Evaluation of changes in the expression of Wnt/beta-catenin target genes in mouse reproductive tissues during estrous cycle: an experimental study

Background: The Wingless-type [Wnt]/beta-catenin signaling pathway controls cell homeostasis. Reproductive tissues are dynamic in response to steroidal hormone changes. Steroidal hormones are known to control the Wnt/beta-catenin pathway, but their role in reproductive tissues remains unknown

Objective: The present study aims to investigate the expression patterns of Wnt/beta-catenin target genes in mouse reproductive tissues during the normal estrous cycle

Materials and Methods: In this experimental study, 16 adult NMRI mice were grouped as proestrus, estrus, metestrus, and diestrus according to vaginal smear and histological evaluation of uterine and ovarian tissues. Uterine horns and ovarian tissues were collected. Reverse transcription quantitative polymerase chain reaction was performed to evaluate the expression of Wnt/beta-catenin target genes [Myc2, Ppard, Id2, Birc5, and Ascl2] at different stages of the estrous cycle

Results: The expression levels of Id2, Ascl2, and Pprd in uterine tissue were significantly higher at the proestrus phase than at the other stages. Meanwhile, Birc5 expression in uterine tissue was significantly higher at the metestrus stage than at the other stages. Furthermore, Myc2 expression was significantly higher at the diestrus stage than at the estrus and metestrus stages. In the ovarian tissue, the highest expression of Id2, Ascl2, and Birc5 was detected at the proestrus stage, whereas the highest expression of Myc2 and Ppard was observed at the estrus stage

Conclusion: This study showed that Wnt/beta-catenin target genes profiles are different among estrous cycle. It seems that different hormonal profiles during estrous cycles play a key role in the expression pattern of Wnt/beta-catenin target genes in ovarian and uterine tissue

Antioxidant Activity of Oral Administration of Rosmarinus Officinalis Leaves Extract on Rat's Hippocampus which Exposed to 6-Hydroxydopamine

Carnosic acid, a diterpene of Rosemarinus officinalis leaves extract (RE), has potent antioxidant activity in vitro. The dopaminergic connection of substantia nigra pars compacta to the hippocampus might be affected by oxidative stress which caused cognitive impairment observed in the early phase of Parkinson's disease (PD). Adult male Wistar rats were lesioned bilaterally by intra-nigral injection of 6-OHDA, and divided into six groups: four groups that orally given RE containing 40% of carnosic acid, at doses of 25, 50 and 100 mg/kg (treated rats) and distilled water (H2O), once daily for a period of 14 days before and after the injury. There were also two another groups as control rats which injected by normal saline and untreated lesion group. The injured animals were evaluated for their spatial memory performance by Morris Water Maze test. Lesioned rats showed significant increase in escape latency, as compared with control group. Two weeks after injury, tissue samples were collected from the hippocampus. Levels of catalase (CAT), glutathione peroxidase (GPX) and superoxide dismutase (SOD), malondialdehyde (MDA) and reactive oxygen species (ROS) were determined. There were significant increase of SOD, GPX and CAT enzymes activities in RE50 treated group as compared to lesioned rats. We found a significant decrease of ROS in RE50 treated group as compared to Lesioned rats. These findings provide evidence that 50 mg/kg of RE decreased oxidative damage of the hippocampus induced by 6-OHDA and serve as potential candidate for the treatment of PD.

Effect of wound dressing with fresh kiwifruit on healing of cutaneous wound in rats

The kiwi fruit is known to have dramatic antibacterial, debridement, wound contracture, and angiogenic effects. We propose that kiwifruit is an ideal candidate to enhance the process of wound healing. The present study assessed the effects of wound kiwifruit dressing on cutaneous wound healing in rat. In this experimental study, 30 male Wistar rats were randomly divided into 2 groups of control and kiwifruit group. A full-thickness dermal incision [35 mm length] was made on the right side of the paravertebral region. In the control group, one day after wound induction wounds was dressed with vaseline sterile gauze after normal saline irrigation. In the second group, the wounds were dressed with kiwifruit. Wound healing was evaluated by measuring surface area, percentage of healing, duration of healing, and wound tensile strength. Obtained results showed that the duration of wound healing in kiwi group in comparison with the control was significantly decreased. The amount of wound healing in percent was also significantly different between control and kiwi groups at days 3, 6 [p<0.001], 9 [p<0.05], 12 and 14 [p<0.01]. Comparisons of wound length between control and kiwi group per day showed that kiwi group had significantly lower wound length on day 9, 12, 14 and 16 [p<0.01, 0.001, 0.01 and 0.01, respectively] in comparisons to control group. Also, the wound tensile strength in kiwi group also was significantly greater than the control animals [p<0.01]. We concluded that our study provides some evidence to support the use of kiwi to accelerate wound healing

impact of alpha lipoic acid on developmental competence of mouse vitrified pre-antral follicles in comparison to those isolated from vitrified ovaries

Cryopreservation of ovarian tissues and pre-antral follicles is a promising prospect for preservation of women fertility. The aim of this study was to evaluate the in vitro developmental competence of mouse vitrified pre-antral follicles in comparison to isolated pre-antral follicles derived from vitrified ovaries in the presence of alpha lipoic acid [ALA]. Pre-antral follicles derived from fresh, vitrified-warmed ovarian tissues and vitrified-warmed pre-antral follicles were cultured individually with or without ALA, followed by adding hCG to induce ovulation. The follicle growth, oocyte maturation, and embryo development were assessed. The diameter and development of follicles, oocyte maturation and embryo development rates were significantly higher in ALA supplemented groups compared to the respective ALA-free conditions groups. Aforementioned parameters were significantly higher in vitrified-warmed follicles in comparison to follicles derived from vitrified-warmed ovaries. These findings support a superior performance of pre-antral follicles when vitrified rather than when isolated from vitrified ovaries with regard to increasing the rates of developmental parameters. Moreover, ALA improves the in vitro maturation of pre-antral follicles in vitrified and non-vitrified samples.

Total oxidative status of mouse vitrified pre-antral follicles with pre-treatment of alpha lipoic acid

Cryopreservation of pre-antral follicles is a hopeful technique to preserve female fertility. The aim of the present study was to evaluate reactive oxygen species [ROS] and total antioxidant capacity [TAC] levels of mouse vitrified pre-antral follicles in the presence of alpha lipoic acid [ALA]. Isolated pre-antral follicles [140-150 micro m in diameter] were divided into vitrified-warmed and fresh groups. Each group was subjected to in vitro maturation with or without ALA for 12 days, followed by adding human chronic gonadotropin to induce ovulation. In vitro fertilization was performed to evaluate their developmental competence. In parallel, the amount of ROS and TAC were assessed after 0, 24, 48, 72, and 96 h of culture by 2',7'-dichlorofluorescin assay and ferric reducing/antioxidant power assay, respectively. The respective rates of survival, antrum formation, and metaphase II oocytes were significantly higher in ALA-supplemented groups compared to the groups not treated with ALA. TAC and ROS levels were significantly decreased and increased, respectively during the culture period up to 96 h in the absence of ALA in both vitrified and non-vitrified samples. However, with pretreatment of ALA, TAC levels were increased significantly and remained constant up to 96 h in vitrified-warmed pre-antral follicles, while ROS levels completely returned to the level of starting point after 96 h of culture in the presence of ALA. Pretreatment of ALA positively influences development of pre-antral follicles in vitrified and nonvitrified samples through increasing follicular TAC level and decreasing ROS levels

Spontaneous expression of neurotrophic factors and TH, Nurr1, nestin genes in long-term culture of bone marrow mesenchymal stem cells

It has been reported that rat bone marrow stromal cells [BMSCs] can be spontaneously differentiated into neural-like cells without any supplemental growth factors and/or chemical treatment after long-term culture. This study aims to determine whether, growth factors secreted by MSCs could induce self-differentiation into neural-like cells in a long-term culture. This study consisted of two groups: i. rat BMSCs [passage 5] were cultured in alfa- minimal essential medium [alpha-MEM] and 10% fetal bovine serum [FBS] without the addition of inducer and exchanging medium for three weeks, as the experimental group and ii.rat BMSCs [passage 5] as the control group. Each group was analysed by reverse transcriptase polymerase chain reaction [RT-PCR] to evaluate the expressions of neurotrophic factors and neural marker genes. Statistical analyses were carried out using one-way analysis of variance [ANOVA] and Tukey's multiple comparison with SPSS software [version 16]. P< 0.05 was considered statistically significant. The experimental group [fifth passage of BMSCs] obtained from adult rats spontaneously differentiated into neural precursor cells after long-term culture. Cultured cells expressed tyrosine hydroxylase [TH], Nurr1 and nestin genes. Furthermore, some growing cells in suspension became neurosphere-like. Self-differentiated rat MSCs [SDrMSCs] expressed significantly higher levels of NGF [0.96 +/- 0.16], nestin [0.63 +/- 0.08], and Nurr1 [0.80 +/- 0.10] genes [p<0.05]. In this study, we reported that rMSCs in long-term culture underwent spontaneous transformation to neural precursors without the supplement of growth factors and specific chemicals. Cells expressed neural markers such as: TH, Nurr1, and nestin genes