Keisuke Wada

Peritoneal Fluid lnterleukin‐1β and Tumor Necrosis Factor in Patients With Benign Gynecologic Disease

ABSTRACT: The levels of interleukin 1β (IL‐1β) and tumor necrosis factor (TNF) in peritoneal fluid (PF‐IL‐1β and PF‐TNF) and production of IL‐1β and TNF by peritoneal macrophages were determined in patients with benign gynecologic disease. The level of PF‐IL‐1β was elevated in the acute pelvic inflammatory disease (PID) and stages I and II endometriosis (E I/II) groups compared with the normal pelvis group, but not in the myoma of the uterus, ovarian cyst, and postinflammatory pelvic adhesion groups. The level of PF‐TNF was elevated in the PID, EI/II and stages III and IV endometriosis (EIII/IV) groups. There was no correlation between the levels of PF‐IL‐β and PF‐TNF. Neither the level of PF‐IL‐β nor that of PF‐TNF was correlated with the concentration of peritoneal macrophages. Peritoneal macrophages produced IL‐1β and TNF in vitro in the absence of stimulants. The levels of PF‐IL‐1β and PF‐TNF are presumably linked to the activation of peritoneal macrophages.

Expression of interleukin-1 (IL-1) beta messenger ribonucleic acid (mRNA) and IL-1 receptor antagonist mRNA in peritoneal macrophages from patients with endometriosis

OBJECTIVE To investigate the expression of interleukin-1 (IL-1) beta messenger ribonucleic acid (mRNA) and IL-1 receptor antagonist (IL-1ra) mRNA in peritoneal macrophages. DESIGN, SETTING Peritoneal fluid (PF) samples were collected from patients who underwent laparoscopy or laparotomy. Northern blot analysis was performed at the reproductive research laboratory. PATIENTS Twenty-six patients with endometriosis, 10 patients with postinflammatory pelvic adhesion, and 12 control women with normal pelvis. MAIN OUTCOME MEASURE Polyadenylated RNA isolated from peritoneal macrophages was analyzed on Northern blots by using synthetic oligonucleotide probes. RESULTS The level of IL-1 beta mRNA expression was elevated in the group with stage I endometriosis, whereas the increased expression of IL-1ra mRNA was observed in the group with stages III and IV endometriosis. The level of IL-1 beta mRNA showed a positive correlation with that of IL-1 beta in PF and a negative correlation with the level of IL-1ra mRNA. CONCLUSIONS Our results suggest that peritoneal macrophages express IL-1ra mRNA rather than IL-1 beta mRNA with the progress of endometriosis and that peritoneal macrophages may secrete IL-1ra protein that modulates the effects of IL-1.

Danazol Suppresses the Production of Interleukin-1β and Tumor Necrosis Factor by Human Monocytes

ABSTRACT: The effects of estradiol (E2), progesterone (P), and danazol on the production of interleukin‐1β (IL‐1β) and tumor necrosis factor (TNF) by OK‐432 (a streptococcal preparation)‐stimulated monocytes were examined. E2 and P at physiologic concentrations enhanced IL‐1β and TNF production by monocytes from donors with lower control levels (without steroids added) of IL‐1β and TNF. However, E2 and P at physiologic concentrations did not affect IL‐1β and TNF production by monocytes from donors with higher control levels of IL‐1β and TNF. Danazol inhibited IL‐1β and TNF production by monocytes in a dose‐dependent manner from not only donors with lower control levels of IL‐1β and TNF but also donors with higher control levels of IL‐1β and TNF. Danazol at a concentration of 10−6 M significantly suppressed IL‐1β and TNF production in the presence of E2 and/or P at concentrations giving peak responses of IL‐1β production. These findings suggest possible new mechanisms of action for danazol in the treatment of endometriosis and infertility associated with immune abnormalities.

Estrogen binding sites in peripheral blood monocytes and effects of danazol on their sites in vitro

1. This study was designed to investigate the presence of estrogen type I (high affinity, low capacity) and type II (low affinity, high capacity) binding sites in human peripheral blood monocytes and the effects of danazol on these sites. 2. These two types of estrogen binding sites existed in human peripheral blood monocytes. 3. Danazol bound to these sites in high concentration (10(-6) M, clinical serum concentration during danazol therapy) and decreased the number of both sites. 4. It is suggested that danazol has an anti-estrogenic action to the monocytes through the competition and suppression of estrogen binding sites as seen in the estrogen target organ.

Different Effects on Oestrogen Binding Sites and Anti-Oestrogenic Action of Danazol and Progesterone

Rabbit uterus contains type I and II oestrogen binding sites in the cytosol, and nuclear fractions. Oestrogen-stimulated increase in uterine weight was inhibited by concurrent treatment with progesterone or danazol. Oestrogen-induced type I binding sites were decreased by progesterone and to a lesser extent by danazol, while oestrogen-induced type II sites were decreased almost equally in the two groups. Neither progesterone nor danazol alone caused any detectable changes in uterine weight. These findings may suggest that the anti-oestrogenic effect on uterine weight is more correlated with the change in type II binding sites of oestrogen than that in type I sites. Thus, danazol may exert anti-oestrogenic actions through a pathway independent from that of progesterone.

Rationale for frequency and dose of administration in gestrinone therapy for pelvic endometriosis in the experimental model of rabbit uterus.

1. Gestrinone has been used for treatment of pelvic endometriosis, in doses of 2.5 mg twice a week. This study is designed to clarify it from the dynamics of sex steroid receptors in rabbit uterus. 2. Four different regimens were scheduled, namely daily 1 microgram estradiol-17 beta (E2, consistent with endogenous level of women) for 3 days, and together with either 30, 60 (consistent with 2.5 microgram of clinical dose) or 120 microgram(s) gestrinone, in single dose or with 20 microgram(s) gestrinone daily (divided dose in single 60 microgram(s) gestrinone administration) for 3 days. Receptors for estrogen (ER, type I and II) or progestin (PR) were determined by charcoal adsorption in cytosol and KCl extract, or sedimentation in non-KCl extractable fraction, using [3H]E2 or [3H]promegestone respectively. 3. Gestrinone decreased total ER (type I and II) levels, dose-dependently, except the case of 20 micrograms gestrinone daily in ER type II. Total ER type I level did not return to the pre-level until 3 days only after 120 microgram(s) gestrinone administration. Total ER type II level was decreasing in 3 days after 30, 60 or 120 micrograms gestrinone therapy. Total PR level was decreased in the order of strength: 120 micrograms gestrinone greater than 60 micrograms gestrinone greater than 30 microgram gestrinone greater than 20 micrograms gestrinone in group, and the recovery was not obtained until 72 hr only after 120 micrograms gestrinone therapy. The uterine weight was decreasing with the same strength in 3 days of the therapy independently upon the regimen. 4. In conclusion, gestrinone dose of 60 micrograms (2.5 mg of clinical dose) every 3 days is considered to be effective for anti-steroid action in sex steroid receptor dynamics in the rabbit uterus, contributing to the rationale for gestrinone treatment of pelvic endometriosis.

Endocrine Features in Eutestosteronemic Women with Polycystic Ovaries

We attempted to assess the association between hyperandrogenemia and inappropriate gonadotropin secretion in women with polycystic ovaries (PCO). Thirty-one patients diagnosed as PCO by ultrasonography were divided into two subgroups: 17 with high serum total testosterone (T) level (> or = 0.5 ng/ml) and 14 with normal serum total T level (< 0.5 ng/ml). Both subgroups presented for the complaints of oligomenorrhea and/or hirsutism. The control group consisted of 15 women with regular ovulation for reference data collection. The PCO subjects with normal T, but not those with high T, revealed remarkable depletion of sex hormone binding globulin (SHBG), as compared with control. The PCO subject groups with high and normal serum T did not differ with respect to estrogen level, androgen level, follicle-stimulating hormone and prolactin levels, and SHBG concentration. Solely serum luteinizing hormone (LH) level was observed to be higher in those with high T, as typical features, than another subgroup or control. These data suggest that an increase in bioactive T as a result of decrease in serum SHBG or LH elevation may contribute to ovarian dysfunction in the patient with PCO.

Possible evidence for estrone-specific binding sites in human uterine endometrial carcinoma

Estrone (E1), a principal estrogen in postmenopausal women, may have profound consequences in the maintenance and progression of hormone-sensitive endometrial carcinoma. This study was designed to investigate the specific binding sites for E1 in the tumor and in the normal endometrium. The binding of [3H]E1 to the cytosolic fraction and KCl-soluble nuclear fraction from 3 endometrial carcinomas showed saturation kinetics with an apparent equilibrium dissociation constant of approximately 37 and 50 nM, respectively. The specific binding site of E1 was also found in 3 normal endometria. However, the binding capacity in the endometrial carcinoma increased to 1.5-fold of that in the normal endometrium. E2 did not affect [3H]E1 binding to any subcellular fraction from endometrial carcinoma specimens. These findings demonstrate the presence of specific binding sites for E1 in human endometrial carcinoma. The endometrial carcinoma growth may be mediated, at least in part, by E1 and its binding site interaction.