Abnormal Uterine Bleeding Is Associated With Increased BMP7 Expression in Human Endometrium.

Abnormal uterine bleeding (AUB), a common health concern of women, is a heterogeneous clinical entity that is traditionally categorized into organic and nonorganic causes. Despite varied pharmacologic treatments, few offer sustained efficacy, as most are empiric, unfocused, and do not directly address underlying dysregulated molecular mechanisms. Characterization of such molecular derangements affords the opportunity to develop and use novel, more successful treatments for AUB. Given its implication in other organ systems, we hypothesized that bone morphogenetic protein (BMP) expression is altered in patients with AUB and hence comprehensively investigated dysregulation of BMP signaling pathways by systematically screening 489 samples from 365 patients for differences in the expression of BMP2, 4, 6, and 7 ligands, BMPR1A and B receptors, and downstream SMAD4, 6, and 7 proteins. Expression analysis was correlated clinically with data abstracted from medical records, including bleeding history, age at procedure, ethnicity, body mass index, hormone treatment, and histological diagnosis of fibroids, polyps, adenomyosis, hyperplasia, and cancer. Expression of BMP7 ligand was significantly increased in patients with AUB (H-score: 18.0 vs 26.7; P < .0001). Patients reporting heavy menstrual bleeding (menorrhagia) as their specific AUB pattern demonstrated significantly higher BMP7 expression. Significantly, no differences in the expression of any other BMP ligands, receptors, or SMAD proteins were observed in this large patient cohort. However, expression of BMPR1A, BMPR1B, and SMAD4 was significantly decreased in cancer compared to benign samples. Our study demonstrates that BMP7 is a promising target for future investigation and pharmacologic treatment of AUB.

Analysis of the reason of abnormal uterine bleeding induced by copper corrosion of IUD Cu.

OBJECTIVES: This study aimed to analyze relationship of the copper corrosion of copper intrauterine device (TCu220 IUD) and abnormal uterine bleeding. MATERIALS AND METHODS: Sixty-four patients of abnormal uterine bleeding (too much blood volume, shorten cycle, lengthen period or irregular vaginal bleeding) and 72 cases of normal menstrual cycle and quantity in the present hospital, which were removed of IUD due to non-medical reasons, were enrolled, and 36 regular menstruation cases without placing IUD were selected as control group, in which had assessed in vitro copper ion release of TCu220 IUD and content of copper ions and VEGF in endometrial tissue of each group of women. RESULTS: Daily Cu I UD copper dissolution quantities of abnormal uterine bleeding women was significantly higher than that of regular menstruation women (p < 0.05). Copper ion content and the expression of VEGF in endometrial tissue of abnormal uterine bleeding women was significantly higher than that of regular menstruation women endometrial tissue (p < 0.05), and the endometrial VEGF expression had a positive correlation with copper ion concentration in endometrial tissue. CONCLUSION: High dissolution quantity of Cu IUD may lead to increase of copper ion content in endometrial tissue and may cause VEGF secretion in the endometrium, and then the occurrence of abnormal uterine bleeding.

Estrogen up-regulates MMP2/9 expression in endometrial epithelial cell via VEGF-ERK1/2 pathway.

OBJECTIVE: To study the effect of estrogen on anovulatory dysfunctional uterine bleeding (ADUB). METHODS: Primary endometrial epithelial cells of Hainan Lizu female was cultured and hydrolytic activity of gelatinase was determined by gelatin zymography analysis. Cellular mRNA and protein synthesis was blocked respectively to determine whether the increased expression of MMP-2/9 was induced by estrogen. The expression of VEGF was blocked by siRNA. After treatment with various factors, MMP-9, VEGF, total Erk and phosphorylated Erk expression in primary uterine epithelial cells was detected by Western blotting analysis. Cell MMP-2/9mRNA levels was measured by real-time RT-PCR. RESULTS: The activity and expression of MMP2/9 was increased in the endometrium of patients with ADUB. Estrogen could up-regulate the expression of VEGF and activate Erk 1/2-Elk1 signal path. After interference by siRNA, ERK1/2 pathway was blocked in cells, and the expression of MMP-2/9 was down-regulated. ERK1/2 specific blocker U0126 blocked ERK phosphorylation, and it could down-regulate the expression of MMP-2/9. CONCLUSIONS: The results showed that the estrogen can increase the expression of VEGF, and thus activate ERK1/2 pathway to induce MMP-2/9 expression.

Iatrogenic unscheduled (breakthrough) endometrial bleeding.

Erratic, "unscheduled", vaginal bleeding continues to be the greatest disadvantage associated with widespread use of long-acting, progestogen-only methods of contraception. As a consequence, it is also the main reason for premature discontinuation of use of these methods in most cultures. From other perspectives, these methods have high acceptability, very high contraceptive efficacy and a range of valuable, added, non-contraceptive health benefits. There has been widespread awareness of the variability of these vaginal bleeding patterns associated with long-acting methods for several decades and much research has been invested into studying their patterns and implications. Considerable research has also been directed towards trying to understand the underlying mechanisms responsible for the unpredictable bleeding. Much has been clarified about the multiple mechanisms contributing to the appearance of superficial, thin-walled fragile vessels within the endometrium of many of those women with troublesome bleeding, but there is still little understanding of why some women develop these vessels and others have no fragile vessels (and may therefore develop amenorrhea). We now have several medical approaches to reliably stopping a prolonged episode of troublesome bleeding, but no good therapy to produce long-lasting relief from recurrence of erratic bleeding in predisposed women. Future understanding of the variability in individual endometrial responses in different women may be a key to solving this frustrating symptom.

[Clinical analysis of 106 cases on effect of supplementing qi and nourishing yin recipe in treating anovulatory dysfunctional uterine bleeding].

OBJECTIVE: To observe the clinical treatment of supplementing qi and nourishing yin recipe and progestogen. METHOD: One group was treated using traditional Chinese medicine decoction, which was made of Danshen, Shenghuangqi, Wuweizi, Baishao, Nvzhenzi, Hanliancao and Wuzeigu, and the other group was medroxyprogesterone acetate (depogeston). RESULT: After 3 menstrual cycles of treatment, the recovery rate of the treatment group was 43.42%, with 27.63% of marked effectiveness,18.42% of positive effects, and 10.53% of inefficiency. The total effective rate was 89.47%. The recovery rate of the control group was 23%, with 47% of marked effectiveness, 20% of positive effects, and 10% of inefficiency. The total effective rate was 90.00%. With statistically comparison, there was no obvious difference. The treatment group was higher than the control group on the Chinese symptom, the two groups were significantly different (P < 0.05). The treatment group and the control group was no obvious difference on Hb, PLT, PT, APTT. The treatment group and the control group was no obvious difference on FSH, LH, E2, P. CONCLUSION: Traditional Chinese medicine decoction should be recommended for its better therapeutic efficacy and stable clinical effects such as high recovery rate, significant improvement of symptom and no side effect and no adverse reaction.

Enhanced expressions of matrix metalloproteinase (MMP)-2 and -9 and vascular endothelial growth factors (VEGF) and increased microvascular density in the endometrial hyperplasia of women with anovulatory dysfunctional uterine bleeding.

OBJECTIVE: To evaluate the roles of endometrial matrix metalloproteinase (MMP)-2 and -9, vascular endothelial growth factors (VEGF), and microvascular density (MVD) in the occurrence of anovulatory dysfunctional uterine bleeding (DUB). DESIGN: A prospective analytical design. SETTING: The obstetrics and gynecology department of an academic training hospital. PATIENT(S): Sixty women with anovulatory DUB and 20 control women. INTERVENTION(S): Endometrial biopsies were obtained for the assessment of immunohistochemical staining of MMP-2 and -9, VEGF, and endometrial MVD using an antibody to CD34. MAIN OUTCOME MEASURE(S): The results were determined through the expressions of MMP-2 and -9, VEGF, and CD34. RESULT(S): The frequencies of MMP-2 and -9 expression in endometrial stroma and of VEGF expression in endometrial glands were all significantly higher in the endometrial hyperplasia of women with anovulatory DUB than they were in the control group. Additionally, the mean score of endometrial MVD was significantly higher in the endometrial hyperplasia of women with anovulatory DUB than it was in the control group. In women with anovulatory DUB, VEGF expression in endometrial glands was statistically correlated with MMP-2 and -9 expressions in endometrial stroma and endometrial MVD. CONCLUSION(S): These results suggest that enhanced expressions of MMP-2 and -9, VEGF, and increased MVD in endometrial hyperplasia may play important roles in the pathogenesis of women with anovulatory DUB.

Activation of NF-kappaB and COX-2 expression is associated with breakthrough bleeding in patients using oral contraceptives in extended regimens.

The objective of the present study was to determine whether there is an increase in endometrial inflammation associated with the occurrence of breakthrough bleeding in patients using an oral contraceptive in extended regimens. The presence of nuclear factor NF-kappaB and Cox-2 expression was determined by immunohistochemistry in endometrial samples removed by hysteroscopy from patients with breakthrough bleeding during continuous use of an oral contraceptive containing gestodene. All patients had a history of menorrhagia associated or not with the presence of uterine pathology. The percentage of endometria showing a positive staining reaction for NF-kappaB in cell nuclei was significantly higher in patients with breakthrough bleeding than in those with amenorrhea. Cox-2 expression in the endometrium was also significantly more frequent in patients with breakthrough bleeding. The occurrence of breakthrough bleeding in patients with uterine pathology using combined oral contraceptives is associated with the activation of endometrial inflammation through the NF-kappaB pathway.

Effects of doxycycline on serum and endometrial levels of MMP-2, MMP-9 and TIMP-1 in women using a levonorgestrel-releasing subcutaneous implant.

BACKGROUND: Endometrial spotting and/or bleeding (ESB) occurs in levonorgestrel subcutaneous implant (LNG SI) users. Matrix metalloproteinases (MMPs) may play a role in ESB. STUDY DESIGN: Women between 18 and 40 years with regular menstrual cycles had a baseline evaluation followed by LNG SI insertion and randomization to doxycycline (DOX; 20 mg) or placebo (PL) twice a day. MMP-2, MMP-9 and tissue inhibitor of MMP-1 (TIMP-1) in serum and the endometrium were estimated at baseline and at 1, 3 and 6 months after insertion. RESULTS: LNG increased serum MMP-9, while DOX decreased MMP-9 levels compared to PL after 1 month (p<.05). DOX decreased endometrial MMP-9 at 1 and 6 months compared to baseline and PL (p<.05). DOX increased endometrial TIMP-1 at 6 months compared with baseline and PL (p<.05). MMP-2 levels were unchanged. CONCLUSION: LNG SI increased serum MMP-9 and TIMP-1 levels, while DOX decreased both serum and endometrial MMP-9 levels.

Calretinin and CD34 immunoreactivity of the endometrial stroma in normal endometrium and change of the immunoreactivity in dysfunctional uterine bleeding with evidence of 'disordered endometrial stroma'.

AIMS: We investigated the pattern of reactivity of calretinin and CD34 in normal and pathological endometria. METHODS: Various endometrial tissues were submitted for calretinin and CD34 immunostaining. RESULTS: Calretinin reactivity was limited to the endometrial stromal cells (ESC) of the superficial zone of the functionalis layer (FL) in the proliferative phase, and was extensive in all stages of the secretory phase. The ESC of the post-menopausal, ectopic, hyperplastic or neoplastic endometria showed negative or focal weak reactivity for calretinin. In dysfunctional uterine bleeding (DUB) with a normal or an abnormal histopathological appearance on routine stain, there were varying degrees of focal to extensive decreases in calretinin reactivity. The foci of negative calretinin reactivity in the FL displayed varying reactivity for CD34 and appeared to be continuous with the basalis layer (BL). Endometrial polyps were often reactive for CD34, but not reactive for calretinin. CONCLUSIONS: Immunostaining for calretinin and CD34 is helpful in the diagnosis of endometrial polyp and hyperplasia. In DUB, with or without abnormal histopathological findings, there were alterations of the zonal pattern of calretinin reactivity in the FL. This alteration appears to be an expansion of the stroma of the BL into the FL, resulting in a 'disordered endometrial stroma'.

Production of interleukin-1alpha by human endometrial stromal cells is triggered during menses and dysfunctional bleeding and is induced in culture by epithelial interleukin-1alpha released upon ovarian steroids withdrawal.

CONTEXT: Endometrial breakdown during menstruation and dysfunctional bleeding is triggered by the abrupt expression of matrix metalloproteinases (MMPs), including interstitial collagenase (MMP-1). The paracrine induction of MMP-1 in stromal cells via epithelium-derived IL-1alpha is repressed by ovarian steroids. However, the control by estradiol (E) and progesterone (P) of endometrial IL-1alpha expression and bioactivity remains unknown. OBJECTIVE AND DESIGN: Variations of endometrial IL-1alpha mRNA and protein along the menstrual cycle and during dysfunctional bleeding were determined using RT-PCR, in situ hybridization, and immunolabeling. The mechanism of EP control was analyzed using culture of explants, laser capture microdissection, and purified cells. Data were compared with expression changes of IL-1beta and IL-1 receptor antagonist. RESULTS: IL-1alpha is synthesized by epithelial cells throughout the cycle but E and/or P prevents its release. In contrast, endometrial stromal cells produce IL-1alpha only at menses and during irregular bleeding in areas of tissue breakdown. Stromal expression of IL-1alpha, like that of MMP-1, is repressed by P (alone or with E) but triggered by epithelium-derived IL-1alpha released upon EP withdrawal. CONCLUSIONS: Our experiments in cultured endometrium suggest that IL-1alpha released by epithelial cells triggers the production of IL-1alpha by stromal cells in a paracrine amplification loop to induce MMP-1 expression during menstruation and dysfunctional bleeding. All three steps of this amplification cascade are repressed by EP.

[The expression of apoptosis related genes Bcl-2 and Bax protein in dysfunctional uterine bleeding].

OBJECTIVE: To research the expression of apoptosis related genes Bcl-2 and Bax protein in dysfunctional uterine bleeding. METHODS: Detecting the expression of Bcl-2 and Bax protein in 40 cases of endometrium tissue of dysfunctional uterine bleeding patients with immunohistochemistry antibiotics protein-peroxide emzyme (SP)methods. RESULTS: (1) The expression of Bcl-2 protein changes clearly and periodically in endometrium in normal cycle, the difference is obvious (P < 0.05). (2) The Bcl-2 protein develops with the hyperplasia in endometrium and the expression intensity is enhanced, the difference is obvious( P < 0.05). (3) The expression of Bax gene is masculine in endometrium in normal menstrual cycle. (4) The expression of Bax gene descends gradually with hyperplasia in uterine endometrium (P < 0.05). CONCLUSION: Apoptosis of endometrium cell is restrained from the excessive expression of Bcl-2 protein and deficient expression of Bax protein, so the uterine endometrium gets to hyperplasia or notypical hyperplasia.

The long-term actions of etonogestrel and levonorgestrel on decidualized and non-decidualized endometrium in a mouse model mimic some effects of progestogen-only contraceptives in women.

Breakthrough bleeding (BTB), a major side effect of long-acting progestogen (p)-only contraceptives in women, is the main reason for discontinuation of their use. To understand the mechanisms of BTB, a mouse model of endometrial breakdown and repair was adapted to evaluate the effects of long-term progestogens on the endometrium. Appropriately prepared mice received either an etonogestrel (ENG)- or levonorgestrel (LNG)-releasing subdermal implant. Forty eight hours after decidualization was induced in one uterine horn the majority of tissues were highly decidualized, designated 0 day (0d). Uteri were collected subsequently at 5-day intervals (to 45d) and both decidualized and non-decidualized horns were analysed for morphological changes, leukocyte infiltration and matrix metalloproteinase expression (MMP). In decidualized horns, large blood vessels (BV) developed and disturbance of tissue integrity was observed at 5d with substantial stromal breakdown by 10d, progressing until 25d when re-epithelialization was initiated. By 45d, the tissue was restored to its pre-decidualized state but with considerable tortuosity of the luminal epithelium. Tissue remodelling was not apparent in the non-decidualized horns before 35d, when hyperproliferation of the luminal epithelium resulted in tortuosity. Changes in morphology were similar with the two progestogens, but occurred more rapidly with LNG. Apart from macrophages, few leukocytes were present in non-decidualized horns but large infiltrates of neutrophils and uterine natural killer cells (uNK) were associated with tissue breakdown in decidualized tissue, many of these cells were MMP9-positive. MMP7 was primarily associated with tissue repair. Therefore, this model mimics some of the changes observed in endometria of women using p-only contraceptives and provides an opportunity for functional studies.

Calretinin: an immunohistochemical marker for the normal functional endometrial stroma and alterations of the immunoreactivity in dysfunctional uterine bleeding.

Calretinin has been identified in the central nervous system, in various endocrine and mesothelial cells, and is often used as an immunohistochemical tool in the pathologic diagnosis. We have recently observed its presence in the endometrial stromal cells (ESC) of the normal functionalis (FL). Endometrial tissue from various physiologic and pathologic conditions was submitted for immunostaining for calretinin. For each condition, two to ten samples were tested. Calretinin displayed a strong cytoplasmic and occasionally nuclear reactivity for ESC of the normal FL in all physiologic phases of the normal menstrual cycle except for the breakdown period. Reactivity was limited in the superficial zone of the FL in proliferative phase, and was extensive in all stages of secretory phase. The ESC of the basilis layer, postmenopausal women, endometriotic and adenomyotic tissue, endometrial polyps, hyperplasia, carcinoma, and ESC neoplasms were not reactive for the marker. In endometrial specimens from patients with dysfunctional bleeding, there were varying degrees of decrease in reactivity in a patchy pattern. We demonstrated for the first time that calretinin reactivity of the ESC is strong and diffuse in a zonal pattern in the normal FL of the normal cycling endometrium. In endometrium with dysfunctional bleeding, postmenopausal, ectopic, hyperplastic, and neoplastic endometria, the ESC displayed focally decreased to negative reactivity. Calretinin can be used as a marker for normally functional ESC. Further study is necessary to investigate the mechanism and the role that calretinin plays in the physiologic cyclic changes of the endometrium.

Apoptosis in endometria of dysfuntional uterine bleeding women.

The study of apoptosis in endometrium of women with irregular uterine bleeding and its predictive value in endometrial malignancy. Analyze apoptotic and mitotic indices and their relevance in irregular uterine bleeding. To determine the expression of Bcl-2 oncoprotein in endometrial glands from patients with irregular uterine bleeding. Department of pathology in a Government Hospital serving a varied socio-economic population in Chennai. Random samples of endometrial currettings from dysfunctional uterine bleeding (DUB) patient who underwent endometrial curettage as therapeutic and diagnostic procedure during the year 2000. Of 50 cases of endometrial samples from patients diagnosed as cases of DUB, the apoptotic and mitotic indexing was carried out and histological categorization revealed 13 cases as Anovulatory. 14 as simple hyperplasia, 5 as early secretory endometrium, 4 as mid secretory and 4 as late secretory endometrium and 7 as endometrium showing features of hormonal imbalance. Three cases were not included, due to sub-optimal processing. A good correlation of the Bcl-2 expression and the apoptotic cell morphology/indices, in the different categories of the endometria of DUB cases is observed. This preliminary study gives an insight to the existence of a correlative pattern of apoptosis in DUB cases. A prospective study on a larger number of cases may substantiate the hypothesis that the Apoptotic and Mitotic indices are useful screening methods with predictive values on development of endometrial carcinoma. It is observed that an increased apoptotic index correlating with high Bcl-2 expression, reflecting the actual cell burden. This prolonged cell survival resisting cell deletion is associated with irregular uterine bleeding endometria.

Glycogen metabolism of the human endometrium.

The glycogen concentration and the activities of glycogen synthetase and phosphorylase were assayed in endometrial tissue obtained during curettage. Tissues were obtained during different phases of the menstrual cycle from women investigated because of menometrorrhagia or sterility. The glycogen concentration was less than 2 mg/g wet wt in specimens from days 1-15 and increased 10-fold during the early secretory phase. The total activities of glycogen synthetase and phosphorylase were increased 1.5- and 2-fold, respectively, during the secretory phase compared to those during the first half of the cycle. The activities of the active forms (a form) of these enzymes did not show significant changes during the menstrual cycle. The largest difference between the proliferative and secretory phases was in the activity of glycogen synthetase phosphatase, which was virtually inactive in tissues obtained on days 1-15 and increased almost 20-fold during the secretory part of the cycle. In tissues obtained from cases of sterility, the activities of glycogen synthetase and phosphorylase were significantly reduced only during the early secretory phase. Glycogen concentration throughout the menstrual cycle and enzyme activities during the midsecretory and premenstrual phase were similar in cases of sterility and other pathologies.