). These results suggested that the cell cycle is not necessarily correlated with the progression of tumor stage in patients with this kind of carcinoma, and that the change of cell cycle due to preoperative oral 5-FU could be a predictive indicator for their prognoses. Especially for those with poor prognoses who had shown progression at G1, more potent adjuvant chemotherapy should be planned.

Possible evidence that the herbal medicine shakuyaku-kanzo-to decreases prostaglandin levels through suppressing arachidonate turnover in endometrium.

The herbal medicine shakuyaku-kanzo-to has been used for the treatment of dysmenorrhea. We attempted to establish the effects of shakuyaku-kanzo-to on prostaglandin (PG) levels and the turnover of arachidonic acid in endometrial cells. Human endometrial cells isolated from proliferative-phase endometria were examined. Arachidonic acid reacylation activity was determined by [3H]arachidonic acid incorporation into endometrial cell phospholipid. Its deacylation rate was determined by both 3H-liberation from [3H]-arachidonic acid-labelled phospholipids and production of lyso-phospholipid. [3H]-PG levels were also analyzed. Shakuyaku-kanzo-to enhanced the rate and maximal level of arachidonic acid incorporation by 1.5-fold into phosphatidyl-ethanolamine and phosphatidylcholine. Degradation of the phospholipids to liberate arachidonic acid and lyso-derivatives was not affected by the shakuyaku-kanzo-to. Shakuyaku-kanzo-to decreased PG levels by approximately 50% of control. Because shakuyaku-kanzo-to stimulates arachidonic acid esterification to phospholipids, it seems likely that shakuyaku-kanzo-to depletes cellular free arachidonic acid with a consequent suppression of prostaglandin synthesis. Shakuyaku-kanzo-to may exert its action against dysmenorrhea through preventing prostaglandin production.

Study on treatment of bacterial vaginosis with oral administration of metronidazole or cefdinir.

Bacterial vaginosis (BV) is considered to be one of the most common vaginal infections in women. Fifteen symptomatic women with BV were enrolled in this study. Ten patients with the diagnosis of BV were treated with 10 days oral administration of metronidazole (MTN), 500 mg twice a day, and five patients with cefdinir (CFDN), 300 mg three times a day. In the MTN therapy, the rate of abnormal vaginal discharge subjectively decreased from 100 to 60%, the rate of abnormal vaginal discharges objectively decreased from 100 to 20%, the rate of positive amine tests decreased from 100 to 20%, the rate of genital malodor and abnormal pH of vaginal discharges decreased substantially from 100 to 10%, and the rate of the presence of clue cells also decreased notably from 90 to 10%. However, in the CFDN therapy, none of these factors improved. With respect to susceptibility to CFDN and MTN, CFDN demonstrated good antibacterial activity against almost all bacteria isolated except Gardnerella vaginalis. On the other hand, MTN demonstrated excellent activity against anaerobic bacteria except Peptostreptococcus spp., and had no antibacterial activity against aerobic bacteria. Since the therapeutic effect of MTN in BV appeared to be better than that of CFDN, anaerobes may play a major role in causing clinical symptoms in patients with BV.

Possible mechanism of steroid action of the plant herb extracts glycyrrhizin, glycyrrhetinic acid, and paeoniflorin: inhibition by plant herb extracts of steroid protein binding in the rabbit.

To assess the action of some components of herbal medicine, glycyrrhizin, glycyrrhetinic acid, and paeoniflorin, on steroids, their binding to several classes of intracellular and serum steroid-binding proteins were studied in the rabbit. The affinity (inhibitor constant) for binding of dihydrotestosterone-sex hormone binding globulin in the serum (dissociation constant of 2.0 nmol/L for dihydrotestosterone) was approximately 520 nmol/L, and that for the binding of cortisol-corticosteroid-binding globulin in the serum (dissociation constant of 2.0 nmol/L for cortisol) was approximately 10 mumol/L. In the uterine cytosol, the inhibitor constant value for estradiol receptor binding (dissociation constant of 1.0 nmol/L) was 0.9 mumol/L, and these compounds did not influence progestin receptor binding (dissociation constant of 1.4 nmol/L). The inhibitor constant values for glucocorticoid receptor binding (dissociation constant of 1.0 nmol/L) in the liver cytosol were 3.0 nmol/L for paeoniflorin, 2.0 nmol/L for glycyrrhizin, and 1.7 nmol/L for glycyrrhetinic acid, and those for mineralocorticoid receptor binding (dissociation constant = 1.1 nmol/L) in the kidney cytosol were 3.5 nmol/L for paeoniflorin and glycyrrhetinic acid and 3.0 nmol/L for glycyrrhizin. These results suggest that herbal extracts such as the above compounds influence steroid effects by glucocorticoid and mineralocorticoid receptors and to a lesser extent by estrogen receptors or serum sex hormone-binding globulin and corticosteroid-binding globulin.

Inhibition by plant herb extracts of steroid bindings in uterus, liver and serum of the rabbit.

To assess the action of herb extracts, glycyrrhizin and paeoniflorin, upon steroids, their binding to several classes of intracellular and serum steroid-binding proteins was studied in the rabbit. Glycyrrhizin and paeoniflorin exhibited similar binding behaviors. They bound minimally to estrogen and androgen receptors, but not to the progesterone receptor in uterine cytosol, and exhibited a moderate binding activity to glucocorticoid receptors in liver cytosol. They also exhibited weak binding activity to both cortico-steroid-binding globulin and sex-hormone-binding globulin.