KAFKAS UNIVERSITESI VETERINER FAKULTESI DERGISI

In this study, to determine the mechanisms of cell death in developing follicles, we investigated whether expression of Bcl-2, p53 and Bax play a role throughout the growth of follicles in the mice. Ovarian tissues and oocytes were obtained from 30 Mus Musculus type mouse. The immunostaining of ovarian tissue sections and oocytes was performed using anti-Bcl-2, anti-Bax and anti-p53 antibodies and terminal deoxynucleotidyl transfrase (TdT) assay (TUNEL) were also used for detection of internucleosomal DNA fragmentation. In ovarian tissue section evaluation, granulosa cells in antrum of secondary and Graaf follicles were positive with TUNEL staining. Immunoreactivity of Bcl-2 was moderate in primary and secondary follicles of granulosa cells. While immunostaining of p53 was moderate in granulosa cells of Graaf follicles, Bax immunoreactivity was moderate and strong in secondary and Graaf follicles, respectively. When granulosa cells were break off from zona pellucida, there were TUNEL positive cells. In unfertilised oocytes evaluation, TUNEL positive cells were observed in the granulosa cells which were free from zona pellucida. When the granulosa cells were attached with zona pellucida, they were TUNEL negative. While immunoreactivity of Bcl-2 was detected in both oocytes and granulosa cells which were attached with zona pellucida, immunostaining of p53 were only detected in granulosa cells which break off from zona pellucida. In conclusion, regulation of apoptosis in granulosa cells may be controlled by Bax expression and when the granulosa cells were not attached with zona pellucida, they may go into the apoptotic cascade. Therefore; we suggest that, the death of granulosa cells may control signals from intrinsic pathways in the Graaf follicule or from extrinsic pathways after ovulation. However, we consider of further studies to be necessary

REPRODUCTIVE SCIENCES

alpha-Lipoic acid (ALA) is a safe natural molecule involved in the immunomodulation of many physiological processes. Orally administered ALA has been reported to treat several inflammatory pathologies and support pregnancy. Our study aimed at testing ALA vaginal administration in female Wistar rats evaluating its tissue distribution (experiment I), impact on implantation process (experiment II), and effectiveness in contrasting induced preterm birth (experiment III). In experiment I, rats were intravaginally treated with 50 mg/kg or 500 mg/kg ALA, or with a physiologic solution, for 4 days. alpha-Lipoic acid distribution in uterus and cervical tissues was evaluated by immunohistochemical analyses. In experiment II, rats received intravaginally the above treatments for 5 days, then they were mated and, if pregnant, included in the experiment to evaluate both implantation rate and the content of implantation mediators in uterus tissues. In experiment III, pregnant rats were pretreated with placebo or with vaginal ALA for 4 days and then induced to delivery with mifepristone plus PGE2 on the 19th day of pregnancy. The delivery time was recorded, and the messenger RNA (mRNA) levels of pro-inflammatory cytokines were detected in the uterine tissues by real-time polymerase chain reaction. Immunohistochemistry was also performed. Results showed that vaginal ALA was well absorbed and distributed. The treatment did not affect the implantation process and was able to significantly revert mifepristone plus prostaglandin E2 effects, delaying the timing of delivery and significantly decreasing mRNA synthesis and release of pro-inflammatory cytokines. We provide for the first time new information on vaginal ALA use, even during pregnancy, opening a perspective for further studies

JOURNAL OF MOLECULAR HISTOLOGY

Aim of this study was to investigate the effects of lipoic acid on uterine wound healing by immunohistochemical and biochemical assay in a rat uterine horn model with full thickness injury. Thirty-two female Wistar albino rats were randomised into five groups: Control group, with no intervention; uterine scar group 15days (US15d), uterine scar group 15 days + alpha lipoic acid (ALA) (US15d + ALA), uterine scar group 30 days (US30d) and uterine scar group 30 days + ALA (US30 days + ALA). After uterine incision 100 mg/kg of ALA was administered by oral gavage for either 15 or 30 days. Vascular endothelial growth factor (VEGF) and alpha smooth muscle actin (alpha-SMA) distribution were evaluated by immunohistochemical methods in tissue and ELISA methods in tissue homogenate. The percentage of alpha-SMA positive area in US15d + ALA and US30d + ALA groups was significantly higher than US15 and US30d groups. The percentage of VEGF positive area in US15d + ALA group was significantly higher than US15d group and US30d + ALA group was significantly higher than US30d group. Biochemically, alpha-SMA was significantly higher in the US15d + ALA group when compared to US15d group and higher in US30d + ALA group when compared to US30d group. VEGF was significantly higher in US15d + ALA and US30d + ALA groups when compared to US15 and US30d groups. In conclusion, ALA was found to be effective in enhancing wound healing in uterine full thickness injury

α-Lipoic Acid Vaginal Administration Contrasts Inflammation and Preterm Delivery in Rats

alpha-Lipoic acid (ALA) is a safe natural molecule involved in the immunomodulation of many physiological processes. Orally administered ALA has been reported to treat several inflammatory pathologies and support pregnancy. Our study aimed at testing ALA vaginal administration in female Wistar rats evaluating its tissue distribution (experiment I), impact on implantation process (experiment II), and effectiveness in contrasting induced preterm birth (experiment III). In experiment I, rats were intravaginally treated with 50 mg/kg or 500 mg/kg ALA, or with a physiologic solution, for 4 days. alpha-Lipoic acid distribution in uterus and cervical tissues was evaluated by immunohistochemical analyses. In experiment II, rats received intravaginally the above treatments for 5 days, then they were mated and, if pregnant, included in the experiment to evaluate both implantation rate and the content of implantation mediators in uterus tissues. In experiment III, pregnant rats were pretreated with placebo or with vaginal ALA for 4 days and then induced to delivery with mifepristone plus PGE2 on the 19th day of pregnancy. The delivery time was recorded, and the messenger RNA (mRNA) levels of pro-inflammatory cytokines were detected in the uterine tissues by real-time polymerase chain reaction. Immunohistochemistry was also performed. Results showed that vaginal ALA was well absorbed and distributed. The treatment did not affect the implantation process and was able to significantly revert mifepristone plus prostaglandin E2 effects, delaying the timing of delivery and significantly decreasing mRNA synthesis and release of pro-inflammatory cytokines. We provide for the first time new information on vaginal ALA use, even during pregnancy, opening a perspective for further studies