Female ; Humans ; Ovarian Follicle/pathology* ; Ovarian Neoplasms/pathology*

The aim of the present study was to explore the effect and mechanism of Bushen Huoxue recipe (BHR) on ovarian reserve in mice with premature ovarian failure (POF). Mice were divided into 3 groups: normal group, model group and BHR group. Intraperitoneal injection of cyclophosphamide was performed to create the POF model. Primordial follicular (PDF) number, ovarian wet weight, ovarian index, and estrous cycle were analyzed to evaluate the effect of BHR on POF. Meanwhile, the mRNA and protein level of Mouse Vasa Homologue (MVH) in the bone marrow, peripheral blood and ovary were detected, to explore the underlying mechanism of the treatment efficacy of BHR on ovarian reserve. By the time of BHR treatment for 28 days, BHR increased the PDF number and shortened the estrous cycle of POF mice. BHR also decreased the mRNA level of MVH in the bone marrow, and increased mRNA and protein level of MVH in the ovary of POF mice. Our results demonstrated a treatment efficacy of BHR on POF mice, and revealed that BHR might repair the dysfunction of germline stem cells in the bone marrow, and thus to improve the ovarian reserve and enhance the ovarian function of POF mice through neo-oogenesis.
Animals ; Bone Marrow ; drug effects ; metabolism ; Cyclophosphamide ; toxicity ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Estrous Cycle ; drug effects ; Female ; Gene Expression Regulation ; drug effects ; Humans ; Mice ; Ovarian Follicle ; drug effects ; growth & development ; Ovarian Reserve ; drug effects ; Primary Ovarian Insufficiency ; chemically induced ; drug therapy ; pathology

INTRODUCTIONThe aim of this study was to determine the effect of a high-fat diet (HFD) on oocyte maturation and quality in a mouse model.

METHODSFemale BALB/c mice were allocated to one of the following groups: (a) control group (n = 40), which received a controlled diet; or (b) HFD group (n = 40), which received an HFD for 12 weeks. Sections of the ovary were examined histologically. The number of follicles and corpora lutea were counted. In vitro maturation and in vitro fertilisation (IVF) were assessed in germinal vesicle (GV) and metaphase II (MII) oocytes, respectively. The expression of bone morphogenetic protein 15 (BMP15) and leptin receptor genes in GV and MII oocytes was evaluated using reverse transcription real-time polymerase chain reactions.

RESULTSIn the HFD group, there was a decreased number of primordial and Graafian follicles, as well as corpora lutea (p < 0.05). The rate of oocyte development to the MII stage was also reduced (p < 0.001). Cumulus expansion was observed more frequently in the control group than the HFD group (p < 0.05). The IVF rate in the HFD group was lower than that in the control group (p < 0.05). In the HFD group, BMP15 and leptin receptor genes were upregulated in the GV stage (p > 0.05) and MII stage (p < 0.05), compared to the control group.

CONCLUSIONAn HFD reduces folliculogenesis in the primordial and Graafian stages, in vitro maturation and in vitro fertilisation rates, as well as oocyte quality in mice.

Animals ; Body Weight ; Bone Morphogenetic Protein 15 ; metabolism ; Corpus Luteum ; pathology ; Diet, High-Fat ; Female ; Fertility ; Fertilization in Vitro ; methods ; Gene Expression Regulation ; Metaphase ; Mice ; Mice, Inbred BALB C ; Obesity ; complications ; Oocytes ; cytology ; pathology ; Ovarian Follicle ; pathology ; Ovary ; metabolism ; pathology ; Photography ; Polymerase Chain Reaction ; Receptors, Leptin ; metabolism

OBJECTIVE: To investigate whether granulocyte-colony stimulating factor (G-CSF) can decrease the extent of ovarian follicle loss caused by cisplatin treatment. METHODS: Twenty-one adult female Sprague-Dawley rats were used. Fourteen rats were administered 2 mg/kg/day cisplatin by intraperitoneal injection twice per week for five weeks (total of 20 mg/kg). Half of the rats (n=7) were treated with 1 mL/kg/day physiological saline, and the other half (n=7) were treated with 100 microg/kg/day G-CSF. The remaining rats (n=7, control group) received no therapy. The animals were then euthanized, and both ovaries were obtained from all animals, fixed in 10% formalin, and stored at 4degrees C for paraffin sectioning. Blood samples were collected by cardiac puncture and stored at -30degrees C for hormone assays. RESULTS: All follicle counts (primordial, primary, secondary, and tertiary) and serum anti-Mullerian hormone levels were significantly increased in the cisplatin+G-CSF group compared to the cisplatin+physiological saline group. CONCLUSION: G-CSF was beneficial in decreasing the severity of follicle loss in an experimental rat model of cisplatin chemotherapy.
Animals ; Anti-Mullerian Hormone/blood ; Antineoplastic Agents/*toxicity ; Biological Markers/blood ; Cisplatin/*toxicity ; Disease Models, Animal ; Drug Evaluation, Preclinical/methods ; Female ; Fertility Preservation/methods ; Granulocyte Colony-Stimulating Factor/*therapeutic use ; Ovarian Follicle/drug effects/pathology ; Primary Ovarian Insufficiency/blood/chemically induced/pathology/*prevention & control ; Rats, Sprague-Dawley

OBJECTIVETo study the effects of gonadotroph-releasing hormone (GnRH) agonist (GnRH-a) and GnRH antagonist (GnRH-ant) on cyclophosphamide (CTX)-induced follicle apoptosis in female rats.

METHODSThirty-six female Sprague- Dawley rats were randomized into 6 groups, namely normal saline (NS), CTX, GnRH-a+NS, GnRH-a+CTX, GnRH-ant+NS, and GnRH-ant+CTX groups. The rats were sacrificed between the first and second week after the treatments., and the follicle apoptosis was investigated using TUNEL assay and transmission electron microscopy.

RESULTSThe apoptosis rate of the granulose cells in the follicles in late development was significantly higher than that in early follicles, and the apoptosis rate of the oocytes and granulose cells in rats with CTX treatment was significantly higher than that in rats without CTX treatment (P<0.05). The apoptosis rate of the granulose cells in GnRH-a groups (ranging from 33.40 - or + 4.59 to 73.25 - or + 5.35) was significantly higher than that in GnRH-ant groups (27.46 - or + 4.52 to 49.38 - or + 5.02, P<0.05), but there was no significant difference in the oocytes of early follicles between GnRH-a groups (23.48 - or + 4.25 to 36.15 - or + 4.23) and GnRH-ant groups (21.47 - or + 3.81 to 34.04 - or + 5.54, P>0.05). Electron microscopy revealed characteristic apoptotic changes of the oocytes in early follicles and granulose cells in early and late follicles. The apoptotic changes were especially typical in the granulose cells showing the formation of the apoptotic bodies, and the oocytes only showed chromatin condensation and aggregation.

CONCLUSIONIn the rat mode, GnRH-a promotes while GnRH-ant suppressed follicle apoptosis induced by CTX. GnRH analogues regulates mainly granulose cell apoptosis, but have little effect on oocyte apoptosis.

Animals ; Apoptosis ; drug effects ; Cyclophosphamide ; toxicity ; Female ; Gonadotropin-Releasing Hormone ; analogs & derivatives ; antagonists & inhibitors ; Granulosa Cells ; pathology ; Oocytes ; pathology ; Ovarian Follicle ; pathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate expression and clinical significance of VEGF and apoptosis in frozen-thawed mouse ovaries after transplantation.

METHODSOvaries from B6C2F1 (C57BL/6j x BALB/c) 4 week old mice were cryopreservation and the thawed ovaries were xenografted into kidney capsules of 8-12 week old adult male mice. The grafted were recovered 1 d, 2 d and 7 d after transplantation respectively, the grafts and frozen-thawed were removed for follicle counting and immunohistochemically, ultrastructure, and detection of the mRNA expression of vascular endothelial growth factor(VEGF).

RESULTSThe follicle numbers were decreased gradually after transplantation,the cell apoptosis increased especially in 48 h after transplantation. Transmission electron microscopy (TEM) showed the tissue damaged was severest 48 h after transplantation; the expression of VEGF increased after transplantation, peaked on day 7, the mRNA expression of VEGF120 and VEGF188 was more on 48 h after transplantation, decreased on day 7 (P < 0.05).

CONCLUSIONThe number of follicles was decreased after transplantation, the apoptosis index was increased especially in 48 h after transplantation; the expression of VEGF increased after transplantation, an increase in the VEGF188 and VEGF164 isoform might suggest the positive effect in the early stages of angiogenesis.

Animals ; Apoptosis ; physiology ; Cryopreservation ; Female ; Male ; Mice ; Organ Preservation ; adverse effects ; Ovarian Follicle ; pathology ; Ovary ; pathology ; transplantation ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the protective effect of estrogen against cyclophosphamide (CTX)-induced ovarian failure in female rats.

METHODSSixty female Wistar rats (2-3 months old) were randomized into 4 groups to receive treatments with normal saline, CTX, estradiol (E2) or CTX+E2. Serum estradiol and follicle-stimulating hormone (FSH) concentrations were measured regularly in the 4 groups, and the weight of the ovary and uterus, follicle number and mean diameter of the follicles were compared.

RESULTSCompared with CTX+EE2 group, the CTX group showed significantly increased FSH levels and decreased EE2 levels (P<0.05). The weight of the ovary and uterus and follicle number were significantly lower in CTX group than in CTX+EE2 group (P<0.05). No obvious differences in the indexes were found between the control and CTX+E2 groups.

CONCLUSIONEstrogen administration provides protection against CTX-induced ovarian damage in rats.

Animals ; Cyclophosphamide ; adverse effects ; Estradiol ; blood ; therapeutic use ; Female ; Follicle Stimulating Hormone ; blood ; Ovary ; drug effects ; pathology ; Primary Ovarian Insufficiency ; chemically induced ; prevention & control ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVEWe dynamically measured serum inhibin B and estradiol in the early stage of hormonal stimulation to predict the ovarian response in in vitro fertilization (IVF) treatment.

METHODSA total of 57 patients (<40 years of age) who underwent the first cycle of long protocol IVF or introcytoplasmic sperm injection (ICSI) treatment were included. Serum inhibin B, estradiol, follicle stimulating hormone (FSH) and luteinizing hormone (LH) levels were measured four times: (1) on Day 3 of the menstrual cycle (basal); (2) on the day before the first administration of gonadotrophin (Gn) (Day 0); (3) on Day 1 of Gn therapy; and (4) on Day 5 of Gn therapy. Comparisons of these measurements with ovarian responses and pregnancy outcomes were made and analyzed statistically.

RESULTS(1) On Day 1 and Day 5 of recombinant FSH (rFSH) stimulation, ovarian response, i.e., numbers of follicles, oocytes, fertilized oocytes, and embryos, had a positive correlation (r(s)=0.46~0.61, P=0.000) with raised inhibin B and estradiol concentrations, but a negative correlation (r(s)=-0.67~-0.38, P=0.000 or P<0.01) with total rFSH dose and total days of rFSH stimulation. (2) No significant variation (P>0.05) between the pregnant and non-pregnant groups on the basis of mean age or on all hormone concentrations at four times of the IVF cycle was observed. However, all the seven patients aged >35 years did not reach pregnancy.

CONCLUSIONS(1) Serum inhibin B and estradiol concentrations obtained shortly after Gn therapy may offer an accurate and early prediction of ovarian response; (2) Low levels of serum inhibin B and estradiol obtained shortly after Gn stimulation indicate the need for a longer period of Gn treatment and a higher daily dosage; (3) No obvious pregnancy difference among patients of age <35 years was found; however, IVF pregnancy outcome is significantly lower in women of age >35 years.

Adult ; Cell Count ; Estradiol ; blood ; Female ; Fertilization in Vitro ; Follicle Stimulating Hormone ; administration & dosage ; therapeutic use ; Humans ; Infertility, Female ; blood ; pathology ; therapy ; Inhibins ; blood ; Menstrual Cycle ; Oocytes ; Ovarian Follicle ; drug effects ; physiology ; Ovary ; drug effects ; physiology ; Ovulation Induction ; methods ; Prognosis ; Treatment Outcome

OBJECTIVETo probe the effect of Zuogui pill (ZGP), a Chinese compound recipe for tonifying Shen, on ovarian function in mice with premature ovarian failure (POF).

METHODSBALB/C female mice model of POF was established by multiple sites subcutaneous injection of ovarian antigen elicited with ovarian tissue of SD female rats, and treated with ZGP at different time points in the modeling, with prednisone as positive control. The levels of follicle-stimulating hormone (FSH) and estradiol (E2) in peripheral blood were measured with radioimmunoassay, and ovarian antibody (AoAb) was determined by enzyme linked immunosorbent assay. The mRNA expression of ovarian growth and differentiation factor-9 was detected with in situ hybridization.

RESULTSPOF model mice manifested such abnormalities as increased FSH, decreased E2, and positive AoAb in peripheral blood, with lymphocytes infiltration in ovarian mesanchyma, reduction of GDF-9 mRNA positive oocytes, and decrease of growing and mature follicles. ZGP could reduce the increase of FSH, increase the level of E2, inhibit the production of AoAb, raise the GDF-9 mRNA positive cells of oocytes, increase the number of growing and mature follicles. The clinical efficacy was more significant in early stage than in advanced stage.

CONCLUSIONZGP can improve immune inflammatory injury of ovary, and shows therapeutic effect on POF.

Animals ; Autoimmune Diseases ; etiology ; prevention & control ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred BALB C ; Ovarian Follicle ; pathology ; Ovary ; drug effects ; immunology ; physiopathology ; Phytotherapy ; Primary Ovarian Insufficiency ; etiology ; prevention & control ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Yin Deficiency ; drug therapy

OBJECTIVETo investigate whether the follicle stimulating hormone (FSH) can inhibit apoptosis in ovarian cancer cells induced by cisplatin (DDP) and its possible mechinism.

METHODSDNA fragmentation assay, (TdT-mediated dUTP nick end labling TUNEL), Western blot were used to analyze the changes in expression levels of Survivin and bcl-2 protein. The relative activity of caspase-3 was also determined.

RESULTS200 mIU/ml FSH could regulate down the percentage of apoptotic cells and DNA fragmentation induced by 5.0 micrograms/ml cisplatin, while 200 mIU/ml FSH increased Survivin protein expression but could't influence the expression of bcl-2 protein.

CONCLUSIONFSH can inhibit ovarian cancer cells apoptosis induced by cisplatin. The possible mechinism is up-regulation of Survivin expression and down-regulation of caspase activity.

Antineoplastic Agents ; antagonists & inhibitors ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; Caspases ; metabolism ; Cisplatin ; antagonists & inhibitors ; pharmacology ; DNA Fragmentation ; Female ; Flow Cytometry ; Follicle Stimulating Hormone ; pharmacology ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; metabolism ; Neoplasm Proteins ; Ovarian Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Receptors, FSH ; metabolism ; Tumor Cells, Cultured