| Objective1. To investigate the effects of gonadotropin releasing hormone agonist (GnRHa) on apoptosis of cultured myomatous cell and endometrial cell.2. To investigate the relationship between apoptosis and uterine myoma.To evalute the relationship between apoptosis of cultured endometrial cell and abnormal uterine bleeding. To further explore the mechanism of GnRHa in the treatment on uterine myoma and provide new approach and experimental evidences for the clinical treatment of uterine myoma.Materials and methods1. SamplesFrom Jan 2008 to June 2009, uterine myomatous cells(study group A),myometrial cells (control group A)and endometrial cells(study group B) from 36 women(mean age 38.21±2.28, range 28-46) who received myomectomy for uterine myomas with abnormal uterine bleeding and endometrial cells (control group B) from 22 women(mean age 35.23±3.32, range 22-45) who received ovarian cystectomy surgery for benigh ovarian epithelial tumor hospitalized in gynaecologic department were cultured in vitro. Biopsy specimens were obtained from all subjects in the proliferative phase by endometrial curettage before surgery. Endometrial phase was assigned on the basis of the histologic evaluation and subjects with endometrial hyperplasia or endometrial carcinoma were excluded.All the patients were without any other gynecologic diseases, such as endometriosis, and had not received any hormonal medical treatments during the prior 3 months.2. MethodsLevels of apoptosis were examined in myomatous cells, myometrial cells and endometrial cells before and after incubation with GnRH-a (triptorelin). The percentage of apoptotic cells was evaluated using the terminal deoxynucleotidyl transferase-mediated d-UTP nick end labeling (TUNEL) assay and a flow cytometry method was used to evaluate Annexin V levels.3. Statistics analysisData were recorded as x±S. The results were analyzed by one-factor analysis of variance (ANOVA) with SPSS13.0 statistical software.The results were analyzed by LSD-t if they have homogeneity of variance,by Dunnett-t if they have not.Significance was set at P<0.05.Results1. Cells in two groups showed cell shrinkage, floating nucleus concentration, nuclear fragmentation and typical apoptotic bodies.2. The apoptotic ratio of cultured myomatous cells and endometrial cells without GnRHa were lower than those in control groups and the apoptotic ratio increased with the prolongation of time in two guoups(P<0.01).3. After GnRHa addition,each group showed higher apoptotic ratio and a trend towards higher apoptotic ratio linked to advanced concentration.Furthermore, apoptotic ratios in study groups were significantly higher than those in control groups at the same time point and same concentration(P<0.01).Conclusions1. The lower level of apoptotic ratio of uterine myomatous cells caused by unbalance of apoptotic regulation may be associated with the pathogenesis of uterine myoma.2. The abnormal apoptotic ratio of endometrial cells may be associated with the development of abnormal uterine bleeding.3. GnRHa may increase the apoptotic ratio of cultured uterine myomatous cells and myometrial cells by autocrine or paracrine function, and provides the experimental and theoretical evidence for GnRHa clinical application.4. GnRHa stimulated apoptosis in endometrial cells from patients with abnormal uterine bleeding and this could, at least in part, account for the therapeutic action of GnRHa. |
PDF Full Text Request