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    A rat study on the PTEN expression in ovarian tissue in PCOS and folliculogenesis
    (Nature Portfolio, 2023) Namli Kalem, Muberra; Anadol, Elvan; Kalem, Ziya; Sezginer, Perihan Yalcinkaya; Elmas, Cigdem; Yilmaz, Canan; Bakirarar, Batuhan
    The objective of this investigation was to examine alterations in PTEN expression within ovarian tissue in a rat model of polycystic ovary syndrome (PCOS). The analysis also encompassed the examination of PTEN alterations in the ovarian tissue throughout the process of folliculogenesis in rats with normal ovulatory cycles. The study involved 12 adult female Sprague-Dawley rats randomly assigned to the letrozole-induced polycystic ovary syndrome (PCOS) group as part of an animal-based research endeavour. The sections derived from the ovaries were subjected to immunohistochemical staining for PTEN. The evaluation of PTEN staining levels in ovarian tissues was conducted using electron microscopy. Follicle counts, as well as hormonal and biochemical analyses (serum luteinising hormone (LH), follicle-stimulating hormone (FSH), anti-Mullerian hormone (AMH), testosterone, oestradiol levels and serum glucose, triglyceride, HDL and LDL-cholesterol levels), were conducted to provide evidence of the manifestation of polycystic ovary syndrome (PCOS) in rats. The number of primordial and Graafian follicles in the PCOS group decreased significantly, and the number of primary, secondary and antral follicles increased significantly. PTEN expression was found to be significantly higher in the PCOS group than in the control group in the primordial follicle oocyte cytoplasm, primordial follicle granulosa cells, primary follicle oocyte cytoplasm, primary follicle granulosa cells, antral follicle oocyte cytoplasm, antral follicle granulosa cells, and corpus luteum (p = 0.007, p = 0.001, p = 0.001, p = 0.001, p = 0.001, p = 0.002, and p = 0.018, respectively). In the non-PCOS group, a time-dependent comparison of the amount of oocyte cytoplasm and PTEN staining in granulosa cells of the oocytes at different stages of development was performed. While the follicles were developing from the primordial follicle to the primary and antral follicle, the amount of PTEN staining in the oocyte cytoplasm decreased, whereas the PTEN activity in the granulosa cells increased as the oocyte developed (p = 0.001 and p = 0.001, respectively). The current investigation demonstrated changes in PTEN expression in ovarian tissue throughout the course of normal folliculogenesis, as well as in instances of disrupted folliculogenesis, with a focus on rats with PCOS.
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    Performance of oral Bosentan-loaded SNEDDS and S-SNEDDS tablets: Biodistribution in mice, echocardiography and histology studies in pulmonary arterial hypertension rat model
    (Elsevier, 2025) Usta, Duygu Yilmaz; Olgac, Seval; Demirel, Murside Ayse; Kula, Serdar; Elmas, Cigdem; Sezginer, Perihan; Kavgaci, Akif
    Bosentan monohydrate (BOS) is the most preferred molecule for treating the rare pulmonary arterial hypertension (PAH) disease. BOS shows low solubility and high variability when administered orally. This study evaluated the pharmacodynamic biodistribution, echocardiography, and histology results of BOS-loaded SNEDDS and BOS-loaded S-SNEDDS tablets. Pharmacodynamic biodistribution studies were conducted with male Balb/c mice (8 weeks old, 18-20 g) after oral administration. XenoLight (TM) DiR and VivoTag (R) 680XL fluorescent dyes were used to monitor biological distribution and absorption with the In Vivo Imaging System (R) (IVIS (R)). Pharmacodynamic echocardiography and histology studies were carried out with Wistar rats (8-10 weeks old, 250-300 g). The PAH rat model was successfully induced with monocrotaline (MCT), which is one dose (60 mg/kg) was intraperitoneally injected. The reference drug (Tracleer (R) 125 mg tablet) and BOS-loaded SNEDDS and S-SNEDDS tablets were administered as 50 mg/kg; 2 mL per os to the treatment groups. Pharmacodynamic biodistribution studies showed that real-time biodistribution in the body, ex-vivo region of interest (ROI) values of organs, and total fluorescence emission were increased (p < 0.05). It has been confirmed that the formulations enter the systemic circulation via the lymphatic system, do not have a first-pass effect in the liver, and show no emission in the liver. The echocardiographic study was performed for up to 14 days and no difference was found between the treatment groups which are the reference tablet (Tracleer (R)), BOS-loaded SNEDDS, and BOS-loaded S-SNEDDS tablet (p > 0.05). Hematoxylin and Eosin (H&E) and Immunohistochemical (IHC) staining were done for the histology studies. These studies showed that the BOS-loaded formulations have a similar therapeutic effect on histopathological phenomena in lung and liver tissues. As the histological evaluation results, lower-dose formulations were found to be more effective than the same dose reference tablet in terms of improvements in histological parameters (p < 0.05). Comprehensive and comparative in vitro and in vivo studies indicate that BOS-loaded formulations could be an alternative oral drug delivery system for PAH treatment compared to the reference product.
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    The effect of controlled ovarian hyperstimulation on ovarian reserve via PTEN pathway
    (Bioscientifica Ltd, 2022) Sezginer, Perihan; Elmas, Cigdem; Yildiz, Fatma
    This study was carried out to investigate whether repeated controlled ovarian hyperstimulation (COH) affects ovarian reserve. For this reason, we aimed to show possible changes in the expression of PTEN and FOXO3, which are involved in preserving the over-reserve, after applying the COH protocol methods. For this purpose, 18 young Wistar albino female rats (8 weeks old) were randomly assigned as group 1 (control), group 2, and group 3 as 6 subjects in each group. Experimental groups were treated with 10 IU/0.1 mL pregnant mare's serum gonadotropin and a COH protocol consisting of 10 IU/0.1 mL human chorionic gonadotropin injection after 48 h. This procedure was applied three and five times to group 2 and group 3, respectively. For the control groups, the same procedures were performed with 0.1 mL of 0.9% sodium chloride solution. At the end of the experiment, the ovarium tissues were placed in a 10% neutral formaldehyde solution for light microscopic examinations. In histological sections stained with hematoxylin and eosin, the number of ovarian follicles was determined using the physical dissector method. However, the expression of PTEN, FOXO3, and LH-R molecules was evaluated by immunohistochemical methods. As a result of our study, it was concluded that COH administration reduces the expression levels of PTEN and FOXO3 proteins and LH-R, which are among the essential components of the PIK3 intracellular signaling pathway and also increased the levels of hormones such as follicle-stimulating hormone, estradiol, and luteinizing hormone, which are over-reserve markers, and causes adverse effects on the histological structure, oocyte morphology, and number of ovaries. Lay summary Today, approximately 10-15% of couples experience fertility problems. However, assisted reproductive techniques help people with fertility problems to get pregnant. The main purpose of these techniques is to put the sperm and egg together outside the woman's body where the eggs are fertilized and then to return the fertilized eggs (embryos) to the womb. During a woman's menstrual cycle, several hormones influence the growth of the eggs. This process can be mimicked by using various medications. Medication is given to increase the number of eggs that develop. However, this method is not the same as normal ovulation. Therefore, in our study, we wanted to examine the effect that developing multiple follicles has on the number and quality of eggs remaining for the future.