Effects of Anethum graveolens L. on in vitro matured mouse oocytes and granulosa cells

Background: According to previous studies, Anethum graveolens L. [dill] aqueous extracts decreased the fertility of female rats. Therefore, the present study aimed to examine the effects of this herb on cultured granulosa cells and immature oocytes

Methods: The cells were obtained from 27-29 day immature superovulated mice. The oocytes were cultured in a petri dish consisting of 30 micro l drops of MEM-alpha and granulosa cells in a 24-well plate consisting of DMEM/F12 and different concentrations of 0, 10, 50, 100, 500, 1000, 10000 microg/ml of dill seed aqueous extract [DSAE] in 37oC and 5% CO2. Then, the in vitro maturation of oocytes, including Germinal Vesicle [GV], Germinal Vesicle Breakdown [GVBD], and meiosis ?? [M??] and oocyte bioviability were determined. Granulosa cells were then extracted and their bioviability, apoptosis, chromatin condensation, and lipid synthesis were examined. Estrogen and progesterone concentrations and Alkaline Phosphatase [ALP] activity were measured by RIA and spectrophotometry respectively from the supernatant of granulosa cell culture

Results: The results revealed that concentration of 10000 microg/ml of DSAE were toxic and damaged granulosa cell growth and oocytes maturation. Lower concentrations were the same in the control group and did not have any side effects on cell growth. The number of lipid droplets, estrogen and progesterone concentrations, and ALP activity increased with higher doses of DSAE compared to those in the control culture. Additionally, apoptosis and chromatin condensation increased in higher concentrations of DSAE-[500 and 1000 micro g/ml] treated cells. This herb extract decreased the oocytes maturation in dose-dependent manner

Conclusion: It was concluded that DSAE increased granulosa cells activity but damaged oocytes maturation, therefore it might be introduced as infertility agent

effects of Salvia officinalis L. on granulosa cells and in vitro maturation of oocytes in mice

Background: Salvia officinalis L. has been used since ancient times but there are little data about effects of this herb on normal reproductive cells

Objective: To investigate the toxicity effects of Salvia officinalis L. on granulosa cells [GCs] and maturation of oocytes

Materials and Methods: GCs and oocytes were extracted from superovulated ovaries of immature mice. The cells were treated with concentrations of 10, 50, 100, 500, and 1000 microg/ml of Salvia officinalis hydroalcoholic extracts and compared with the control culture. Bioviability, chromatin condensation, estradiol and progesterone concentrations, lipid synthesis, apoptosis, and alkaline phosphatase activity of GCs were measured. In vitro maturation of oocytes by determination of different maturation stages of oocytes including germinal vesicle, germinal vesicle breaks down, and metaphase II were examined

Results: The results revealed that 500 and 1000 microg/ml concentrations of Salvia officinalis L. were toxic. The most of the GCs were in the early stages of apoptosis in 100 microg/ml treated culture and cell death happened with 500 microg/ml treatment. Progesterone concentration was reduced in 100 microg/ml and higher doses but estradiol concentration and alkaline phosphatase showed opposite effects. The lipid droplets content of GCs reduced significantly in all groups especially in 500 and 1000 microg/ml. Finally, oocyte's nucleus and cytoplasm showed a high level of condensation, and meiosis rate reduced in all treated cultures

Conclusion: Our findings suggested that higher dose of Salvia officinalis hydroalcoholic extracts inhibits, oocyte maturation, GCs bioviability, proliferation, and secretion

Effects of Anethum graveolens L. [dill] on oocyte and fertility of adult female rats

Our previous studies revealed Anethum graveolens L. caused some changes in female reproductive system that induced infertility. Therefore, in this study, oocyte changes as one of probable reasons of infertility were investigated. In this study, 59 adult female rats were divided into 3 groups of control, low dose [0.5 g/kg] and high dose [5 g/kg] of dill seed aqueous extract [LDE and HDE] treated groups that were gavaged with 1 ml of each dose for 10 days [2 estrous cycles]. Vaginal smears were prepared daily. Oocytes of superovulated animals were extracted and their morphometrical changes were measured [n=5]. Oocyte cell membrane glycoconjugates were stained with UEA, PNA, and DBA-FITC lectins [n=5]. Ultrastructural studies of oocytes were performed using TEM [n=5]. The number, weight, and crown-rump length of newborns were examined in three groups after mating with untreated males [n=5]. Data were analyzed using SPSS software. Results demonstrated that the duration of the estrous cycle, the diestrus phase and progesterone concentration in the experimental groups increased significantly compared to the control group [p<0.05]. Granulosa cells of corpus luteum in HDE-treated group were larger and clearer. The intensity reactions of galactose/N-acetylgalactoseamine terminal sugar of oocyte decreased insignificantly in experimental groups compared to the control group p>0.05. Duration of mating to pregnancy increased and the weight and crown-rump length of newborns decreased in experimental groups significantly [p<0.05]. Dill seed aqueous extract can induce infertility without any effect on oocyte structure

Effect of pomegranate juice on bone calcium content and body weight of adult mice

Pomegranate juice has several antioxidant components such as flavenoids and mineral materials such as sodium and potassium. In this study the effects of pomegranate juice on bone calcium content and body weight of adult mice were survived. In this applied study two doses [3.3 and 6.6 ml/kg] of pomegranate juice [PJ] were gavaged to female mice for 30 days. Animals were weighed at days of 0, 5, 10, 15, 20, 25 and 30. Bone calcium contents were measured by flame photometer. Bone calcium content of PJE treated mice increased but it was not significant statistically. Pomegranate juice did not affect body weight. Pomegranate juice extracts even its high dose did not show any side effect on body weight and tissues of adult female mice

Mesenchymal stem cells repair germinal cells of seminiferous tubules of sterile rats

Mesenchymal stem cells [MSCs] are undifferentiated cells that can differentiate and divide to other cell types. Transplantation of these cells to the different organs is used for curing various diseases. The aim of this research was whether MSCs transplantation could treat the sterile testes. In this experimental study, Donor MSCs were isolated from bone marrow of Wistar rats. The recipients were received 40 mg/kg of busulfan to stop endogenous spermatogenesis. The MSCs were injected into the left testes. Cell tracing was done by labeling the MSCs by 5-Bromo-2- Deoxy Uridine [BrdU]. The immunohistochemical and morphometrical studies were performed to analysis the curing criteria. The number of spermatogonia [25.38 +/- 1.57], primary spermatocytes [55.41 +/- 1.62] and spermatozoids [4.95 +/- 1.30]x10[6] in busulfan treated animals were decreased significantly as compared to the control group [33.35 +/- 1.78, 64.44 +/- 2.00] and [10.50 +/- 1.82]x10[6] respectively but stem cells therapy help the spermatogenesis begin more effective in these animals [32.78 +/- 1.99, 63.59 +/- 2.01] and [9.81 +/- 1.33]x10[6] respectively than the control group. The injected BrdU labeled mesenchymal stem cells differentiated to spermatogonia and spermatozoa in the seminiferous tubules of the infertile testis and also to the interstitial cells between tubules. We concluded that testis of host infertile rats accepted transplanted MSCs. The transplanted MSCs could differentiate into germinal cells in testicular seminiferous tubules

Effects of plum extract on skeletal system of fetal and newborn mice

To evaluate the effects of Prunus domestica L. extracts on fetuses and neonatal skeletal systems. A total of 32 pregnant mice [Mus musculus] received vehicle and plum hydroalcoholic extract at gestational days 1-18 and during the entire gestational period as well as 10 days postpartum, respectively. A total of 30 nonpregnant mice were fed plum hydroalcoholic extract and plum juice extract for 30 days. Bone calcium content and serum concentrations of calcium, magnesium and alkaline phosphatase were measured. The skeletal systems of their fetuses and neonates were stained with Alcian blue and alizarin red S and the length of femur, tibia, and their ossification center were measured. Crown-rump length of the newborn mice from mothers treated with plum extract [4.61 +/- 0.25 mm] was higher compared to the control group [4.48 +/- 0.31 mm, p = 0.001], and the femur osteogenesis index of newborn mice from mothers treated with plum extract was also higher [0.87 +/- 0.09] compared to the control group [0.81 +/- 0.06, p = 0.007]. The findings showed that pregnant mice treated with plum extract had fetuses and newborn mice with higher osteogenesis index than those of the controls