Dietary supplementation with astaxanthin may ameliorate sperm parameters and DNA integrity in streptozotocin-induced diabetic rats / 대한생식의학회지

OBJECTIVE: Diabetes mellitus (DM) is known to cause many systemic complications as well as male infertility. Astaxanthin (ASTX) is a powerful antioxidant that is involved in a variety of biologically active processes, including those with anti-diabetes effects. The present study investigates the effect of ASTX on the spermatozoa function in streptozotocin (STZ)-induced diabetic rats. METHODS: We divided 30 adult rats into three groups (10 rats per group), with a control group that received corn oil mixed with chow. DM was induced by intra-peritoneal injection of STZ. Eight weeks after the STZ injection, half of the diabetic animals were used as diabetic controls, and the rest were treated with ASTX for 56 days. Then the parameters and chromatin integrity of the epididymal sperm were analyzed using chromomycin A3, toluidine blue (TB), and acridine orange (AO) staining. RESULTS: The count, viability, and motility of the epididymal sperm were decreased significantly in the STZ group in comparison with the control group (count and viability, p<0.001; motility, p<0.001;0.01). ASTX increased normal morphology and viable spermatozoa compared to the STZ group (morphology, p=0.001; viability, p<0.001;0.05). The percentage of abnormal chromatins in TB and AO staining was higher in the STZ group compared to the control group (p<0.001;0.001). The mean percentage of TB and AO positive spermatozoa in STZ rats was significantly lower in the STZ+ASTX group (TB, p=0.001; AO, p<0.001;0.05). CONCLUSION: This study observed that in vivo ASTX treatment partially attenuates some detrimental effect of diabetes. Conversely, ASTX improved sperm viability, normal morphology, and DNA integrity.

Effect of a high fat diet on ovary morphology, in vitro development, in vitro fertilisation rate and oocyte quality in mice

<p><b>INTRODUCTION</b>The aim of this study was to determine the effect of a high-fat diet (HFD) on oocyte maturation and quality in a mouse model.</p><p><b>METHODS</b>Female BALB/c mice were allocated to one of the following groups: (a) control group (n = 40), which received a controlled diet; or (b) HFD group (n = 40), which received an HFD for 12 weeks. Sections of the ovary were examined histologically. The number of follicles and corpora lutea were counted. In vitro maturation and in vitro fertilisation (IVF) were assessed in germinal vesicle (GV) and metaphase II (MII) oocytes, respectively. The expression of bone morphogenetic protein 15 (BMP15) and leptin receptor genes in GV and MII oocytes was evaluated using reverse transcription real-time polymerase chain reactions.</p><p><b>RESULTS</b>In the HFD group, there was a decreased number of primordial and Graafian follicles, as well as corpora lutea (p < 0.05). The rate of oocyte development to the MII stage was also reduced (p < 0.001). Cumulus expansion was observed more frequently in the control group than the HFD group (p < 0.05). The IVF rate in the HFD group was lower than that in the control group (p < 0.05). In the HFD group, BMP15 and leptin receptor genes were upregulated in the GV stage (p > 0.05) and MII stage (p < 0.05), compared to the control group.</p><p><b>CONCLUSION</b>An HFD reduces folliculogenesis in the primordial and Graafian stages, in vitro maturation and in vitro fertilisation rates, as well as oocyte quality in mice.</p>

Effect of myomectomy on endometrial glutathione peroxidase 3 [GPx3] and glycodelin mRNA expression at the time of the implantation window

In fertile women, glycodelin and glutathione peroxidase 3 [GPx3] genes expression rises during the luteal phase, with a peak occurring during the implantation window. The expression of these genes decreases in women with myomas. To determine whether myomectomy would reverse glycodelin and GPx3 expression, we evaluated the transcript levels of these genes in the endometrium of patients before and after myomectomy. Expression of glycodelin and GPx3 genes were examined prospectively during the midluteal phase in the endometrium obtained from infertile women with myoma [n = 12] before and three months after myomectomy. Endometrial expression of these genes was evaluated using quantitative real-time RT-PCR. Endometrial glycodelin mRNA expression levels [normalized to 18S rRNA expression] were increased significantly in endometrium of patients after myomectomy [P = 0.02]. GPx3 mRNA expression was increased insignificantly after myomectomy [P = 0.43]. The results showed that myomectomy increased endometrial glycodelin [significantly] and GPx3 [not significantly] gene expression after 3 months. Study at different times and detecting expression of these genes can reveal more details

Protective effects of antioxidants on sperm parameters and seminiferous tubules epithelium in high fat-fed rats

Prescription of antioxidants might increase the quality of sperm parameters and improve the rate of pregnancy in obese people who suffer from infertility. Therefore, the present study investigated protective effects of vitamin A, E and astaxanthin on sperm parameters and seminiferous tubules epithelium in high-fat diet model. Thirty-six numbers of 3 months old albino Wistar rats were divided to control, high-fat diet and high-fat diet with antioxidants groups. After 12 weeks, levels of LDL-C and HDL-C were detected in the groups. Sperm was obtained from the tail of epididymis and its parameters [count, vitality, motility and morphology] were analyzed. Testes were fixed in 10% formalin and after tissue processing, stained with Hematoxylin and Eosine [H and E] for histological evaluation. Data were analyzed by a one-way ANOVA and p<0.05 was considered significant. Our results indicated that viability, motility and normal morphology of sperm in high-fat diet [HFD] decreased significantly compared to high-fat diet with antioxidant [HFD A] and the control groups [p<0.05]. Also spermatogonium and the number of Sertoli cells increased significantly in HFD+A compared to the control [p<0.05]. As it is shown in our study, application of antioxidants decreased serum triglyceride, cholesterol and HDL-C/LDL-C in high-fat diet model and improved the semen parameters. Therefore, it is suggested that the low quality of sperm can be improved in obese men through antioxidant prescription. Finally, it seems that the antioxidants in obese patients with subfertility or infertility is a new and efficient strategy with few side effects

[Endometrial morphology following repeated ovarian stimulation in mouse]

Gonadotropins are commonly used for superovulation in human and animals to retrieve more oocyte and increase chance of pregnancy. Ovarian stimulation in assisted reproduction technology produces lower implantation rates per embryo transferred than natural and ovum donation cycles, suggesting suboptimal endometrial development due to the hormones used to recruit more oocytes. Due to the frequent use of gonadotropins in the treatment of infertility in successive periods, the aim of this study was to determine the endometrial changes in response to repeated ovarian stimulation. This experimental interventional study has done in research center of Hamadan university of medical sciences in 2012. NMIRI female mice six weeks old were used in this study and divided into 7 groups [5 each]. The mice in group 1, 2, 3 and 4 received 1, 2, 3 and 4 times pregnant mare serum gonadotropin [PMSG] and 48 hours later 7.5 IU human chorionic gonadotrophin [hCG] respectively. For each group were considered a control group which received on time gonadortropin injection. The mouses were killed 13-16 hours after hCG injection and middle part of uterine horn cut for histological study using Hematoxylin and Eosin staining. The parameters that studied were surface epithelium of endometrium, glandular epithelium, and endometrial height and axis of uterine gland. Our results showed that there are no significant differences in glandular epithelium, axis of glands and height of endometrium in experimental groups [P>0.05]. The height of surface epithelium showed significant increases after ovarian stimulation in experiment group [P0.03]. Our results showed that there are no significant differences in glandular epithelium, axis of glands, height of endometrium and height of surface epithelium between control groups and also it's experimental group [P>0.05]. Conclusion Changes in the height of surface epithelium could be one of the reasons for decrease implantation rates with repeated ovarian stimulation

Effect of intramural myomectomy on endometrial HOXA10 and HOXA11 mRNA expression at the time of implantation window

HOXA11 and HOXA10 are expressed in endometrium throughout the menstrual cycle and show a dramatic increase during the mid-luteal phase at the time of implantation. The expression of these genes is decreased in women with myomas. To determine whether myomectomy would reverse HOXA11 and HOXA10 expression, we evaluated the transcript levels of these genes in the endometria of patients before and after myomectomy. Expression of HOXA11 and HOXA10 were examined prospectively during the midluteal phase in endometrium obtained from infertile women [n=12] with myoma before and three months after myomectomy. Endometrial HOXA11 and HOXA10 expression were evaluated using quantitative real-time reverse transcriptase-polymerase chain reaction [RT-PCR]. Endometrial HOXA11 and HOXA10 mRNAs expression levels [normalized to 18SrRNA] were increased insignificantly in endometrium of patients after myomectomy [p=0.7 and p=0.15 respectively]. The results suggest that the alteration in expression pattern of these genes could not account for some aspects of fertility after myomectomy

[Semi-quantitative analysis of endometrial HOXA10 and BTEB1 mRNA expressions in the implantation window of patients with endometriosis and myoma]

The techniques used in assisted reproductive technologies have progressed considerably, but many embryos do not implant after transfer upon the use of these techniques. One of the causes of infertility is repeated implantation failure due to decreased endometrial receptivity. Furthermore, in clinical conditions such as endometriosis and myoma, implantation decreases after embryo transfer. In this case-control study the expression patterns of HOXA-10 and BTEB1 mRNAs were evaluated at the time of implantation in patients with myoma and endometriosis. In this study performed in Hamadan University of Medical Sciences during 1389, the cases included 16 patients with endometriosis and myoma [8 in each group] and the control group consisted of 8 fertile women. Endometrial sampling was performed at mid-secretory phase. Later, the expression patterns of HOXA-10 and BTEB1 mRNAs were evaluated using a semi-quantitative RT-PCR method. The optimal PCR cycles determined were 30, 32 and 26 for HOXA10, BTEB1 and beta-actin, respectively. Endometrial HOXA-10 and BTEB1 mRNA expression levels [normalized to beta-actin expression] at the time of implantation were significantly decreased in the endometrium of infertile patients with endometriosis compared with that of healthy fertile controls [P<0.05]. A similar pattern was seen in patients with myomas for both HOXA10 and BTEB1 genes, [P<0.05]. It seems that lower expression of HOXA-10 and BTEB1 mRNAs in the implantation window of endometrium that increase normally, could account for some aspects of infertility in patients with endometriosis and myoma

Semi-quantitative analysis of HOXA11, leukemia inhibitory factor and basic transcriptional element binding protein 1 mRNA expression in the mid-secretory endometrium of patients with endometriosis

HOXA11 and leukemia inhibitory factor [LIF] and basic transcriptional element binding protein1 [BTEB1] are expressed in endometrium throughout the menstrual cycle and show a dramatic increase during the mid-luteal phase at the time of implantation. In this case-control study, the expression pattern of these mRNA was evaluated in patients with endometriosis at the time of implantation. We also describe a semi-quantitative RT-PCR protocol optimized in our laboratory. Eight patients with endometriosis were considered as our case and 8 fertile women as control group. Expression levels of HOXA11, LIF and BTEB1 mRNA were measured in endometrium during the mid-secretory phase using semi-quantitative RT-PCR. We describe the detailed procedure for the analysis of HOXA11, LIF and BTEB1 mRNA levels. Endometrial HOXA11 and LIF mRNA expression levels [normalized to beta -actin expression] were significantly decreased in endometrium of infertile patients with endometriosis compared with healthy fertile controls at the time of implantation [P<0.05]. A similar trend was seen in BTEB1 mRNA expression. The results suggest that the alteration in expression pattern of the some genes could account for some aspects of infertility in endometriosis

Time course of degradation and deadenylation of maternal c-mos and cyclin A2 mRNA during early development of one-cell embryo in mouse

Early in the development of many animals, before transcription begins, any change in the pattern of protein synthesis is attributed to a change in the translational activity or stability of mRNA in the egg and early embryo. As a result, translational control is critical for a variety of developmental decisions, including oocyte maturation and initiation of preimplantation development. In this study, using real-time RT-PCR method, we defined the time course of degradation and deadenylation of an oocyte specific gene [c-mos] more precisely and a gene that is re-synthesized after ZGA [cyclin A2]. Our data indicate that oocyte-specific transcript, c-mos, degrades rapidly while cyclin A2 mRNA does not and the deadenylation of c-mos mRNA precedes the process of degradation. Our findings suggest that time-dependent elimination of different maternal mRNA is a way for regulation of translation in early development of mouse embryos