ABSTRACT
Objective: Infertility is a worldwide health problem which affects approximately 15% of sexually active couples. One of the factors influencing the fertility is melatonin. Also, protection of oocytes and embryos from oxidative stress inducing chemicals in the culture medium is important. The aim of the present study was to investigate if melatonin could regulate hyaluronan synthase-2 [HAS2] and Progesterone receptor [PGR] expressions in the cumulus cells of mice oocytes and provide an in vitro fertilization [IVF] approach
Materials and Methods: In this experimental study, for this purpose, 30 adult female mice and 15 adult male mice were used. The female mice were superovulated using 10 U of pregnant mare serum gonadotropin [PMSG] and 24 hours later, 10 U of human chorionic gonadotropin [hCG] were injected. Next, cumulus oocyte complexes [COCs] were collected from the oviducts of the female mice by using a matrix-flushing method. The cumulus cells were cultured with melatonin 10 microM for 6 hours and for real-time reverse transcription-polymerase chain reaction [RT-PCR] was used for evaluation of HAS2 and PGR expression levels. The fertilization rate was evaluated through IVF. All the data were analyzed using a t test
Results: The results of this study showed that HAS2 and PGR expressions in the cumulus cells of the mice receiving melatonin increased in comparison to the control groups. Also, IVF results revealed an enhancement in fertilization rate in the experimental groups compared to the control groups
Conclusion: To improve the oocyte quality and provide new approaches for infertility treatment, administration of melatonin as an antioxidant, showed promising results. Thus, it is concluded that fertility outcomes can be improved by melatonin it enhancesPGR
ABSTRACT
Background: Linoleic acid [LA] is a polyunsaturated fatty acid present in high concentrations in follicular fluid, when added to maturation culture media, it affects oocyte competence
Objective: In the present study, we investigated effect of linoleic acid supplementation on in vitro maturation, embryo development and apoptotic related gene expression in ovine
Materials and Methods: The experiments conducted on 450 ovine Cumulus-oocyte complexes [COCs] with homogenous ooplasm and more than two compact layers of cumulus cells. For in vitro maturation COCs were randomly allocated into four treatment groups for 24 hr period. Treatment groups were as follow: control maturation media, 0 micro M LA, 50 micro M LA, 100 micro M LA and 200 micro M LA. The cumulus cell expansion and blastocysts rates were recorded. Total RNA was isolated from embryo pools, reverse transcribed into cDNA, and subjected to apoptotic gene expression by real-time PCR
Results: Highest concentration [200 micro M/mL] of LA significantly decreased the rate of fully expanded cumulus cells 24 hr after in vitro maturation [IVM] and the percentage of blastocyste rate compared with the control [p<0.05]. These inhibitory effects were associated with an increased in relative mRNA expression of Bax [Bcl-2- associated X] gene compared with controls
Conclusion: Data obtained in present study suggest that low concentration of LA used for maturation had no deleterious effect on subsequent embryonic development compared to high concentration of LA. Relative expression of Bcl-2 [B-cell lymphoma 2] and Bax in embryos seems to be associated with LA concentration
ABSTRACT
Background: Stearic acid is known as a potent anti-inflammatory lipid. This fatty acid has profound and diverse effects on liver metabolism. The aim of this study was to investigate the effect of stearic acid on markers of hepatocyte transplantation in rats with acetaminophen [APAP]-induced liver damage
Methods: Wistar rats were randomly assigned to 10-day treatment. Stearic acid was administered to the rats with APAP-induced liver damage. The isolated liver cells were infused intraperitoneally into rats. Blood samples were obtained to evaluate the changes in the serum liver enzymes, including activities of aspartate aminotransferase [AST], alanine aminotransferase [ALT] and alkaline phosphatase [ALP] and the level of serum albumin. To assess the engraftment of infused hepatocytes, rats were euthanized, and the liver DNA was used for PCR using sex-determining region Y [SRY] primers
Results: The levels of AST, ALT and ALP in the serum of rats with APAP-induced liver INJURY were significantly increased and returned to the levels in control group by day six. The APAP-induced decrease in albumin was significantly improved in rats through cell therapy, when compared with that in the APAP-alone treated rats. SRY PCR analysis showed the presence of the transplanted cells in the liver of transplanted rats
Conclusion: Stearic acid-rich diet in combination with cell therapy accelerates the recovering of hepatic dysfunction in a rat model of liver injury
ABSTRACT
Background: Ammonium is produced in culture medium due to amino acids degradationand has adverse effect on in vitro culture of embryo. In the current study, thepurpose was to evaluate the effects of ammuniom chloride [AC] on in vitro oocytematuration rate and early embryo development in the sheep and its effect on the expressionof Bcl-2
Methods: In vitro maturation [IVM] was performed in the presence of various concentrations[0, 29, 88,132,176 M/ml] of ammonium chloride [NH4CL][AC]. Meioticmaturation, embryonic development and expression of Bcl2 gene in Blastocystcells were determined. The data were analyzed by one-way ANOVA and Tukey postHOC test, and values with p<0.05 were considered statistically significant
Results: The highest concentration [176micro M] of AC significantly decreased the rateof fully expanded cumulus cells 24 hr after IVM compared with the control group[p<0.05]. Moreover, significantly lower rates of MII oocytes were found in the 176micro M AC group compared with the 29micro M AC group. The percentage of zygotes developingto blastocysts in 176micro M AC was lower than the other group. Also, supplementationof the oocyte maturation media with 176micro M AC decreased Bcl2 expression
Conclusion: Our results suggested that significant increase in IVM rate could be obtainedwith supplementation maturation medium with AC in a dose dependent manner.Increased AC concentration led to lower blastocyst rate under normal condition.However, regulation of pro-apoptotic [Bcl-2] gene did not change with differentconcentrations of AC supplementing
ABSTRACT
Buthionine sulfoximine is an agent that reduces intracellular glutathione and anti-oxidant enzymes and by this means is involved in pathology of some disease. Since low glutathione level may affect embryonic development, the aim of the present study is to investigate glutathione reduction in pregnant and non-pregnant mice. In the present study, the mice were divided into 4 groups [10 in each group]. The groups included pregnant and non-pregnant, each one consisting of a control and an experimental subgroup. The experimental groups received 2 mMol/kg buthionine sulfoximine [in the pregnant group on the 10th day of pregnancy], then 12 hours after buthionine sulfoximine injection, the mice in all control and experimental groups were killed and the blood obtained from their heart, and the glutathione level were determined and compared with each other. Glutathione level in experimental pregnant group was reduced significantly [p<0.05] in comparison with control pregnant group. In non-pregnant group, also the level of glutathione were reduced significantly [p<0.05] in comparison to control group. The results indicated that buthionine sulfoximine injection could reduce glutathione level both in the pregnant and non-pregnant mice. Also the effect of buthionine sulfoximine in the pregnant group was more extensive than that of non-pregnant group
ABSTRACT
Endometrial development has an important role in blastocyst adhesion and implantation. During IVF cycles, endometrial development is enhanced by progesterone. The aim of this study was to compare ultrastructural and morphometrical characteristics of mice uterine endometrium in natural cycle with those in superovulated cycles received progesterone or Sildenafil. In This study, 60 female bulb/c mice were divided into 4 groups: a control and 3 experimental; gonadotropin, gonadotropin+ Sildenafil and gonadotropin+ progesterone. In experimental groups the mice superovulated mated. In the gonadotropin+ progesterone and gonadotropin+ Viagra groups, the mice respectively received 1mg progesterone and 3 mg Sildenafil citrate. Their uterine specimens were prepared for morphometrical and ultrastructural study. Height of the epithelial cells was measured, using motic software. Statistical analysis was performed using ANOVA. Microscopy revealed that in control group the cells had numerous apical microvilli and the height of the cells was 20.52 +/- 2.43 microm. In gonadotropin+ progesterone group, the granules were found in basal and apical portions and cellular height were 17.91 +/- 2.78 microm which were significantly shorter than in the control and gonadotropin groups [p<0.001]. In this group, the apical membrane also contained pinopodes. In gonadotropin +Sildenafil group, the granules were found in both apical and basal portions and the height of the cells were 17.60 +/- 2.49 microm which were significantly shorter than in the control and gonadotropin groups [p<0.001]. In this group, pinopodes appeared slightly extensive than the other groups. It is concluded that superovulatory drugs in mice stimulate endometrial maturation but injection of Sildenafil is nearly more positive
Subject(s)
Female , Animals, Laboratory , Superovulation , Progesterone , Piperazines , Sulfones , Purines , Mice, Inbred BALB CABSTRACT
More than 40% of infertilities are due to endometriosis. Ultrustructural and histochemical study of endometrium will help to clarify the etiology of endometriosis. The aim of the present study was to investigate the ultrastructure and occurrence of apoptosis in endometrial cells of women with or without endometriosis. In the present case-control study, endometrial specimens from 12 women without endometriosis [as control] and 12 women with endometriosis [as case] were examined. Specimens for control group were obtained from the patients that were referred to gynecology hospital for hysterectomy due to various reasons. In case group the endometriosis was diagnosed according to laparoscopy and endometrial samples were taken using pippel biopsy. The specimens from both case and control groups were processed for Transmission Electron Microscopy [TEM], TUNEL reaction technique and morphometric studies. The results show that endometrial epithelium lost its continuity in women with endometriosis and endometrial cells have euchromatic nucleus in comparison to those from non-endometriosis. There were several apoptotic cells in the luminal and glandular endometrial epithelium and stroma from endometrium of control group. However, apoptotic cells were rarely seen in the endometrium from women with endometriosis. The difference in number of apoptotic cells between two groups statically was significant [p<0.001]. Regarding the ultrastructural characteristics of endometrial epithelial cells and comparison of apoptotic occurrence in control and case groups it is concluded that endometrial cells in endometriosis group have higher potential to survive and possibly implant.
Subject(s)
Humans , Female , Adolescent , Young Adult , Adult , Middle Aged , Apoptosis , Case-Control StudiesABSTRACT
Cyclo-oxygenase-2 [COX-2] specific inhibitors were examined for predication or treatment of different tumors and it is indicated that COX-2 specific inhibitors play an important regulatory role in apoptosis of tumoral tissues. Therefore, the present study was designed in order to examine the preventive effects of a COX-2 specific inhibitor called. celecoxib on 4-nitroquinoline 1-oxide [4NQO]-induced squamous cell carcinoma on rat. In this experimental study, 30 Sprague Dawley rats [with the age of 3- 3.5 months] were selected and divided into three groups. In order to induce lingual carcinoma, 4NQO powder was prepared 3 times a week for each cage. In this study, celecoxib power was mixed with a basic food [basal diet] in order to examine the systematic effect. Tongue samples were sent to laboratory for immunohistochemical [IHC] staining and histological examination. Based on morphological criteria and the ratio of apoptosis to cell proliferation, the prevalence of tongue precancerous lesions was reduced significantly by celecoxib. Celecoxib systematic has inhibitory effects on the 4-nitroquinoline 1-oxide [4NQO]-induced squamous cell carcinoma of tongue. The effect of celecoxib is probably via suppression of cell proliferation and induction of apoptosis
Subject(s)
Animals, Laboratory , Pyrazoles , Sulfonamides , Chemoprevention , Tongue Neoplasms/prevention & control , Carcinoma, Squamous Cell , Rats, Sprague-Dawley , 4-Nitroquinoline-1-oxide , Cell Proliferation/drug effects , Apoptosis/drug effectsABSTRACT
Ischemia- reperfusion is the common cause of apoptosis in most of cells specially myocytes. Prevention and reduction of apoptosis in myocardium can be one of the main medical goal before surgical operation, angioplasty and after infarction. Erythropoietin receiving effect 24 hours before hypoxia beginning on myocytes apoptosis rate and inflammatory process following half an hour hypoxia and 1.5 hour reperfusion are aim of this study. 40 Rats were divided randomly into two groups. 24 hours before surgical operation, 5000 Iu/Kg erythropoietin was injected to experimental group. During operation 12 rats from experimental group and 11 rats from control group were lost. After anesthesia, using ligation in left coronary artery for 30 minutes hypoxia and 1.5 hours reperfusion were applied. Then Thorax was opened and after bleeding, the animal's heart was isolated and two tissue samples of infarct and non-infarct area were separated and fixed. Then blood serum samples separated and incubated in -76[degree] C. Apoptosis intensity in heart tissue was measured by tunel method CK-MB level by method and DGKC, hsCRP by Elisa using Immunodiagnostic kit. The results were calculated Mean +/- SD. Then using paired student's t- test their deference were shown. Level of statistical significant was considered P< 0.05. Activity level of CK-MB [1550U/L to 340] in experimental group was less than control group [P
ABSTRACT
The purpose of this study was to compare the histocompatibility of white [WMTA] and gray [GMTA] mineral trioxide aggregate mixed with 0.12% chlorhexidine [CHX] and distilled water [DW] in subcutaneous connective tissues of rats. The freshly mixed WMTA and GMTA with CHX or DW were inserted in polyethylene tubes and implanted into dorsal subcutaneous connective tissue of 50 Wistar Albino rats; tissue biopsies were collected and were then examined histologically 7, 15, 30, 60 and 90 days after the implantation procedure. The histology results were scored from 1-4; score 4 was considered as the worst finding. Data were analyzed using one-way ANOVA tests. All experimented materials were tolerated well by the connective tissues after 90- day evaluation, except for the WMTA/CHX group that had significantly more mean inflammatory scores [P<0.001]. There was a statistically significant difference in the mean inflammation grades between experimental groups in each interval [P<0.001]. After 90 days, GMTA/CHX group had the lowest inflammatory score. Although adding CHX to WMTA produces significantly higher inflammatory response, it seems a suitable substitute for DW in combination with GMTA. Further research is necessary to recommend this mixture for clinical use