OBJECTIVE: To assess the association of gene expression with development potential of early embryos derived from patients with polycystic ovary syndrome (PCOS). METHODS: Three pairs of infertile patients with respectively matched age, body mass index, ovarian reserve and treatment with gonadotrophin-releasing hormone (GnRH) antagonists were selected. Patients with fewer embryos were assigned as the case group (n = 3), whilst the remainders were assigned as the control group (n = 3). Ovarian granulosa cells from patients were collected for the extraction of total RNA and subjected to RNA sequencing. The results were subjected to differential gene expression and functional enrichment analyses. RESULTS: Compared with the control group, 76 genes were up-regulated and 110 genes were down-regulated in the case group. The level of estradiol (E₂) was significantly higher in the control group on the trigger day with the injection of human chorionic gonadotrophin (HCG). Compared with the control group, the KRT7 gene was most significantly up-regulated, whilst the CCNYL2 gene was most significantly down-regulated in the case group. Gene ontology (GO) entries enrichment has found those associated with chromosome segregation, cell cycle regulation, and fatty acid metabolism to be significantly enriched. The genes participating in the regulation of cell assembly, differentiation, negative regulation of cell cycle, negative regulation of development, extracellular regulated protein kinases (ERK), ERK1 and ERK2 signaling pathways to be significantly down-regulated. KEGG enrichment analysis of cell signaling pathways revealed that steroid hormone biosynthesis-related genes were enriched. CONCLUSION Among patients treated with GnRH antagonists, the significant difference in the number of oocytes fertilized in vitro and the number of available embryos are associated with the difference in the expression of genes of ovarian granulosa cells.
Female ; Humans ; Pregnancy ; Embryonic Development ; Gene Expression ; Gonadotropin-Releasing Hormone/antagonists & inhibitors* ; Granulosa Cells ; Polycystic Ovary Syndrome/genetics*

OBJECTIVE: To screen proteins interacting with ring finger protein 216(RNF216) through yeast two hybrid experiment, and further clarify the role of RNF216 in the pathogenesis of gonadotropin-releasing hormone deficiency. METHODS: A recombinant expression vector pGBKT7-RNF216 was constructed and transformed into yeast Y2HGold, which was hybridized with a human cDNA library in order to screen proteins interacting with RNF216. The interaction was verified in yeast Y2HGold. RESULTS: A recombinant expression vector pGBKT7-RNF216 was successfully constructed and expressed in yeast Y2HGold. Filamin B (FLNB) was identified by yeast two hybrid experiment, and their interaction was verified in yeast Y2HGold. CONCLUSION An interaction between FLNB and RNF216 was identified through yeast two hybrid experiment. RNF216 may affect the proliferation and migration of GnRH neurons by regulating FLNB or FLNB/FLNA heterodimers.
Gene Library ; Gonadotropin-Releasing Hormone/genetics* ; Humans ; Proteins ; Two-Hybrid System Techniques ; Ubiquitin-Protein Ligases/genetics*

The transcription factor SOX10, as a major actor in the development of the neural crest, plays a key role in the maintenance of progenitor cell multipotency, lineage specification, and cell differentiation. Abnormalities of neural crest development in humans lead to a number of genetic diseases known as neurocristopathies or neural crest disorders. The mutation of SOX10 can cause Kallmann syndrome (KS), which is a clinically and genetically heterogeneous condition and defined by the association between anosmia and hypogonadotropic hypogonadism due to incomplete migration of neuroendocrine gonadotropin-releasing hormone (GnRH) cells along the olfactory, vomeronasal, and terminal nerves. Since then, there have been a number of related reports that mutation of SOX10 will lead to KS with deafness. This review focuses on the SOX10 gene and the advances in the diagnosis and genetic studies of KS with deafness caused by the mutatuin of SOX10.
Cell Differentiation ; Deafness ; genetics ; Gonadotropin-Releasing Hormone ; Humans ; Hypogonadism ; Kallmann Syndrome ; genetics ; Mutation ; genetics ; SOXE Transcription Factors ; genetics

OBJECTIVETo observe the effect of nourishing yin removing fire Chinese herbs (NYRF-CH) on the gene expression of hypothalamic growth hormone secretion peptide (Ghrelin) and its receptor growth hormone secretion peptide receptor 1alpha (GHSR1-alpha) at the puberty onset of danazol induced female precocious rats.

METHODSForty female SD rats were randomly divided into 4 groups, i.e., the normal group (N), the model group (M), the normal saline intervention group (NS), and the NYRFCH intervention group (NI), 10 in each group. 300 microg danazol was subcutaneously injected to all rats except those in the N group to prepare precocious rat model. NYRFCH and normal saline was respectively administered to rats in the NI and the NS group from the 15th day old for 7-10 days. No treatment was given to rats in the N group. Time of rats' vulva opening was recorded. Ovary index and uterus index were calculated. Peripheral blood levels of estradiol (E2), follicle stimulating hormone (FSH), and luteinizing hormone (LH), and hypothalamic contents of gonadotropin releasing hormone (GnRH) as well as the gene expression of hypothalamic Ghrelin and GHSR1-alpha were determined. Results Compared with the N group, the vulva opening time was advanced in the model group; peripheral blood levels of E2 and LH, uterus index, hypothalamic contents of GnRH increased; peripheral blood FSH levels and mRNA levels of hypothalamic Ghrelin and GHSR1-alpha decreased (P < 0.05, P < 0.01). Compared with the M group and the NS group, the vulva opening time was not advanced in the NI group; peripheral blood levels of E2 and LH, uterus index and hypothalamic contents of GnRH obviously decreased (P < 0.05, P < 0.01); mRNA levels of hypothalamic Ghrelin and GHSR1-alpha increased (all P < 0.01). But there was no statistical difference in the hypothalamic contents of Ghrelin, or the number and activity of GHSR1-alpha (P > 0.05).

CONCLUSIONNYRFCH had regulatory effect on regulating hypothalamic Ghrelin and GHSR1-alpha at gene transcription levels.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Estradiol ; Female ; Follicle Stimulating Hormone ; metabolism ; Gene Expression ; drug effects ; Ghrelin ; genetics ; Gonadotropin-Releasing Hormone ; metabolism ; Hypothalamus ; metabolism ; Luteinizing Hormone ; metabolism ; Ovary ; Puberty, Precocious ; metabolism ; Rats ; Rats, Sprague-Dawley ; Uterus

The effects of fluoride exposure on the functions of reproductive and endocrine systems have attracted widespread attention in academic circle nowadays. However, it is unclear whether the gene-environment interaction may modify the secretion and activity of hypothalamus-pituitary- ovarian (HPO) axis hormones. Thus, the aim of this study was to explore the influence of fluoride exposure and follicle stimulating hormone receptor (FSHR) gene polymorphism on reproductive hormones in Chinese women. A cross sectional study was conducted in seven villages of Henan Province, China during 2010-2011. A total of 679 women aged 18-48 years were recruited through cluster sampling and divided into three groups, i.e. endemic fluorosis group (EFG), defluoridation project group (DFPG), and control group (CG) based on the local fluoride concentration in drinking water. The serum levels of gonadotropin releasing hormone (GnRH), follicle stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) were determined respectively and the FSHR polymorphism was detected by real time PCR assay. The results provided the preliminary evidence indicating the gene-environment interaction on HPO axis hormones in women.
Adolescent ; Adult ; Age Factors ; Asian Continental Ancestry Group ; China ; Cross-Sectional Studies ; Estradiol ; blood ; Female ; Fluoridation ; adverse effects ; Fluorides ; administration & dosage ; adverse effects ; urine ; Follicle Stimulating Hormone ; blood ; Gene-Environment Interaction ; Gonadotropin-Releasing Hormone ; blood ; Humans ; Hypothalamus ; physiology ; Luteinizing Hormone ; blood ; Middle Aged ; Ovary ; physiology ; Pituitary Gland ; physiology ; Polymorphism, Single Nucleotide ; Receptors, FSH ; genetics ; Tobacco Smoke Pollution ; Young Adult

OBJECTIVETo study the therapeutic effect of Dabuyin Wan on true precocious puberty of female rats and its possible mechanism.

METHODTwenty-two-day-old female SD rats were subcutaneously injected with 40 mg x kg(-1) N-methyl-DL-aspartic acid (NMA) at 14:00 and 16:00 every day; meanwhile, the rats were given Dabuyin Wan for intervention. Visual inspection was conducted for the time of vaginal opening. The first estrus was observed by yaginal smear test. Their ovaries and uterus were weighed to calculate organ coefficients. Conventional pathological slices were made to observe morphological changes in ovaries and uterus and calculate the thickness of uterine walls and the number of corpus luteums. The level of E2 in serum was detected to assess the therapeutic effect of Dabuyin Wan on NMA precocious puberty in rats. expressions of GnRH, GPR54 and Kiss-1 mRNA in hypothalamus were measured by semi-quantitative RT-PCR to investigate the possible mechanism of Dabuyin Wan.

RESULTDabuyin Wan at 3.24 g x kg(-1) and 1.62 g x kg(-1) significantly decreased the organ coefficients in rats with precocious puberty (P < 0.05), decrease the number of vaginal openings in rats (P < 0.01) and the thickness of uterine walls and the number of corpus luteums (P < 0.05), and notably down-regulated expressions of GnRH, GPR54 and Kiss-1 mRNA in hypothalamus (P < 0.05), without significant impact on E2 in serum.

CONCLUSIONDabuyin Wan may inhibit GnRH synthesis and release as well as startup of hypothalamic-pituitary-gonadal axis by down-regulating Kiss-1/GPR54 mRNA expression in hypothalamus, in order to realize the therapeutic effect on true precocious puberty.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Estrus ; drug effects ; Female ; Gene Expression ; drug effects ; Gonadotropin-Releasing Hormone ; genetics ; Hypothalamus ; drug effects ; metabolism ; Kisspeptins ; genetics ; Ovary ; drug effects ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, G-Protein-Coupled ; genetics ; Receptors, Kisspeptin-1 ; Reverse Transcriptase Polymerase Chain Reaction ; Sexual Maturation ; drug effects ; genetics ; Time Factors ; Uterus ; drug effects ; Vagina ; drug effects

This study is to investigate the anti-tumor effect in vitro of methotrexate modified by LH-RH peptide (LH-RH-MTX). LH-RH receptors highly expressing MCF-7 human breast carcinoma cell line and lowly expressing K562 human erythroleukemia cell line were served as the tested cells. The cell proliferation inhibition rates of LH-RH-MTX were detected by MTT colorimetric assay. The effects of LH-RH-MTX on the cell cycle and apoptosis rates were detected by flow cytometry. The inhibition rate of LH-RH-MTX on MCF-7 cells was much higher than that on K562 cells, and the inhibition rate of LH-RH-MTX on MCF-7 cells was much higher than that of free MTX at the same concentration. The inhibition rate of LH-RH-MTX on rat bone marrow mononuclear cells was less than that of free MTX. The number of MCF-7 cells in S phase increased after administration of LH-RH-MTX. The apoptosis rate of LH-RH-MTX group significantly increased compared with that of the control group and MTX group. The relative expression of LHRHR mRNA of LH-RH-MTX group markedly decreased compared with that of the control group and MTX group. LH-RH-MTX is realizable to reduce drug side effects, increase the therapeutic index and achieve tumor-targeted therapy.
Animals ; Antimetabolites, Antineoplastic ; chemical synthesis ; pharmacology ; Apoptosis ; drug effects ; Bone Marrow Cells ; cytology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drug Delivery Systems ; Gonadotropin-Releasing Hormone ; chemistry ; pharmacology ; Humans ; K562 Cells ; Leukocytes, Mononuclear ; MCF-7 Cells ; Methotrexate ; chemical synthesis ; pharmacology ; RNA, Messenger ; metabolism ; Rats ; Receptors, LHRH ; biosynthesis ; genetics

BACKGROUNDType I gonadotropin-releasing hormone (GnRH-I) agonists have been applied for the treatment of steroid-dependent tumors such as breast carcinoma, ovarian cancer and prostatic carcinoma. But the mechanism has not been clarified yet. There are few reports about the treatment of endometrial carcinoma using GnRH-I agonists. Type II GnRH (GnRH-II) is a new subtype of GnRH. Our aim was to investigate the effects of GnRH-I agonists and GnRH-II on estrogen receptor-negative human endometrial carcinoma cells and the effect from phosphatase and tensin homolog gene (PTEN) to them.

METHODSA lentiviral vector-mediated RNAi method was used to establish a PTEN-negative HEC-1A cell clone (HEC-1A-ND). MTT and flow cytometry were used to detect the cell proliferation, cell cycle and apoptosis of HEC-1A, HEC-1A-NC and HEC-1A-ND cells after treatment with GnRH-I agonist Triptorelin (10(-11) mol/L to 10(-5) mol/L) or GnRH-II (10(-11) mol/L to 10(-5) mol/L). Western blotting was used to detect AKT and ERK1/2 activation after treatment with different concentrations of Triptorelin or GnRH-II for 30 minutes in the above mentioned three kinds of cells.

RESULTSTriptorelin and GnRH-II induced apoptosis and inhibited proliferation of HEC-1A, HEC-1A-ND and HEC-1A-NC in a dose-dependent manner. This effect was augmented in HEC-1A-ND cells in which PTEN gene was knocked-down. Furthermore, Triptorelin and GnRH-II inhibited the AKT and ERK activity in HEC-1A-ND cells.

CONCLUSIONSTriptorelin and GnRH-II can promote apoptosis rate and inhibit cell proliferation of estrogen receptor-negative endometrial carcinoma cells in a dose-dependent manner. PTEN gene can inhibit the effects of Triptorelin or GnRH-II on human endometrial carcinoma cells.

Apoptosis ; drug effects ; Blotting, Western ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Endometrial Neoplasms ; genetics ; metabolism ; Female ; Gonadotropin-Releasing Hormone ; analogs & derivatives ; pharmacology ; Humans ; PTEN Phosphohydrolase ; genetics ; physiology ; RNA Interference ; Triptorelin Pamoate ; pharmacology

AIMTo study the effect and mechanism of gonadotrophin-releasing hormone (GnRH) on murine Leydig cell steroidogenesis.

METHODSPurified murine Leydig cells were treated with GnRH-I and -II agonists, and testosterone production and steroidogenic enzyme expressions were determined.

RESULTSGnRH-I and -II agonists significantly stimulated murine Leydig cell steroidogenesis 60%-80% in a dose- and time-dependent manner (P < 0.05). The mRNA expressions of steroidogenic acute regulatory (StAR) protein, P450scc, 3beta-hydroxysteroid dehydrogenase (HSD), but not 17alpha-hydroxylase or 17beta-HSD, were significantly stimulated by both GnRH agonists with a 1.5- to 3-fold increase (P < 0.05). However, only 3beta-HSD protein expression was induced by both GnRH agonists, with a 1.6- to 2-fold increase (P < 0.05).

CONCLUSIONGnRH directly stimulated murine Leydig cell steroidogenesis by activating 3b-HSD enzyme expression.

3-Hydroxysteroid Dehydrogenases ; biosynthesis ; genetics ; Animals ; Blotting, Western ; Cell Separation ; Cells, Cultured ; Cholesterol Side-Chain Cleavage Enzyme ; biosynthesis ; Dose-Response Relationship, Drug ; Gonadotropin-Releasing Hormone ; agonists ; pharmacology ; Leydig Cells ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Phosphoproteins ; biosynthesis ; genetics ; RNA ; biosynthesis ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sexual Maturation ; physiology ; Steroids ; biosynthesis ; Testosterone ; biosynthesis

BACKGROUNDOvarian hyperstimulation syndrome (OHSS) is one of the most life-threatening complications of assisted reproduction treatments. Gonadotropin-releasing hormone antagonists (GnRHanta) are thought to be effective in preventing this complication, and some clinical trials have found lower incidences of OHSS in patients treated with GnRHanta. Our aim was to investigate the effects of GnRHanta on vascular permeability and the expression of vascular endothelial growth factor (VEGF) and its receptors in a rat model of OHSS.

METHODSAn immature early OHSS rat model was established. Three ovarian stimulation protocols were used: pregnant mare serum gonadotropin/human chorionic gonadotropin (hCG) alone, with a GnRHanta, or with a gonadotropin-releasing hormone agonists (GnRHa). Blood and tissue samples were collected at 48 hours after hCG administration. Vascular permeability was evaluated by measuring the Evans-Blue content of extravasated peritoneal fluids. The expression of VEGF and its receptors, including flt-1 and KDR, were detected by reverse transcriptase-polymerase chain reaction and Western blotting.

RESULTSTreatment with both a GnRHanta and a GnRHa resulted in significant reductions in serum estradiol and peritoneal vascular permeability, as well as decreased ovarian expression of VEGF and its two receptors. However, GnRHanta treatment caused a greater reduction in serum estradiol concentrations, and in VEGF receptor mRNA expression than GnRHa. There were no significant reductions in the expression of VEGF or its receptors in extra-ovarian tissues, including the liver, lungs and peritoneum.

CONCLUSIONOur results reveal that GnRHanta are more potent than GnRHa in preventing early OHSS through down-regulation of the expression of VEGF and its receptors in hyperstimulated ovaries.

Animals ; Blotting, Western ; Chorionic Gonadotropin ; pharmacology ; therapeutic use ; Disease Models, Animal ; Female ; Gene Expression ; drug effects ; Gonadotropin-Releasing Hormone ; analogs & derivatives ; antagonists & inhibitors ; pharmacology ; therapeutic use ; Ovarian Hyperstimulation Syndrome ; drug therapy ; genetics ; metabolism ; Rats ; Rats, Wistar ; Receptors, Vascular Endothelial Growth Factor ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; genetics ; metabolism