Atsuko Wakabayashi

Inhibitory effect of curcumin on uterine leiomyoma cell proliferation

Objective. Uterine leiomyomas are the most common gynaecological benign tumour and greatly affect reproductive health and wellbeing. They are the predominant indication for hysterectomy in premenopausal women. Curcumin, a well-known component of turmeric, has been reported to prevent various diseases such as cancer, diabetes and obesity. Previous study reported that curcumin represses the proliferation of several tumour cells. However, there has not been a precise characterisation of the curcumin-induced inhibition of uterine leiomyoma cells. In this study, we investigated the inhibitory effect of curcumin on leiomyoma cells proliferation. Study design. Eker rat-derived uterine leiomyoma cell lines (ELT-3 cells) were used. Cell proliferation was assessed by counting the number of cells and MTS assay. The activation of peroxisome proliferator-activated receptor-gamma (PPARγ) was evaluated by luciferase assay. Results. We found that curcumin significantly inhibited ELT-3 cell proliferation. PPARγ was expressed in ELT-3 cells and curcumin acted as a PPARγ ligand. This inhibitory effect of curcumin was attenuated by the treatment of cells with PPARγ antagonist. Conclusion. These experimental findings in vitro show that the inhibitory effect of curcumin on ELT-3 cell proliferation occurs through the activation of PPARγ. Curcumin may be useful as an alternative therapy for uterine leiomyoma.

Balloon tamponade during cesarean section is useful for severe post-partum hemorrhage due to placenta previa

Aim:  Severe post‐partum hemorrhage during cesarean section due to placenta previa is still one of the leading causes of maternal mortality. The aim of this study was to evaluate the efficiency of intrauterine tamponade with a Sengstaken‐Blakemore tube (SB‐tube) for the treatment of severe post‐partum hemorrhage in cases of placenta previa.

Targeting interleukin-6 receptor inhibits preterm delivery induced by inflammation

Intrauterine infection is still a common trigger of preterm delivery (PTD) and also a determinant risk factor for the subsequent development of neurodevelopmental abnormalities in neonates. In this study, we examined the expressional pattern of various inflammatory cytokines such as interleukin-1β (IL-1β), IL-6 and tumor necrosis factor-α (TNF-α) in placentae complicated with severe chorioamnionitis (CAM) and found that IL-6 is mainly expressed in macrophages in villous mesenchyme by immunohistochemical analysis with anti-CD-68 antibody. Using an experimental lipopolysaccharide (LPS)-induced PTD model, the therapeutic potential of targeting this cytokine was investigated. Anti-IL-6 receptor antibody (MR16-1) was delivered 6 h before LPS treatment. Mice in the MR16-1 group had a significantly lower rate of PTD (17%) than in the controls (53%, P = 0.026). As a result, MR16-1 treatment significantly prolonged the gestational period (control; 18.4 ± 1.7d, MR16-1; 19.8 ± 1.5d, P = 0.007) without any apparent adverse events on the mice and their pups. In primary human amniotic epithelial cells, pretreatment with a humanized anti-human IL-6 receptor antibody, tocilizumab, significantly inhibited the production of prostaglandin E2 induced by IL-6. In conclusion, IL-6 was strongly expressed mainly in macrophages in villous mesenchyme in placentae complicated with CAM. Anti-IL-6R antibody significantly decreased the rate of PTD in LPS-induced inflammatory model in mice, and inhibited PGE2 production from human primary amniotic epithelial cells. Targeting IL-6 signaling could be a promising option for the prevention of PTD and needs to be further explored for future clinical application.

Essential Function for the Nuclear Protein Akirin2 in B Cell Activation and Humoral Immune Responses

Akirin2, an evolutionarily conserved nuclear protein, is an important factor regulating inflammatory gene transcription in mammalian innate immune cells by bridging the NF-κB and SWI/SNF complexes. Although Akirin is critical for Drosophila immune responses, which totally rely on innate immunity, the mammalian NF-κB system is critical not only for the innate but also for the acquired immune system. Therefore, we investigated the role of mouse Akirin2 in acquired immune cells by ablating Akirin2 function in B lymphocytes. B cell–specific Akirin2-deficient (Cd19Cre/+Akirin2fl/fl) mice showed profound decrease in the splenic follicular (FO) and peritoneal B-1, but not splenic marginal zone (MZ), B cell numbers. However, both Akirin2-deficient FO and MZ B cells showed severe proliferation defect and are prone to undergo apoptosis in response to TLR ligands, CD40, and BCR stimulation. Furthermore, B cell cycling was defective in the absence of Akirin2 owing to impaired expression of genes encoding cyclin D and c-Myc. Additionally, Brg1 recruitment to the Myc and Ccnd2 promoter was severely impaired in Akirin2-deficient B cells. Cd19Cre/+Akirin2fl/fl mice showed impaired in vivo immune responses to T-dependent and -independent Ags. Collectively, these results demonstrate that Akirin2 is critical for the mitogen-induced B cell cycle progression and humoral immune responses by controlling the SWI/SNF complex, further emphasizing the significant function of Akirin2 not only in the innate, but also in adaptive immune cells.

Antiproliferative effect of adiponectin on rat uterine leiomyoma ELT-3 cells

Objective. Although fibroids greatly affect reproductive health, the pathophysiology and epidemiology are not well known. Recently, we have reported the relationship between uterine leiomyoma and metabolic syndrome. Many studies have indicated that reductions in plasma adiponectin levels play major roles in the development of metabolic syndrome. In this study, we investigated the significant repressive effect of adiponectin on rat uterine leiomyoma ELT-3 cells proliferation. Study design. Expression of adiponectin receptor 1 and receptor 2 was evaluated by RT-PCR and Western blot analysis. Cell proliferation was assessed by the MTS assay and cell counting. Apoptosis was evaluated by Hoechst staining and cleaved poly (ADP-ribose) polymerase (PARP). Results. Adiponectin receptor 1 and receptor 2 were expressed in ELT-3 cells. Adiponectin repressed rat uterine leiomyoma ELT-3 cells cell proliferation without inducing apoptosis. Conclusion. The repression of adiponectin on leiomyoma cell proliferation in the rat may explain a crucial role of adiponectin in the association of metabolic syndrome with uterine leiomyoma.

Aldosterone stimulates the proliferation of uterine leiomyoma cells.

Objective. Although uterine leiomyomas are the most common gynaecological benign tumour and greatly affect reproductive health and well being, the pathophysiology and epidemiology of uterine leiomyomas are not well known. Elevated blood pressure has an independent, positive association with risk for clinically detected uterine leiomyomas. Aldosterone is a key biological peptide in the renin-angiotensin-aldosterone system that regulates blood pressure. In this study, we investigated the siginificant stimulatory effect of aldosterone on leiomyoma cells proliferation. Study design. This study investigated the potential role of aldosterone in the proliferation of ELT-3 leiomyoma cells. Results. Aldosterone-induced ELT-3 leiomyoma cell proliferation and the expression of mineralocorticoid receptor (MR) were confirmed. Pre-incubating the cells with the MR blockers spironolactone or eplerenone effectively repressed aldosterone-induced and angiotensin II (Ang II)-induced cell proliferation. Treatment of aldosterone increased the levels of Ang II type-1 receptor mRNA. Conclusion. These experimental findings in vitro show the presence of complex regulation of Ang II and aldosterone induced leiomyoma cell proliferation.

Repressive effect of the phytoestrogen genistein on estradiol-induced uterine leiomyoma cell proliferation.

Objective. Uterine leiomyomas are the most common gynecological benign tumor and greatly affect reproductive health and well-being. They are the predominant indication for hysterectomy in premenopausal women. Current epidemiological study reported that soy products intake is inversely associated with diseases leading to hysterectomy. Genistein is a soy-derived phytoestrogen and its inhibitory effect on leiomyoma cell proliferation is reported. In this study, we investigated the siginificant inhibitory effect of genistein on estradiol (E2)-induced leiomyoma cells proliferation. Study design. The Eker rat-derived uterine leiomyoma cell line ELT-3 cells were used. Cell proliferation was assessed by counting the number of cells. The expression of estrogen receptors and peroxisome proliferator-activated receptor-γ (PPARγ) was evaluated by Western blot analysis. Results. PPARγ was expressed in ELT-3 cells and genistein acted as PPARγ ligand. This inhibitory effect of genistein was attenuated by the treatment of cells with PPARγ antagonist bisphenol A diglycidyl ether (BADGE) or GW9662. Conclusion. These experimental findings in vitro show that the repressive effect of genistein on E2-induced ELT-3 cell proliferation is through the activation of PPARγ. Genistein may be useful as an alternative therapy for leiomyoma.

Estimation of mouse fetal weight by ultrasonography: application from clinic to laboratory:

Ultrasonographic assessment of fetal growth to estimate fetal weight has been widely used in clinical obstetrics but not in laboratory mice. Even though it is important to assess fetal growth abnormalities for gene-targeting studies using mice, there have been no reports of accurately estimated fetal weight using fetal biometric parameters in mice. The aim of this study was to establish an accurate mouse formula using fetal biometric parameters under ultrasound imaging. Using a high-frequency ultrasound system with a 40 MHz transducer, we measured 293 fetuses of biparietal diameter and mean abdominal diameter from day 12.5 postcoitus (p.c.) until day 18.5 p.c every day. Thirteen algorithms for humans based on head and/or abdominal measurements were assessed. We established an accurate formula based on measurement of the abdomen in Jcl:ICR mice to investigate gestational complications, such as intrauterine growth restriction.

Elevated level of plasma vascular endothelial growth factor after gonadotropin-releasing hormone agonist treatment for leiomyomata

Objective. Uterine leiomyomata are the most common gynecological benign tumor and greatly affect reproductive health and well-being. The pathophysiology and epidemiology of fibroids are poorly understood. Gonadotropin-releasing hormone agonist (GnRH-a) pretreatment is one of the unfavourable factors for leiomyomata treatment with uterine artery embolisation (UAE). In this study, we investigated the plasma level of vascular endothelial growth factor (VEGF) in uterine leiomyoma patients with or without GnRH-a pretreatment. Study design. Thirty-two women who underwent UAE for symptomatic uterine leiomyoma were analysed. The plasma level of VEGF was studied before UAE. Results. The level of plasma VEGF was significantly higher in the GnRH-a pretreated group compared with the non-treated group. Conclusion. A compensative reaction for vasculature after GnRH-a treatment is speculated. Higher level of VEGF in GnRH-a pretreatment group could be one of the unfavourable factors for the treatment of uterine leiomyomata by UAE.

VEGF Targeting Agents in Ovarian Cancer

Angiogenesis, the formation of new blood vessels, is a critical component in the growth and metastasis of cancers and has been recognized as an attractive target for anticancer therapy (Ferrara, 2002). Among the pro-angiogenic factors, vascular endothelial growth factor (VEGF) is recognized as the predominant mediator of angiogenesis in tumor cells (Ferrara & Kerbel, 2005). As VEGF is overexpressed in most ovarian cancers, the VEGF pathway is a promising target for anti-angiogenic therapy against ovarian cancer (Burger, 2011). Recent increases in our understanding of the molecular pathways that control tumor angiogenesis have led to the development of novel VEGF-targeting agents for the treatment of ovarian cancer (Burger, 2011). In addition to inhibiting neo-vascularization, antiangiogenic agents are also believed to normalize intratumoral blood vessels. Intratumoral vessels are hyperpermeable, leading to interstitial hypertension and impaired perfusion in tumors. Normalization of the tumor vasculature results in a reduction in interstitial pressure and the improved delivery of oxygen, nutrients, and cytotoxic agents (Ferrara, 2002). Many of these agents have been evaluated in clinical trials, and some of them have shown promising clinical activity against ovarian cancer (Burger, 2011). In this article, we review the emerging VEGF-targeting strategies for treating ovarian cancer and provide information about the latest clinical studies of VEGF-targeting agents that have been employed as treatments for ovarian cancer.