Effect of progesterone administration on tissue mast cell population and histamine content in mice uterus after ovulation induction.

OBJECTIVE: Mast cell population and histamine affect on blastocyst implantation. This study aimed to evaluate the effects of progesterone administration after induction of ovulation on the uterine tissue mast cell population and histamine content in mice. METHODS: We ran an experimental study on three groups of mice; control group, ovulation induction (induction group), and ovulation induction along with progesterone administration (progesterone group). Mast cells were counted using toluidine blue staining, and the histamine level was measured through spectrophotometry. RESULTS: According to the analysis of variance (ANOVA), there was no difference in mast cell population in endometrium (p=0.138) nor in myometrium (p=0.611). The ratio of mast cells in the myometrium per endometrium increased in the progesterone group in comparison to the control group based on a generalized linear model (p=0.041). The uterine histamine level was different between the groups, based on the ANOVA (p=0.039), in which the progesterone group had lower amounts of histamine. CONCLUSIONS: Progesterone administration after ovulation induction did not decrease the number of endometrial mast cells and could have increased the ratio of myometrium mast cells per endometrium mast cell. The histamine level in uterus decreased by the administration of progesterone in the ovulation-induced mice.

Protective Effect of Thyme Honey against Valproic Acid Hepatotoxicity in Wistar Rats.

INTRODUCTION: Valproic acid is a medication most commonly used in the treatment of emotional and neurological depression, psychological imbalances, epilepsy, and bipolar disorder. Dark honey, like thyme honey, contains more antioxidant compounds than other samples. The purpose of this study was to evaluate the effect of thyme honey on the potential hepatic effects of valproic acid. METHODS: In this study, 48 male rats were randomly divided into 8 groups (n = 6): G1 (control): healthy rats (normal saline 0.9%), G2: thyme honey (1 g/kg), G3: thyme honey (2 g/kg dose), G4: thyme honey (3 g/kg dose), G5: VPA (500 mg/kg), G6: VPA (500 mg/kg) and thyme honey (1 g/kg), G7: VPA (500 mg/kg) and thyme honey (2 g/kg dose), and G8: VPA (500 mg/kg) and thyme honey (3 g/kg dose). Groups G1 to G5 received the drug for 28 days. On day 14, administration of thyme honey for G6 to G8 groups was carried out using gavage until day 28. VPA was administered one hour after honey. To carry out the biochemical evaluation, blood samples were collected from all the groups and their serums were used for MDA, TAC, and liver enzymes (AST, ALT, and GGT). Tissue samples of each rat were also removed for histological studies with hematoxylin-eosin and Masson's trichrome staining. RESULTS: The use of thyme honey significantly improved the histopathological parameters of the liver tissue, including hypertrophic degeneration and nucleus alteration, expansion of sinusoids, fibrosis and hepatic necrosis, and inflammation as well as hypertrophy of Kupffer cells. In the groups receiving VPA, the rate of lipid peroxidation increased, which indicates the destruction of the liver cell membrane due to drug consumption. TAC levels also increased following increase in thyme honey dosage (p ≤ 0.05). The results of liver enzyme analysis showed a decrease in AST and ALT levels in the G6 group and a decrease in GGT level in the G8 group (p ≤ 0.05). CONCLUSION: Based on the results of this study, it seems that high percentage of antioxidants in thyme honey enabled it to improve hepatic complications and reduce the rate of hepatocellular destruction.

Pre-Implantation Effects of Progesterone Administration on Ovarian Angiogenesis after Ovarian Stimulation: A Histological, Hormonal, and Molecular Analysis.

OBJECTIVE: Progesterone (P4) is known to directly affect ovarian tissue angiogenesis. The present study was designed to show how P4 affects ovarian angiogenesis in hormonal, histological, and molecular levels. METHODS: Fifteen adult female NMRI mice were divided into three groups: Control Group; Case Group I (ovarian stimulation alone); and Case Group II (ovarian stimulation followed by P4 administration). Blood and ovarian tissue samples were assessed for hormonal, histological, and molecular alterations. Gene expression for ovarian vascular endothelium growth factor (VEGF) and hypoxia-inducible factor-1 alpha (HIF-1α) was analyzed using real-time PCR. RESULTS: Ovarian hormone levels were increased in the case groups compared with the control group (p<0.05). Quantitative corpus luteum parameters were increased in the case groups compared with the control group (p<0.05). Quantitative ovarian vascular parameters were significantly different in the case groups compared with the control group. Gene expression analyses shows that the mice in Case Group I had higher levels of ovarian VEGF expression than the mice in the control group (p<0.05). No significant difference in gene expression was observed for HIF-1ɑ. CONCLUSION: Treatment with P4 after ovarian stimulation enhanced ovarian angiogenesis by increasing hormone levels and causing significant structural changes.

Effect of selenium on freezing-thawing damage of mice spermatogonial stem cell: a model to preserve fertility in childhood cancers.

BACKGROUND: During treatment of childhood cancers, fertility of boys may be affected. Therefore, freezing spermatogonial stem cell (SSC) is recommended. However, freezing-thawing process may cause damage to SSCs. This study was conducted to evaluate protective effects of selenium on freezing-thawing damage of mice SSCs using investigation of cell viability and investigation of apoptosis related genes expression including Fas, Caspase3, Bcl2, Bax and P53. METHODS: SSCs were extracted from 80 6-day-old mice. The SSCs were divided into four groups: cryopreservation along with selenium (low and high dose), vitrification along with selenium (low and high dose), cryopreservation control, and vitrification control. Trypan blue staining and real-time polymerase chain reaction (real-time PCR) were used to investigate cell viability and gene expression, respectively. RESULT: Comparison of cell viability in the experimental groups did not show a significant association. Expression of Fas and Caspase3 was significantly lower in cryopreservation group with low-dose selenium. Expression of Bcl2 was significantly lower in cryopreservation group with high-dose selenium. Expression of Bax and Caspase3 was significantly lower in vitrification group with low-dose selenium, and expression of P53 was significantly upper. Expression of Bax and Fas was significantly lower in vitrification group with high-dose selenium, and expression of P53 was significantly upper (P<0.001). CONCLUSIONS: Selenium had dose dependent effect on apoptosis related genes profile. The only evident effect was the effect of low-dose selenium in cryopreservation on inhibition of apoptosis via extrinsic pathway.

Coenzyme Q10 protects skeletal muscle from ischemia-reperfusion through the NF-kappa B pathway.

OBJECTIVE: Coenzyme Q10 (CoQ10) has antioxidant and anti-inflammatory activity. The aim of this study was to investigate the effects of CoQ10 on the inhibition of nuclear factor-kappa B (NF-κB) activation during ischemia-reperfusion (I/R) of skeletal muscle. METHODS: For ischemia induction, the animals were anesthetized and the external iliac vessels blocked for three hours. CoQ10 or vehicle was given intraperitoneally during ischemia, just before reperfusion. Four groups received 3,7,14 and 28 days' reperfusion, respectively, after the intraperitoneal injection of CoQ10 and four corresponding groups received vehicle only. After reperfusion, the gastrocnemius muscles were removed, fixed and stained for the analysis of edema and mast cell infiltration. RESULTS: Immuno-histochemistry staining was performed for the detection of tumor necrosis factor alpha (TNF-α) and NF-κB. CoQ10-treated groups showed a significant decrease of mast cell infiltration in the gastrocnemius muscle and edema as compared with the corresponding non-treated groups. Also, CoQ10-treated groups showed a significant TNF-α and NF-κB expression decrease when compared to the corresponding non-treated controls. The results of this study showed CoQ10 administration with ischemia decreased interstitial edema, degeneration of muscle fibers and infiltration of mast cells. CONCLUSIONS: It seems that CoQ10 has inhibitory effects on NF-κB and TNF-α activation.

The effect of aloe vera on the expression of wound healing factors (TGFß1 and bFGF) in mouse embryonic fibroblast cell: In vitro study.

BACKGROUND: Aloe vera (A.v) have been used traditionally for topical treatment of wounds and burns in different countries for centuries, but the mechanism of this effect is not well understood. Various growth factors are implicated in the process of wound healing. Among the different growth factors involved in the process, TGFß1 and bFGF are the most importantly expressed in fibroblast cells. The aim of this study was to evaluate the effect of A.v on the expression of angiogenesis growth factors in mouse embryonic fibroblast cells. METHODS: We exposed mouse embryonic fibroblast cells to different concentrations of A.v (50, 100 and 150µg/ml) at two different time of 12 and 24h. Fibroblast cell without A.v treatment serves as the control. The expression of TGFß1and bFGF was measured by real time-polymerase chain reaction (real-time-PCR) and enzyme-linked immunosorbent assay (ELISA) at the level of gene and protein. RESULTS: We observed that A.v gel at first up-regulated the expression of TGFß1 and bFGF, but, these genes were later repressed after a particular time. CONCLUSION: Our results demonstrated that A.v was dose-dependent and time-dependent on the expression of bFGF and TGFß1 in fibroblast cell in vitro. This mechanism can be employed in the prospective treatment of physical lesion.

Quercetin ameliorates peripheral nerve ischemia-reperfusion injury through the NF-kappa B pathway.

This study aimed to investigate the protective effect of quercetin against ischemia-reperfusion (IR) injury induced in the sciatic nerve of the rat. Quercetin (20 mg/kg) was given during ischemia just before reperfusion. Four groups of rats (Q+IR3, Q+IR7, Q+IR14, and Q+IR28) received 3, 7, 14, and 28 days of reperfusion, respectively, after the intraperitoneal injection of quercetin. After reperfusion, a behavioral test was performed and the sciatic functional index was calculated. Each sciatic nerve was stained to check for edema and ischemic fiber degeneration. Immunohistochemical staining was performed to detect TNF-alpha and NF-kappa B, and TUNEL staining was carried out to detect apoptosis. The Q+IR3, Q+IR7, and Q+IR14 groups showed significantly increased behavioral scores and ameliorated sciatic functional index values compared to IR-injured rats that received vehicle alone during ischemia and then the same period of reperfusion. The Q+IR3, Q+IR7, Q+IR14, and Q+IR28 groups presented significant ischemic fiber degeneration (IFD), TNF-alpha expression, and apoptosis as compared with the IR-injured and perfused rats that did not receive quercetin. The Q+IR3, Q+IR7, and Q+IR28 groups also exhibited significantly decreased NF-kappa B expression (p < 0.001, p = 0.001, p = 0.026) as compared with the IR-injured rats that were perfused but did not receive quercetin. These results imply that quercetin may be beneficial in the treatment of sciatic IR injury because of its antiapoptotic and antiinflammatory effects and its ability to decrease the expression of NF-kappa B.

The effect of progesterone treatment after ovarian induction on endometrial VEGF gene expression and its receptors in mice at pre-implantation time.

OBJECTIVES: Progestrone is a prequisite for pre-implantation angiogenesis and induce decidual angiogenesis. It is unknown the effect of progestrone administration on the endometrium of hyperstimulated mice at pre-implantation time. MATERIAL AND METHODS: Adult female NMRI mice were divided in three groups [control group, ovarian stimulated group and progestrone treated mice after ovarian stimulation]. Uterine horn samples removed at pre-implantation time in each group. Motic image Plus 2 software was used to assess the quantitative vascular parameters of endometrium. Gene expression was determined for vascular endothelial growth factor (VEGF), FMS-like tyrosine kinase (FLT) and Kinase insert domain protein receptor (FLK) genes using the real time PCR method. Data analysis was done with LinReg PCR and Rest-RG software. RESULTS: Comparison between progestrone treated mice after ovarian stimulation with control group showed that increase in rate of VEGF gene expression [0.775] and decrease in rate of FLK [6.072] and FLT [1.711] gene expression. Analysis of the data on quantitative vascular parameters were indicated remarkable increase in quantitative vascular parameters of progestrone treated mice compare to control group. CONCLUSION: Biological effect of progestrone on the vascular changes after ovarian stimulation resulted in an increase in VEGF receptors experession, it seems that induced angiogenesis by progesterone could result in better condition for implantation.

Application of gel-based proteomic technique in female reproductive investigations.

Recently, gel-based proteomics has been increasingly applied to investigate proteins involved in female reproductive tract in healthy and disease states. Gel-based proteomics coupled by mass spectrometry (MS) facilitate the identification of new proteins playing roles in cellular and molecular interactions underlying female reproductive biology and it is a useful method to identify novel biomarkers of diseases by studying thousands of proteins simultaneously involved in female reproductive tract in healthy state compared to disease state. This review will discuss the best studies areas contributed to female reproductive biology in which gel-based proteomics coupled by MS has been applied to generate proteome of female reproductive tract in a healthy state.

Effects of epigallocatechin gallate on tissue protection and functional recovery after contusive spinal cord injury in rats.

Recent studies revealed the neuroprotective effects of epigallocatechin gallate (EGCG) on a variety of neural injury .The purpose of this study was to determine the effects of EGCG on the tissue protection and behavioral improvement after spinal cord injury (SCI). Rats were randomly divided into four groups of 18 rats each as follows: sham-operated group, trauma group, and EGCG treatment groups (50 mg/kg, i.p., immediately and 1 hour after SCI). Spinal cord samples were taken 24 hours after injury and studied for determination of malodialdehyde (MDA) levels, immunohistochemistry of Bax and Bcl-2, and TUNEL reaction. Behavioral testing was performed weekly up to 6 weeks post-injury. Then, the rats were euthanized for histopathological assessment. The results showed that MDA levels were significantly decreased in EGCG treatment groups. Greater Bcl-2 and attenuated Bax expression could be detected in the EGCG-treated rats. EGCG significantly reduced TUNEL-positive rate. Also, EGCG significantly reduced the percentage of lesion area and improved behavioral function than the trauma group. On the basis of these findings, we propose that EGCG may be effective in protecting rat spinal cord from secondary injury.

Transplantation and homing of mouse embryonic stem cells treated with erythropoietin in spleen and liver of irradiated mice.

BACKGROUND: The present study was designed to evaluate the homing potential of mouse embryonic stem cells (ESC) treated with erythropoietin (EPO) in hematopoietic organs such as spleen and liver after transplantation using morphological and immuno-histochemical techniques. METHODS: Day-four embryoid body (EB)-derived cells were dissociated and re-plated in medium in the presence and absence of EPO for three days. The EPO- and untreated differentiated cells were labeled with 5-bromo-2 deoxyuridine (BrdU) before transplantation and analyzed using flow cytometry and reverse transcription-PCR methods. BrdU-labeled cells were injected via the tail vein into irradiated adult mice in both groups. The spleen colony-forming unit assay (CFU-S) was performed 12 days after transplantation. Immuno-histochemistry was also carried out to trace transplanted cells. RESULTS: The percentage of CD34 positive cells was 5.51 +/- 1.06% in the EPO-treated group and 1.63 +/- 0.225% in untreated group. The RT-PCR analysis showed that the EPO-treated cells expressed epsilon globin, betaH1 globin, RUNX1 and EPO receptor genes, but the beta-major globin gene was not expressed. The number of colonies formed in the spleens of treated group (17.33 +/- 4.726) was significantly different from the control group (6 +/- 1). The population of BrdU positive cells in spleen of EPO-treated cell-transplanted group was higher than that of the control group. Also, BrdU positive cells were observed in the central vein of the liver sections of EPO-treated and control groups but were not observed in the liver parenchyma. There were not BrdU positive cells in the spleen and liver sections of the sham group. CONCLUSION: Our results confirm that ESC have the ability to home and form colonies in spleen after transplantation and EPO-treated EB-derived cells caused an increase in the number of colonies in spleen after CFU-S.

The effect of bone morphogenetic protein 4 on the differentiation of mouse embryonic stem cell to erythroid lineage in serum free and serum supplemented media.

This study was done to compare the effects of bone morphogenetic protein-4 (BMP-4) on mouse embryonic stem cells (ESC) differentiation to erythroid lineage in serum free and serum supplemented media. The embryoid bodies (EBs) cells were seeded in semisolid serum free and serum supplemented media in the presence of different concentrations of BMP-4. The erythroid colonies were assessed morphologically, ultrastructurally and by benzidine staining. The expression of the epsilon (ε), ßH1 and ßmajor globins, Runx1 and ß2m genes was evaluated by Real time PCR. The colony size and the percent of benzidine-positive colonies increased in both BMP-4 supplementd groups but the number of colonies were lower in these groups than control. Erythropoiesis related genes were expressed in both serum free and serum supplemented groups. There were not significant differences between the ratios of genes expression to ß2m in these groups except the ratio of Runx1 was significantly higher in serum free group (P<0.05). The ratio of ε and ßH1 to ß2m in EBs was higher than both BMP-4 containing groups (P<0.05) and ßmajor was not expressed in EB cells. These findings showed in serum free condition the effects of BMP-4 on the erythroid differentiation was prominent than serum supplemented group.