Vitrification affects the expression of matrix metalloproteinases and their tissue inhibitors of mouse ovarian tissue

Background: One of the most major obstacles of ovarian tissue vitrification is suboptimal developmental competence of follicles. Matrix metalloproteinases 2 [MMP-2] and 9 [MMP-9] and their tissue inhibitors TIMP-1 and TIMP-2 are involved in the remodeling of the extracellular matrix in the ovaries

Objective: This study aimed to evaluate the expression of MMP-2, MMP-9, TIMP-1, and TIMP-2 genes in the preantral follicles derived from vitrified mouse ovaries

Materials and Methods: In this experimental study, the gene expression of MMP-2, MMP-9, TIMP-1, and TIMP-2 in the isolated preantral follicles derived from fresh and vitrified ovaries of 14-16 days old female mice through real time qRT-PCR was evaluated. Developmental parameters, including survival rate, growth, antrum formation and metaphase II oocytes were also analyzed

Results: The developmental parameters of fresh preantral follicles were significantly higher than vitrified preantral follicles. The TIMP-1 and MMP-9 expression levels showed no differences between fresh and vitrified preantral follicles [p=0.22, p=0.11 respectively]. By contrast, TIMP-2 expression significantly decreased [p=0.00] and MMP-2 expression increased significantly [p=0.00] in vitrified preantral follicles compared with to fresh ones

Conclusion: Changes in expression of MMP-2 and TIMP-2 after ovarian tissues vitrification is partially correlated with decrease in follicle development

effect of steroid hormones on the mRNA expression of oct4 and sox2 in uterine tissue of the ovariectomized mice model of menopause

Background: The uterus is a dynamic tissue responding to hormonal changes during reproductive cycles. As such, uterine stem cells have been studied in recent years. Transcription factors oct4 and sox2 are critical for effective maintenance of pluripotent cell identity

Objective: The present research evaluated the mRNA expression of oct4 and sox2 in the uterine tissues of ovariectomized mice treated with steroid hormones

Materials and Methods: In this experimental study, adult virgin female mice were ovariectomized and treated with estradiol 17 [E2], progesterone [P4], and a combination of E2 and P4 [E2 and P4] for 5 days. Uterine tissues were removed, and immunofluorescent [IF] staining and quantitative real-time PCR of oct4 and sox2 markers were performed

Results: IF showed oct4 and sox2 expression in the uterine endometrium and myometrium among all groups. The mRNA expression of oct4 [p=0.022] and sox2 [p=0.042] in the E2-treated group significantly were decreased compared to that in the control group. By contrast, the mRNA expression of oct4 and sox2 in the P4 [p=0.641 and 0.489 respectively] and E2 and P4-treated groups [p=0.267 and 0.264 respectively] did not show significant differences compared to the control group

Conclusion: The results indicate ovarian steroid hormones change the expression of oct4 and sox2 in the mice uterine tissues, which suggest the involvement of steroid hormonal regulation in uterine stem cells

Comparing the effect of silybin and silybin advanced[TM] on viability and HER2 expression on the human breast cancer SKBR3 cell line by no serum starvation

The polyphenol silybin has anti-oxidant and anti-cancer properties. The poor bioavailability of some polyphenols [flavonoids, and terpenoids] can be improved by binding them to phosphatidylcholine [phytosome technology]. Many studies have focused on the most common phytosome, silybin-phosphatidylcholine, particularly for its hepatoprotective effects. However, in recent years, studies have also been conducted to determine its anti-cancer effect. Considering that the serum starvation should not be used for studies that are not focused on cell cycle arrest, we studied the effect of silybin-phosphatidylcholine from Silybin Advanced[TM] in 1:2 ratio [one part silybin bound to two parts phosphatidylcholine] on HER2 gene expression on the SKBR3 breast cancer cell line which were cultured in complete medium [not serum deprivation]. The results were compared with our previous study of silybin on HER2 expression on SKBR3 cells. An MTT test was used to determine concentrations for cell treatment, and the gene expression was defined by real-time RT-PCR. Outcomes showed significant concentration- and time-dependent cell growth inhibitory effects of silybin, and silybin-phosphatidylcholine and HER2 down regulation on SKBR3 cells. Silybin-phosphatidylcholine concentrations had a much larger inhibitory and HER2 down regulate effect on cell growth than the same silybin concentrations on SKBR3 cells

Cyclic AMP pathway modifies memory through neural cell adhesion molecule alterations in the rat hippocampus

Neural Cell Adhesion Molecules [NCAMs] are known to influence memory by affecting neural cell-cell and cell-extracellular matrix junctions. This study investigated the possible role of cAMP pathway in the expression of hippocampal NCAM and its polysialylated derivative [PSA-NCAM]. The following pharmacological tools were employed for manipulation of cAMP pathway: a] forskolin; the activator of adenylyl cyclase [AC], b] 8-Br-cAMP; a protein kinase A [PKA] agonist, c] 8-pCPT-2'-O-Me-cAMP; a selective enhancer of exchange protein activated by cAMP [Epac] and d] Rp-cAMP; a PKA inhibitor. Memory acquisition was tested by passive avoidance paradigm after injecting the above compounds for three consecutive days into the CA1 region of dorsal hippocampus of rats. Forskolin and 8-Br-cAMP enhanced memory retrieval while Rp-cAMP significantly reduced memory and NCAM levels. 8-pCPT-2'-O-Me-cAMP failed to alter memory performance or NCAM levels as compared to vehicle. We observed no significant changes in PSA-NCAM, however the expression of St8sia4 and St8sia2 [the polysialyltransferase isoforms] were altered. The mRNA levels of St8sia4 was down-regulated by 8-Br-cAMP, Rp-cAMP and 8-pCPT while forskolin led to almost 3 and 5 fold increase in mRNAs of St8sia2 and St8sia4, respectively. The current insight might endorse the predominant role of PKA as compared to Epac in cAMP pathway in expression of NCAM and memory function

comparison of the effects of silybin and silybin-phosphatidylcholine on viability and ESR expression in human breast cancer T47D cell line

Silybin is a polyphenol with anti-oxidant and anti-cancer properties. The poor bioavailability of some polyphenols can be improved by binding to phosphatidylcholine. In recent years, studies have been conducted to evaluate the anti-cancer effect of silybin. We studied the effect of silybin and silybin-phosphatidylcholine on ESR1 and ESR2 gene expression and viability in the T47D breast cancer cell line. In this experimental study, a 3-[4,5-Dimethylthiazol-2-Yl]-2,5- Diphenyltetrazolium Bromide test [MTT test] was used to determine doses for cell treatment, and the gene expression was analyzed by real-time reverse transcriptase-polymerase chain reaction [real-time RT- PCR]. Significant dose- and time-dependent cell growth inhibitory effects of silybin and silybin-phosphatidylcholine along with ESR1 down-regulation were observed in T47D cells. In contrast to ESR1, the T47D cell line showed negligible ESR2 expression. This study suggests that silybin and silybin-phosphatidylcholine down-regulate ESR1 in ER+ breast cancers. Results also show that in the T47D cell line, silybinphosphatidylcholine has a much higher growth inhibitory effect and a more significant down-regulation of ESR1 compared with silybin