Melatonin improves development of early mouse embryos impaired by actinomycin-D and TNF-alpha

Melatonin, a reactive oxygen species [ROS] scavenger and an antioxidant, has been shown that can inhibit apoptosis. Administration of melatonin may improve embryo development in assisted reproductive technology [ART]. The aim of this study was to evaluate the role of melatonin in inhibition of spontaneous and induced apoptosis by Tumor Necrosis Factor Alph [TNF-alpha] and actinomycin-D during preimplantation development of mouse embryos. Female BALB/c mice were superovulated with pregnant mare serum gonadotropin [PMSG] followed by human chorionic gonadotropin [HCG], then allowed to mate with male mice. The resultant 2-cell embryos were divided into six groups as follows: control [group I], melatonin [group II], actinomycin-D [group III], actinomycin-D + melatonin [group IV], TNF-alpha [group V], and TNF-alpha + melatonin [group VI]. We recorded the numbers and developmental rates of the 4-cell, 8-cell, morula and blastocyst embryos. Blastocysts were stained with acridine orange in order to assess for the embryo quality. The group IV showed a significantly higher developmental rate of blastocysts compared to group III [p<0.05]. The number of dead blastomers was significantly decreased in group IV in comparison to group III [p<0.05]. Both V and VI groups had a lower developmental rate and lesser quality of blastocysts compared with group I. There was no significant difference in the developmental rate of blastocysts from group II compared to group I [p<0.05]. Supplementation of embryo culture media with melatonin can improve the quality and developmental rate of embryos. Melatonin can prevent cell death that was induced by TNF- alpha and actinomycine-D

[Effects of knotweet or polygonum aviculare herbal extract on proliferation of HeLa cell line]

Cervical cancer is the most common cancer in women living in developing countries. Recently, for treatment of diseases such as cancer, herbal medicine is used as a supplementary. The aim of this study was assessment of anticancerous effects of polygonum aviculare herbal extract on Hela cervical cancer cell line. HeLa cells were cultured in RPMI - 1640 with 10% Fetal Bovine Serum in 5% Co2 and at 37 [degree sign] C in different concentrations [0, 0.005, 0.05, 0.01, 0.025, 0.075, 0.1, 0.125, 0.15, 0.175, 0.2, 0.25, 0.3, 0.35, 0.5, 5 mg/ml] of polygonum aviculare. For assessment of viability of cells, trypan blue staining was performed. MTT assay was used for proliferation detection. Our results showed that in 0.15, 0.20 and 0.35 mg/ml proliferation of HeLa cells decreased according to MTT assay. It was proved that polygonum aviculare had antioxidant component and could be a scavenger of free radical. Because of high production of free radicals in diseases such as cancer, the use of the herbal medicine with high amount of antioxidant could be a supplementary treatment in cancer and other diseases

High frequency electromagnetic field induces lipocalin 2 expression in mouse liver

Neutrophil gelatinase-associated lipocalin [NGAL/Lcn2], comprise a group of small extracellular proteins with a common beta-sheet-dominated 3-dimensional structure. In the past, it was assumed that the predominant role of lipocalin was acting as transport proteins. Recently it has been found that oxidative stress induces Lcn2 expression. It has been also proved that electromagnetic field [EMF] produces reactive oxygen species [ROS] in different tissues. Expression of Lcn2 following exposure to electromagnetic field has been investigated in this study. Balb/c mice [8 weeks old] were exposed to 3 mT, 50 HZ EMF for 2 months, 4 hr/day. Afterwards, the mice were sacrificed by cervical dislocation and livers were removed. The liver specimens were stained with Haematoxylin-Eosin [H and E] and analyzed under an optical microscope. Total RNA was extracted from liver and reverse transcription was performed by SuperScript III reverse transcriptase with 1 micro g of total RNA. Assessment of Lcn2 expression was performed by semiquantitative and real time-PCR. The light microscopic studies revealed that the number of lymphocyte cells was increased compared to control and dilation of sinosoids was observed in the liver. Lcn2 was up-regulated in the mice exposed to EMF both in mRNA and protein levels. To the extent of our knowledge, this is the first report dealing with up-regulation of Lcn2 in liver after exposure to EMF. The up-regulation might be a compensatory response that involves cell defense pathways and protective effects against ROS. However, further and complementary studies are required in this regards

Effects of leukemia inhibitory factor on gp130 expression and rate of metaphase II development during in vitro maturation of mouse oocyte

Leukemia inhibitory factor [LIF] is a 45-56 kDa glycoprotein that has an important role in proliferation and embryo implantation. Its effect on oocyte maturation and how to exert the function remained to be elucidated. Immature mice superovulated with human menopausal gonadotropin and germinal vesicle [GV] oocytes were obtained from ovary 48 hours after. GV oocytes were cultured in tissue culture medium 199 with 0, 100, 500 and 1,000 U/ml LIF. Cumulus expansion and in vitro maturation [IVM] rate were assessed after culture. For reverse transcriptase PCR, total RNA from GV and metaphase II [Mil] oocytes were extracted by Trizol reagent. The quantity and quality of RNA were determined by spectrophotometry and electrophoresis, respectively. Reverse transcription was performed by Super Script III reverse transcriptase with 1 micro g of total RNA followed by DNase I treatment and heat inactivation. Expression of gp130 was determined by RT-PCR. Our results showed that cumulus expansion was improved with 1,000 U/ml in culture medium compared to others. GV breakdown and Mil rate in groups with LIF were higher than control group and were dose dependant. In 1,000 U/ml, LIF rate of Mil was significantly higher than control group [P<0.05]. Our results also showed that gp130 is expressed neither in GV nor in Mil oocytes during IVM of mouse oocytes. gp130 is expressed in human oocyte but not in mouse. Our results suggest that in mouse, LIF could affect the oocyte via another receptor or via cumulus cells; however, further studies are warranted