Effects of astaxanthin on expression of apoptosis and oxidative stress related genes in H2O2 induced oxidative stress BE(2)-C human neuroblastoma cell line.

Antioxidants may affect the apoptosis induced by oxidative stress experimental models. The present study was conducted to investigate the effects of astaxanthin on expression of apoptosis and oxidative stress-related genes in H2O2 induced oxidative stress BE(2)-C human neuroblastoma cell line. This experimental study consisted of six groups including control, H2O2 induced oxidative stress control, 100 mM vitamin C intervention, 25 µM astaxanthin intervention (Ax1), 50 µM astaxanthin intervention (Ax2) and 100 µM astaxanthin intervention (Ax3). Real-time PCR was used to study the expression of BAX, BCL2, Caspase3 (CAS3), P53, peroxisome proliferator-activated receptor γ (PPARγ), superoxide dismutase (SOD), glutathione peroxidase 1 (GPX), catalase (CAT) and nuclear factor erythroid 2-related factor 2 (NRF2). According to the results, among the apoptosis-related genes, CAS3 was down-regulated in groups vitamin C, Ax1 and Ax2 compared with H2O2 group, while P53 was down-regulated only in group vitamin C (P < 0.05). Among the oxidative stress-related genes, GPX was up-regulated in groups Ax1, Ax2 and Ax3 compared with H2O2 group, while all the experimental groups showed up-regulation for CAT and NRF2 (P < 0.05). In conclusion, astaxanthin as a powerful antioxidant could inhibit apoptosis via amelioration of CAS3 gene which might be through amelioration of some antioxidant-related genes.

Effects of zinc on expression of apoptosis-related genes in freezing thawing damage of adipose tissue derived mesenchymal stromal cells.

The present study was performed to investigate the effects of zinc supplementation on freezing thawing damage in adipose tissue-derived mesenchymal stromal cells (MSC) of mice through studying cellular viability and gene expression profile of apoptosis. Slow freezing method was conducted and the samples were treated with zinc doses 0, 2.5, 5, 10, 25, 50 and 100 µM. Viability was increased in groups of 2.5, 10 and 25 µM zinc in comparison to the control group. Gene expression study showed that in the group of 2.5 µM zinc, Fas, Bax and Caspase3 had down regulation. Up regulation of Bcl2 was observed in the groups of 10 and 25 µM zinc. P53 did not have a protecting regulation in the groups of study. The present study showed that doses 2.5-25 µM of zinc had a rather safe toxicity, increased cellular viability, and ameliorated expression of apoptosis-related genes in both intrinsic and extrinsic pathways.

Effect of Selenium on Expression of Apoptosis-Related Genes in Cryomedia of Mice Ovary after Vitrification.

INTRODUCTION: Freezing of ovarian tissue is used for preservation of fertility. The freezing-thawing process is accompanied by oxidative stress and induction of apoptosis. Apoptosis is a complex process that has been studied in animal models. The present study was aimed at investigating the effect of selenium on suppression of apoptosis during vitrification-thawing process of mice ovary via studying expression of apoptosis-related genes, and also, we aimed to design statistical models for the roles of single genes and gene-gene interactions in suppression of apoptosis. METHODS: A total of 10 right ovary samples from 10 mice were randomly divided into two groups of selenium treatment (at dose 5 µg/ml sodium selenite, through adding to the media) and control group. Vitrification-thawing process was done according to the existed protocols. Real-time PCR was used for gene expression study. The apoptosis gene profile included P53, Bax, Fas, and Bcl-2. General linear model was applied to study single gene associations and gene-gene interactions. RESULTS: From the studied genes, P53 showed a significant downregulation in the selenium group in comparison to the control group (∆∆CT = 1.96; P = 0.013; relative expression (RE) = 0.28). Bcl-2 showed a significant upregulation in the selenium group in comparison to the control group (∆∆CT = -2.49; P < 0.001; RE = 3.49). No significant result was found for other genes. According to the multiple models, Bcl-2 showed a protective single gene association (beta = -0.33; P = 0.032), and Fas∗Bcl-2 interaction was significantly positive (beta = 0.19; P = 0.036). CONCLUSION: Addition of selenium to cryomedia of vitrification-thawing process could reduce the apoptosis induced by freezing-thawing stress in mice ovary via downregulation of P53 and upregulation of Bcl-2 at transcription level. Multivariable statistical models should be performed in future researches to study biological systems.

A Stereological Study of the Toxic Effects of Cerium Oxide during Pregnancy on Kidney Tissues in Neonatal NMRI Mice.

BACKGROUND: Both antioxidant and prooxidant activities have been previously reported for cerium oxide (CeO2). The aim of this study was to investigate the effects of CeO2 at different doses on changes in kidney tissues and markers in neonatal mice. METHODS: We randomly divided 30 pregnant NMRI mice into five groups (n = 6 per group)-a control group and four groups treated with intraperitoneal (i.p.) administration of different doses of CeO2 (10, 25, 80, or 250 mg/kg body weight (bw)) on gestation days (GD) 7 and GD14. At the end of the treatment period, we analyzed the kidney tissues and serum samples. The levels of two serum redox markers, malondialdehyde (MDA) and ferric reducing/antioxidant power (FRAP), were determined. Data were analyzed using one-way ANOVA and Tukey's test, and a P value of <0.05 was considered significant. RESULTS: The mean total volumes of the renal corpuscle, glomeruli, and Bowman's capsule membranes significantly increased, and there was a significant decrease in the mean total volume of Bowman's space in the high-dose CeO2 group compared to that in the control group. No statistically significant differences existed in the serum levels of MDA and FRAP in the treated and control groups. CONCLUSION: Our results suggest that high doses of CeO2 impair fetal renal development in pregnant mice, which results in kidney damage. Therefore, CeO2 administration during pregnancy could have dose-dependent adverse effects on the developing kidneys in neonates.

Effects of pomegranate peel extract on histopathology, testosterone levels and sperm of testicular torsion-detorsion induced in adult Wistar rats.

Background In the present study, effects of pomegranate peel extract have been evaluated on decreasing the damage induced by testis torsion. Methods In this study, 30 adult Wistar rats were randomly divided into three groups of control, experimental (1) and experimental (2). CONTROL: no ischemia, received vehicle alone, exposed to sham operation. Experimental (1): Received the vehicle alone during ischemia followed by 60 days' reperfusion. Experimental (2): After performing ischemia reperfusion, 500 mg/kg of pomegranate peel extract has been used for 60 days. Blood samples and sperm samples were collected. Testes were harvested and stained with haematoxylin and eosin to study the structure of seminiferous tubules. Results The statistical comparison between sperm count and their viability and testosterone hormone amount showed a significant difference between control and experimental (1) groups and control and experimental (2) groups. The results showed an improvement of morphological condition of seminiferous tubules. Conclusions Pomegranate peel extract has revealed desirable changes on the effective parameters in infertility.

Ovarian stimulation by exogenous gonadotropin decreases the implantation rate and expression of mouse blastocysts integrins.

BACKGROUND: Integrins are heterodimeric glycoprotein receptors that regulate the interaction of cells with extracellular matrix and may have a critical role in implantation. The aim of this study was to investigate the effect of ovulation induction on the expression of α4, αv, ß1, and ß3 integrins in mouse blastocyst at the time of implantation. METHODS: The ovarian stimulated and non-stimulated pregnant mice were sacrificed on the morning of 5th day of pregnancy. The blastocysts were collected, and the expression of αv, α4, ß1, and ß3 integrins was examined using real-time RT-PCR and immunocytochemical techniques, then their ovarian hormones were analyzed at the same time. The implantation sites in uterine horns of other pregnant mice in both groups were determined under a stereomicroscope on the 7th day of pregnancy. RESULTS: The results showed that the expression of αv, ß1, and ß3 integrins in both mRNA and protein levels was significantly lower in the ovarian stimulated group than the control group, and the maximum ratio of expression was belonged to ß1 molecule (P>0.05). CONCLUSION: The implantation rate in superovulated mice was significantly lower than control mice. It was suggested that ovulation induction decreased the expression of αv, ß1, and ß3 integrins of mouse blastocysts.

Ultrastructural changes of corpus luteum after ovarian stimulation at implantation period.

BACKGROUND: To achieve multiple oocytes for in vitro fertilization, ovulation induction is induced by gonadotropins; however, it has several effects on oocytes and embryo quality and endometrium receptivity. The aim of this study was to assess ultrastructural changes of corpus luteum after ovarian induction using human menopausal gonadotropin (HMG) and human chorionic gonadotropin (HCG) during luteal phase at implantation period. METHODS: Female NMRI mice (6-8 weeks) were divided into control and stimulated groups. In the control group, the mice were rendered pseudopregnant and in the ovarian induction group, the mice were rendered pseudopregnant after the ovarian induction. The samples were obtained from the ovary in each group at the same time during luteal phase at implantation period. Ultrastructural changes were assessed using electron microscopy study. RESULTS: Our results displayed some identifiable changes in ultrastructure of corpus luteum in ovarian induction group. These changes included enhancement of the apoptosis and intercellular space, whereas the angiogenesis was decreased. The findings indicated a decline in organelle density in the cytoplasm of ovarian induction, such as mitochondria, endoplasmic reticulum and polyribosome. Furthermore, chromatin condensation of nuclei was observed in some cells. CONCLUSION: The ovarian induction using HMG and HCG resulted in some ultrastructural changes on the corpus luteum at implantation period, which could affect on the pregnancy rate.

Effect of ovarian stimulation on the endometrial apoptosis at implantation period.

BACKGROUND: Apoptosis is a process that plays an important role during early stage of implantation. The aim of this study was to investigate the incidence of apoptosis in mice endometrium after ovarian stimulation at implantation period. METHODS: NMRI female mice were divided into two groups: 1) control group, which were rendered pseudopregnant by vaginal stimulation and 2) experimental group, which were stimulated using an intrapritoneal injection of 10 IU hMG followed by another injection of 10 IU hCG after 48 h. In the evening of the second injection, the mice were rendered pseudopregnant the same as control group. Samples were obtained from 1/3 middle part of uterine horns during implantation period. Apoptosis was assessed in two groups at implantation period using light and electron microscopic studies, TUNEL staining and semiquantitative RT-PCR. RESULTS: Our morphological and ultrastructural results showed apoptosis in both groups, while TUNEL analysis showed that the percentage of TUNEL-positive cells was higher in stimulated group than in the control group (P≤0.05). The expression of P53, Fas and FasL mRNA was similar in two groups but Bax and Bcl2 were much higher in control group than in the stimulated group (P≤0.05). The ratio of Bax/Bcl2 expression was much higher in stimulated group than in the control group (P≤0.05). CONCLUSION: The ovarian stimulation could change the expression of some apoptosis-related genes and enhance the incidence of endometrial apoptosis at implantation period; thus, it could affect on the implantation rate and endometrial receptivity.

The histological characteristics of cultured oral epithelium in different culture conditions.

BACKGROUND: This study was undertaken to establish the characterization of cultured oral mucosal epithelium and introducing them as an alternative source for reconstruction of ocular surface disease. METHODS: Human oral epithelial cells were cultured on simple media (DMEM/HF12) as control and co-cultured on mitomycin C-treated 3T3 feeder layer, on the amniotic membrane (AM) without nitrocellulose and the mitotically inactivated 3T3 fibroblast, and on the sandwich layer of AM fastened on the nitrocellulose as insert and 3T3 fibroblast. After 3 weeks, the characteristics of the cells were assessed morphologically and also ultrastructurally using scanning electron microscopy and transmission electron microscopy and immuno-cytochemically. RESULTS: The epithelial cells were cultured on AM spread on nitrocellulose insert and 3T3 feeder layer showed better growth than other groups and all groups of study were shown similar characteristics. The cultured oral epithelial shared the characteristics with corneal epithelium. CONCLUSION: Thus the oral epithelial could be an alternative source for transplantation.