Tomio Iwabe

Role of cytokines in endometriosis-associated infertility.

Endometriosis, which is common in women of reproductive age, may affect fertility. It is also clear that mechanical disruption of the pelvic anatomy may cause infertility. However, our understanding of the association between the early stage of endometriosis and infertility remains incomplete. Bloody peritoneal fluid (PF) is frequently observed in the cul-de-sac of endometriosis patients and contains various biologically active factors. We found that the concentrations of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) in PF from patients with endometriosis were significantly higher than that of patients with endometriosis. There were significantly positive correlations between the levels of TNF-α and IL-6. We compared the levels of these cytokines with regard to the R-AFS stages and scores, but no differences were observed. In contrast, these cytokines correlate with the number and extent of red color peritoneal endometriosis. TNF-α increased the expression of IL-6 messenger RNA and protein in endometriotic stromal cells derived from chocolate cyst in a dose-dependent manner. The addition of IL-6 inhibited the development of mouse preimplantation embryo and impaired sperm function. We concluded that increased levels of IL-6 in peritoneal fluid of patients with active red endometriosis might be related to endometriosis-associated fertility.

Apoptosis and endometriosis.

Apoptosis is a distinctive form of programmed cell death resulting in the efficient elimination of cells without eliciting an inflammatory response. Endometriosis is characterized by the presence of endometrial cells with capacity to avoid apoptosis outside the uterus. Apoptosis plays a fundamental role for the pathogenesis of endometriosis. Eutopic endometrium from women with endometriosis has increased expression of anti-apoptotic factor and decreased expression of pro-apoptotic factors compared with endometrium from healthy women. These differences could contribute to the survival of regurgitating endometrial cells into the peritoneal cavity and development of endometriosis. Increased apoptosis of Fas-bearing immune cells in the peritoneal cavity may leads to their decreased scavenger activity that eventually results in prolonged survival of ectopic endometrial cells in women with endometriosis. This study is a current review of the literatures focused on the physiological role of apoptosis in normal endometrium and alterations in regulation of apoptosis in eutopic and ectopic endometrium from women with endometriosis. The role of apoptosis in the treatment of endometriosis is also reviewed.

Subtle rise in serum progesterone during the follicular phase as a predictor of the outcome of in vitro fertilization

OBJECTIVE To investigate the effects of subtle rises in serum progesterone (P) during the follicular phase on the outcome of in vitro fertilization and embryo transfer (IVF-ET). DESIGN, PATIENTS One hundred one patients underwent IVF-ET for 170 cycles and were stimulated with a combination of clomiphene citrate and human menopausal gonadotropin. Based on their hormonal data, we divided the patients into two groups: those who had a cycle with an increase in serum P concentration (1.0 to 2.0 ng/mL) that was not associated with a pituitary LH release (subtle P rise) and those who had a cycle without any increase in serum P concentration (no P rise). MAIN OUTCOME MEASURES The daily serum estradiol (E2) concentration and the results of IVF-ET (number of developed and collected oocytes, rates of mature oocytes, fertilization, and pregnancy) were compared between the two groups. RESULTS Subtle P rises were observed in 31.7% (32/101) of the patients and 20.5% (36/170) of the cycles evaluated during the IVF-ET programs. A significantly higher serum E2 concentration (P less than 0.001) and a greater number of developed and collected oocytes (P less than 0.001 and P less than 0.05, respectively) also were observed in those cycles with a subtle P rise. The rates of mature oocyte formation and fertilization were significantly lower in cycles with a subtle P rise (P less than 0.001 and P less than 0.05, respectively). A lower pregnancy rate was observed in cycles with a subtle P rise, and all 12 ongoing pregnancies occurred only in cycles with a no P rise. CONCLUSION These results suggest that the development of an increased number of follicles may not necessarily improve the outcome of IVF-ET and that the measurement of serum P may be a better predictor for successful pregnancy.

Gonadotropin-releasing hormone agonist treatment reduced serum interleukin-6 concentrations in patients with ovarian endometriomas

OBJECTIVE To determine whether serum interleukin (IL)-6 can be measured in patients with ovarian endometriomas and whether these measurements are useful in managing this disease. DESIGN A controlled clinical study and an in vitro study. SETTING Department of Obstetrics and Gynecology, Tottori University, Japan.Twenty-two patients with ovarian endometriomas. INTERVENTION(S) Laparoscopic cystectomy for ovarian endometriomas was performed. Gonadotropin-releasing hormone (GnRH) agonist was administered for 3 months in nine patients before laparoscopic surgery. Endometriotic stromal cells obtained from patients with endometriomas with or without GnRH agonist treatment were cultured. MAIN OUTCOME MEASURES(S) IL-6 concentrations in serum or supernatant of the cell culture were measured using ELISA. RESULTS The serum concentration of IL-6 in patients with endometriomas was higher at the time of diagnosis than in those without endometriomas. Laparoscopic surgery significantly reduced serum levels of IL-6. Serum IL-6 concentrations also decreased after treatment with GnRH agonist. IL-6 production was attenuated in the endometriotic stromal cells obtained from patients with GnRH agonist treatment compared with patients without such treatment. CONCLUSION(S) GnRH agonist treatment may decrease IL-6 production in endometriotic cells. Measurement of serum IL-6 concentrations may be of value in managing patients with endometriomas.

Adenomyosis in a patient with the Rokitansky-Kuster-Hauser syndrome

Endometriosis remains an enigma despite extensive clinical investigations and experience. Several theories have attempted to explain the puzzling and controversial pathogenesis of endometriosis. Sampson’s implantation theory is the most commonly accepted. Early endometriotic lesions have been found in the Douglas cavity, where retrograde menstruation was confirmed in 90% of women. These findings support Sampson’s theory. In contrast, endometriosis occurs in some patients who have no functional endometrium, such as those with the Rokitansky-Kuster-Hauser syndrome, a clinical condition that supports Iwanoff and Mayer’s coelomic metaplasia theory.

Serum interleukin-8 levels are elevated in patients with ovarian endometrioma

OBJECTIVE To determine whether serum interleukin (IL)-8 concentration can be measured in patients with ovarian endometrioma and whether this measurement is a useful tool in diagnosing this disease. DESIGN A controlled clinical study and an in vitro study. SETTING Department of Obstetrics and Gynecology, Tottori University, Japan. PATIENT(S) Seventy patients with ovarian endometrioma and 21 patients with benign ovarian cyst. INTERVENTION(S) Laparoscopic surgery or laparotomy for ovarian endometriomas or benign ovarian cyst was performed. Preoperative blood samples were obtained. Endometriotic stromal cells obtained from nine patients with endometrioma were cultured. MAIN OUTCOME MEASURE(S) Interleukin-8 concentration in the serum or supernatant of the cell culture was measured with use of ELISA. RESULT(S) The serum concentration of IL-8 in patients with endometrioma was significantly higher than in patients with benign ovarian cyst. The serum IL-8 threshold (25 pg/mL) had a higher sensitivity (71.4%) for diagnosing ovarian endometrioma than did serum CA-125 level. The increased rates of IL-8 concentration in the culture supernatants after adding tumor necrosis factor alpha were significantly higher in patients whose serum IL-8 levels were >or=25 pg/mL than in those with levels <25 pg/mL. CONCLUSION(S) Measuring of serum IL-8 concentration may be a valuable tool in diagnosing endometriosis.

TAK1 activation for cytokine synthesis and proliferation of endometriotic cells

Endometriosis causes pelvic pain and infertility in women of reproductive age. We explored TNFalpha-induced specific signaling pathways and gene expressions in endometriotic stromal cells (ESCs). Based on the data of the pathway specific cDNA array, we analyzed the role of TAK1, which is believed to work as a common mediator for NF-kappaB and MAPK pathways. Using the NF-kappaB pathway array, we found that TNFalpha upregulated ICAM-3, IL-6, IL-8, TAK1, JNK2, RelA, and TLR4 expressions. TNFalpha augmented the phosphorylation of TAK1. By transfection of TAK1 siRNA, TNFalpha-induced phosphorylation of IkappaBalpha, JNK1/2, and p38MAPK, as well as IL-6 or IL-8 expression, were repressed. TAK1 silencing in TNFalpha-pretreated ESCs caused a decrease in the proportion of cells in S-phase, and reduced TNFalpha-promoted BrdU incorporation. We provide the first evidence that TNFalpha and its downstream TAK1, which are key mediators for NF-kappaB and MAPK pathways, may be involved in the pathogenesis of endometriosis.

Interleukin-1beta-induced expression of IL-6 and production of human chorionic gonadotropin in human trophoblast cells via nuclear factor-kappaB activation.

Problem:  The cytokine, interleukin‐6 (IL‐6), stimulates the production of human chorionic gonadotropine (hCG) in chorionic cells. The purpose of this study was to examine the role of nuclear factor‐κB (NF‐κB) during the induction of IL‐6 by IL‐1β in human trophoblast cells.

Interleukin-8 gene and protein expression are up-regulated by interleukin-1β in normal human ovarian cells and a granulosa tumor cell line

OBJECTIVE To evaluate the expression, regulation, and role of interleukin (IL)-8 in human ovary. DESIGN Prospective study. SETTING University hospital. PATIENT(S) Sixteen premenopausal women. INTERVENTION Follicular fluid and granulosa lutein cells (GLCs) were collected during IVF cycles. Ovarian stromal and theca cells were obtained from women underwent surgery. KGN cells, the human granulosa cell tumor cell line, were also used. MAIN OUTCOME MEASURES The levels of IL-8 and IL-1beta in follicular fluid and IL-8 protein production were determined using ELISA. Interleukin-8 and IL-8 receptor gene expression in ovarian cells and the effect of IL-8 on the proliferation of stromal cells were determined. The expression of pIkappaB was evaluated by Western blot, and the effect of NF-kappaB inhibitor APDC was examined by Northern blot analysis and ELISA in KGN cells. The levels of IL-8 and IL-1beta in follicular fluid; each concentration and the volume showed a positive correlation. Reverse transcription polymerase chain reaction showed the presence of IL-8 mRNA in all ovarian cells. In contrast, IL-8 receptor mRNA was only detected in stromal cells. The expression of IL-8 in GLCs and KGN cells was increased by addition of IL-1beta and TNFalpha. Interleukin-8 increased the proliferation of ovarian stromal cells. The expression of pIkappaB in KGN cells was induced by IL-1beta, and the effects were reduced by APDC. CONCLUSION(S) Interleukin 8 induced by IL-1beta via activation of NF-kappaB in granulosa cells may have a role in the periovulatory period of follicular maturation.

The Importance of Venous Doppler Velocimetry for Evaluation of Intrauterine Growth Restriction

The management of growth‐restricted fetuses requires accurate diagnosis to optimize the timing of delivery. Doppler velocimetry is the only noninvasive method for assessing the fetoplacental hemodynamic status. This review will give a critical overview of the current knowledge on fetal venous blood flow in pregnancies complicated by in‐trauterine growth‐restricted fetuses. Adaptation of the circulation in intrauterine growth‐restricted fetuses is described. Normal and abnormal venous Doppler waveforms are presented. Correlations of abnormal waveforms with the presence of acidemia and perinatal outcomes are emphasized. Limitations of venous Doppler velocimetry for optimizing the time for delivery and the perinatal outcome are also presented.