Shinji Narukawa

Bcl-2 and Fas expression in eutopic and ectopic human endometrium during the menstrual cycle in relation to endometrial cell apoptosis

OBJECTIVE Our purpose was to investigate Bcl-2 and Fas expression in human eutopic and ectopic endometrium during the menstrual cycle in relation to endometrial cell apoptosis. STUDY DESIGN Eutopic and ectopic endometrial samples were obtained from 29 patients with endometriosis, and eutopic endometrial tissue samples were obtained from 9 patients with uterine myoma. Bcl-2 and Fas expression were examined by immunohistochemical staining with specific monoclonal antibodies; Bcl-2 expression in eutopic endometrium was also examined by Western blotting and apoptotic cells were detected by the labeling of deoxyribonucleic acid fragments. RESULTS In eutopic endometrium Bcl-2 was strongly expressed during the proliferative phase. Endometrial glandular cells showed evidence of cyclic changes in Bcl-2 expression, but cyclic changes were not apparent in peritoneal and ovarian endometriotic tissue. Fas expression was observed on glandular cells but not on stromal cells, and no cyclic changes in expression occurred in either ectopic or eutopic endometrium. Apoptotic cells were observed primarily in the glandular cells of the basal layer in eutopic endometrium during the late secretory and menstrual phases. CONCLUSION The current study indicated that there was no apparent correlation between Bcl-2 or Fas expression with endometrial cell apoptosis. The absence of cyclic changes in Bcl-2 expression in ectopic endometrium implied a difference in the mechanisms of proliferation or differentiation between eutopic and ectopic endometrium.

Monoclonal antiphosphatidylserine antibody reactivity against human first-trimester placental trophoblasts

OBJECTIVE To investigate the binding of antibodies against negatively charged phospholipids (antiphospholipid antibodies) to human placenta, we tested the reactivity of three mouse monoclonal antiphospholipid antibodies against first-trimester human placenta. STUDY DESIGN Formalin-fixed and frozen sections of first-trimester placentas were stained by immunoperoxidase with three mouse monoclonal antibodies. Each monoclonal antibody reacted differently with cardiolipin and phosphatidylserine, 3SB9b reacted with phosphatidylserine, D11A4 reacted with cardiolipin, and BA3B5C4 reacted with both. RESULTS 3SB9b reacted strongly with the syncytiotrophoblastic layer of both formalin-fixed and frozen placental tissue. Sporadic reactivity was observed against the cytotrophoblastic layer. BA3B5C4 reacted strongly and specifically with cytotrophoblastic cells. D11A4 reacted minimally or, more commonly, not at all. CONCLUSION The trophoblastic layer directly in contact with the maternal circulation is most reactive with antiphospholipid antibodies that react with phosphatidylserine rather than cardiolipin, suggesting that the trophoblasts may potentially be directly damaged by antiphospholipid antibodies through mechanisms unrelated to thrombosis. In addition, the differential reactivity of 3SB9b and BA3B5C4 suggests that the antigenic conformation involving phosphatidylserine on the cytotrophoblast is altered concurrent with fusion into the syncytium.

Hormonal Regulation in the Production of Macrophage Colony-Stimulating Factor and Transforming Growth Factor-Beta by Human Endometrial Stromal Cells in Culture

The effects of gonadal steroids on the secretion and gene expression of macrophage colony-stimulating factor (M-CSF) and on the secretion of transforming growth factor (TGF)-beta 1 and TGF-beta 2 by human endometrial stromal cells (ESCs) were examined by an in vitro system of ESC differentiation (decidualization). M-CSF production by ESCs was dose-dependently enhanced by the addition of progesterone or testosterone, while estradiol treatment had no effect. TGF-beta 2 secretion by ESCs was inhibited by progesterone, estradiol and testosterone treatment, and on the contrary, slight enhancement by estradiol was observed in TGF-beta 1 secretion. These findings indicate that human ESCs produce cytokines of M-CSF and TGF-beta s, which are important for the growth and differentiation of the peri-implantation embryo as well as local immune cells under direct control of gonadal steroidal actions, and suggest a novel network between endocrine and immune systems in the human endometrium.

Expression of messenger ribonucleic acid for gonadal steroid receptors in the human pelvic peritoneum

OBJECTIVE To investigate the expression of messenger RNA (mRNA) for gonadal steroid hormone receptors in the human pelvic peritoneum. DESIGN Analysis of estrogen receptor (ER), progesterone receptor (PR), and androgen receptor (AR) mRNA expressions in the pelvic peritoneum was carried out using the quantitative reverse transcription-polymerase chain reaction (PCR) method. SETTING Department of Gynecology and Obstetrics, Kyoto University Hospital, Kyoto, Japan. PATIENTS Pelvic peritoneal tissues from patients with (n = 10) and without (n = 10) endometriosis who had undergone gynecological surgery were studied. RESULTS Estrogen receptor, PR, and AR mRNAs were detected in all pelvic peritoneal samples analyzed. In the pelvic peritoneum of patients without endometriosis, ER mRNA levels were significantly lower in the luteal phase than in the follicular phase. This cyclic profile of ER mRNA expression was not observed in the pelvic peritoneum of patients with endometriosis. During the follicular phase, ER mRNA levels in the pelvic peritoneum of patients with endometriosis were significantly lower than those of patients with endometriosis. Neither PR nor AR mRNA levels in the pelvic peritoneum of either patient group showed significant cyclic variations throughout the menstrual cycle. A comparison of PR and AR mRNA levels in the pelvic peritoneum of the endometriosis and the nonendometriosis groups revealed no significant differences. CONCLUSIONS These data indicate a decrease in ER gene expression in the pelvic peritoneum of patients with endometriosis during the follicular phase. This suggests that the possible responsiveness of peritoneal cells to estrogen may be related to the occurrence and/or development of endometriosis.