Extracellular vesicles (EVs) are membrane-bound particles actively released by cells. In prokaryotes and eukaryotes, EVs are effective bridges for communication between cells. EVs carry biological macromolecules, including proteins, lipids and nucleic acid, which affects different physiological functions of parent cells and recipient cells. Among them, the microRNA carried by EVs is the most reported and plays an important role in physiological function of organisms. During the development of follicles, only a few follicles can fully develop and ovulate, whereas most of them undergo atresia at different stages of development. In the whole process of follicular development, the changes at each stage and the regulation mechanism of follicular atresia are not completely understood. In this paper, we introduced the types, characteristics, isolation methods and uses of EVs, and emphasized how microRNA carried by EVs in follicular fluid regulated follicular atresia from the aspects of different cytokines and hormones. Additionally, the application prospect of microRNA carried by EVs in follicular fluid in reproductive regulation and reproductive disease diagnosis was discussed. This paper is significant for studying the regulation of follicular development and the effective utilization of oocytes.
Animals ; Extracellular Vesicles/metabolism* ; Female ; Follicular Atresia ; Follicular Fluid ; MicroRNAs/metabolism* ; Oocytes

BACKGROUNDThe mechanisms of endometriosis with infertility have not been fully studied. The present study aimed to assess the follicular fluid (FF) levels of prostaglandin E2 (PGE2), which plays a critical role within the ovary, and to investigate the effect of PGE2 on steroidogenesis in granulosa-lutein cells (GLCs) from women with and without endometriosis.

METHODSThirty-three women with laparoscopically documented endometriosis and 40 controls undergoing in vitro fertilization (IVF) were studied. We assayed the concentrations of PGE2 in FF, the production of E2 and progesterone in FF and in culture medium, and the expression of steroidogenic acute regulatory protein (StAR) and CYP19A1 in GLCs with the intervention of PGE2.

RESULTSPGE2 and progesterone concentrations were increased and displayed positive correlation in endometriotic FF. PGE2 induced the expression of StAR and the production of progesterone in GLCs from women with endometriosis, and the expression of StAR and the production of progesterone were increased in GLCs from women with endometriosis. However, there were no significant effects of PGE2 on promoting the production of E2 or the expression of CYP19A1 in GLCs. Moreover, the production of E2 and the expression of CYP19A1 in GLCs from women with endometriosis were significantly decreased compared to the controls.

CONCLUSIONSPGE2 concentrations are increased in endometriotic FF, along with concomitant increases in progesterone and StAR. In contrast, the E2 and CYP19A1 are decreased in GLCs, which may delay the development of the follicles and cause an imbalance in the follicular steroid hormone levels. These changes may have close relationship with endometriosis-associated infertility.

Adult ; Dinoprostone ; metabolism ; Embryo Transfer ; Endometriosis ; metabolism ; Female ; Fertilization in Vitro ; Follicular Fluid ; metabolism ; Humans ; Luteal Cells ; Pregnancy

This prospective study investigated the relationship between anti-Mullerian hormone (AMH) level in the follicular fluid (FF) and the quality of the oocyte and embryo. A total of 65 FF samples from 54 women were included in this study. FF was collected from the largest preovulatory follicle sized> or =20 mm of mean diameter from each ovary. Samples were divided into 3 groups according to the FF AMH levels: below the 33th percentile (low group, FF AMH<2.1 ng/mL, n=21), between the 33th and the 67th percentile (intermediate group, FF AMH=2.1-3.6 ng/mL, n=22), and above the 67th percentile (high group, FF AMH>3.6 ng/mL, n=22). The quality of the ensuing oocytes and embryos was evaluated by fertilization rate and embryo score. FF AMH levels correlated positively with the matched embryo score on day 3 after fertilization (r=0.331, P=0.015). The normal fertilization rate was significantly lower in the low group than in the intermediate group (61.9% vs. 95.5% vs. 77.3%, respectively, P=0.028). Our results suggest that the FF AMH level could be a predictor of the ensuing oocyte and embryo quality.
Adult ; Anti-Mullerian Hormone/*analysis ; Embryo, Mammalian/*cytology ; Female ; Fertilization in Vitro ; Follicular Fluid/*metabolism ; Humans ; Oocytes/cytology ; Prospective Studies

Extracellular vesicles (EVs) refer to bilayer membrane transport vesicles secreted by cells. EVs can take macromolecules from cells and transfer them to receptor cells. Among these macromolecular substances, the most studied are microRNAs (miRNAs). miRNA is non-coding RNA involved in the regulation of gene expression. It has been confirmed that there are different non-coding RNAs in mammalian follicular fluid EVs. EVs carrying miRNA can act as an alternative mechanism for autocrine and paracrine, affecting follicular development. This paper systematically introduced the kinds, characteristics and methods of isolation and identification of EVs, focusing on the effects of EVs and miRNAs on follicular development, including early follicular development, oocyte maturation, follicular dominance and effects on granulosa cell function. At the same time, the authors prospected the future research of EVs and microRNAs in follicular fluid, and provided ideas and directions for the research and application of EVs and miRNA functions in follicular fluid.
Animals ; Extracellular Vesicles ; metabolism ; Female ; Follicular Fluid ; chemistry ; Granulosa Cells ; drug effects ; MicroRNAs ; pharmacology ; Oogenesis ; drug effects

OBJECTIVETo explore the relationship between protein oxidation levels in the follicular fluid and the outcome parameters of in vitro fertilization-embryo transplantation (IVF-ET).

METHODSThe levels of advanced oxidation protein products (AOPP) in the follicular fluid were measured in 64 women with tubal infertility undergoing IVF-ET. The relationship between the AOPP levels and IVF-ET outcome parameters was analyzed.

RESULTSAOPP levels showed significant inverse correlations between the proportion of mature oocytes (r=-0.401, P=0.001), fertilization rate (r=-0.257, P=0.045), cleavage rate (r=-0.290, P=0.024) and good embryo rate (r=-0.520, P=0.000). AOPP levels differed significantly between the groups with different retrieved oocyte numbers (F=3.851, P=0.027), being the lowest in women with 8 to 15 retrieved oocytes and the highest in those with retrieved oocytes below 8. The AOPP level in the non-pregnant women was significantly higher than that in the pregnant women (t=3.665, P=0.001). The AOPP levels also differed significantly with age (F=15.919, P=0.000), and the women >35 years of age had the highest level and those below 30 years had the lowest level.

CONCLUSIONProtein oxidative stress is present in the follicular fluid of women on IVF-ET cycles. High level of AOPP may have adverse effects on the oocytes and early embryonic development and may affect the outcome of IVF-ET.

Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; Follicular Fluid ; metabolism ; Humans ; Infertility, Female ; metabolism ; therapy ; Oxidation-Reduction ; Oxidative Stress ; Pregnancy ; Pregnancy Rate ; Proteins ; metabolism

OBJECTIVETo investigate the association of anti-Mullerian hormone (AMH) levels in the follicular fluid and serum with the outcomes of in vitro fertilization-embryo transfer (IVF-ET) cycles in patients with polycystic ovarian syndrome (PCOS).

METHODSSerum and follicular fluid samples were obtained from 30 patients with PCOS and 34 healthy women (control) undergoing IVF/ICSI-ET in our center between October, 2007 and January, 2008. All the subjects received treatment with long luteal-phase down-regulation and controlled ovarian hyperstimulation protocol in IVF cycles, and their clinical characteristics were analyzed. The AMH levels in the serum and follicles fluid samples collected on the day of oocyte retrieval were assayed using an enzyme-linked immunosorbent assay (ELISA) kit.

RESULTSThe two groups showed no significant differences in the mean age, baseline levels of sex hormones, rate of high-quality embryos, implantation rate, pregnancy rate, abortion rate or ongoing pregnancy rate (P>0.05). Despite a significantly lower total gonadotropin dose, PCOS group had a significantly greater number of antral follicles than the control group (P<0.05). The recovery rates of oocytes in PCOS group were significantly lower than that in the control group (P<0.05). AMH levels in the serum and follicle fluid was significantly higher in PCOS group than in the control group (P<0.05), and in both groups, AMH levels in the follicular fluid were significantly higher in pregnant women than in non-pregnant women (P<0.05). AMH level in the follicular fluid was significantly correlated with the implantation rate in both PCOS and control groups (P<0.05).

CONCLUSIONAMH level in the serum and follicle fluid on the day of oocyte retrieval is predictive of the treatment outcome of controlled ovarian hyperstimulation in POCS patients but not of pregnancy outcomes after IVF-ET.

Adult ; Anti-Mullerian Hormone ; blood ; metabolism ; Case-Control Studies ; Female ; Fertilization in Vitro ; methods ; Follicular Fluid ; metabolism ; Humans ; Polycystic Ovary Syndrome ; blood ; metabolism ; therapy ; Pregnancy ; Pregnancy Outcome

OBJECTIVETo study the differentially expressed proteins between mature and antral follicles and identify the proteins crucial for follicle development and oocyte fertilization.

METHODSMature follicular fluids were obtained from 48 women after oocyte collection during in vitro fertilization (IVF), and antral follicular fluids were collected from 21 women by follicular puncture. The proteins in the follicular fluids were detected by surface-enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS) and weak cation-exchange protein chip (WCX-2). The data were read with PBSII-C type protein chip reader and analyzed with Biomarker Wizard and Biomarker Patterns Software of Ciphergen Company.

RESULTSIn comparison with those in the antral follicular fluid, the concentration of 4 proteins were significantly different in mature follicular fluid, including two up-regulated and two down-regulated proteins.

CONCLUSIONSignificant variation occurs in the proteomics of mature follicular fluid, and the differentially expressed proteins may have close relation to follicle development and oocyte fertilization.

Adult ; Female ; Fertilization in Vitro ; Follicular Fluid ; metabolism ; Humans ; Ovarian Follicle ; metabolism ; Protein Array Analysis ; Proteome ; metabolism ; Proteomics ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Young Adult

OBJECTIVETo study the effect of Chinese drugs for supplementing qi-blood and nourishing Gan-Shen (CD) on transforming growth factor beta1 (TGF beta1) and sex hormone levels in the follicle fluid of women during in vitro fertilization-embryo transfer (IVF-ET) cycle.

METHODSEighty-six women undergoing IVF-ET were randomly assigned to two groups, 45 in the CD group and 41 in the control group. All received the standard regimen for promoting ovulation, but to women in the CD group, 1-week treatment of Cuhuangti Granule was administered during the period of gonadotropin releasing hormone agonist (GnRH-a) down-regulation, and Jinghou Zengzhi Granule was given from time of ovulation promoting with Gn to the day of HCG administration. On the day of oocyte retrieval, TGFbeta1 was detected using enzyme linked immunosorbent assay (ELISA), estradiol (E2), progestone (P), and luteinizing hormone (LH) detected by radioimmunoassay.

RESULTSFollicle fluid contents of TGFbeta1 and LH in the CM group (3.25 +/- 1.11 pg/L and 0.89 +/- 0.45 IU/L) were obviously higher than those in the control group (2.21 +/- 1.08 pg/L and 0.57 +/- 0.42 IU/L, both P < 0.05).

CONCLUSIONChinese drugs for supplementing qi-blood and nourishing Gan-Shen could significantly improve TGFbeta1 and LH levels in the follicle fluid of women, thus enhancing the embryo implantation rate.

Adult ; Drugs, Chinese Herbal ; pharmacology ; Embryo Transfer ; Female ; Fertilization in Vitro ; Follicular Fluid ; metabolism ; Humans ; Luteinizing Hormone ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Young Adult

PURPOSE: To investigate the direct relationship between the follicular fluid (FF) level of soluble human leukocyte antigen G (HLA-G) and fertilizability of the corresponding oocyte as well as the morphological quality of the corresponding embryo. MATERIALS AND METHODS: Sixty-three patients were stimulated with recombinant FSH combined with gonadotropin-releasing hormone (GnRH) agonist long (n=5) or antagonist protocol (n=58) for standard in vitro fertilization (IVF). At the time oocyte retrieval, follicular fluid was obtained from single dominant follicle in 63 patients, and the level of soluble HLA-G was measured by sandwich enzyme-liked immunosorbent assay (ELISA). Normal fertilization and individual embryo quality were evaluated, and were graded to four categories by morphological criteria (the embryo with symmetrical blastomeres and no fragmentation were assigned as grade A). Good-quality embryo was defined as those with grade A or B. RESULTS: Soluble HLA-G was not detected in 15 FF samples. In the group with positive FF soluble HLA-G (sHLA-G) (n=48), high levels of sHLA-G (>117.758 U/mL) could predict the failure of fertilization with statistical significance {area under the curve (AUC) 0.676, 95% confidence interval (CI) 0.525-0.804}. However, the FF sHLA-G level was not related with the formation of good-quality embryo. CONCLUSION: High level of FF sHLA-G could predict the fertilization failure of the corresponding oocyte, but was not related with the formation of good-quality embryo.
Adult ; Enzyme-Linked Immunosorbent Assay ; Female ; Follicular Fluid/*metabolism ; HLA-G Antigens/*metabolism ; Humans ; Oocytes/*cytology/physiology ; Ovarian Follicle/*cytology/physiology

OBJECTIVETo explore the mechanism of action of Er'zhi Tiangui Granule (ETG) in improving quality of oocytes by observing metabonomics and level of calcium ion in follicle fluid, and to investigate the impacts of calcium ion, cholinesterase (ChE) and creatinine (CCr) levels in human follicle fluid on the quality of oocytes and outcome of pregnancy in patients after in vitro fertilization and embryo transfer (IVF-ET).

METHODSFifty-seven patients after IVF-ET were randomly assigned to two groups: the trial group (27 patients) and the control group (30 patients), both were treated with conventional Western medical treatment, but ETG and Liuwei Dihuang Granule (LDG) was given respectively to the two groups additionally. Changes of Shen-asthenia syndrome, amount of oocyte obtained, fertilization rate, cleavage rate, high-quality embryo rate and pregnancy rate, levels of calcium ion, ChE and Cr in follicle fluid, and metabonomics in the two groups were observed and compared.

RESULTSAfter treatment, scores of Shen-asthenia syndrome 8.30 +/- 1.46, fertilization rate 0.82 +/- 0.09, cleavage rate 0.97 +/- 0.07 and high-quality embryos rate 0.51 +/- 0.18 in the trial group were all better than those in the control group (9.16 +/- 1.15, 0.74 +/- 0.18, 0.91 +/- 0.10, 0.41 +/- 0.09, respectively, P<0.05); metabonomics principal component analysis showed that in the trial group, the principal component in follicle fluid distributed mainly in the section I, and that in the control group distributed mainly in the section II, showing significant difference between the two groups; while in most pregnant patients, it distributed in the section I. The Ca2+ concentration in the trial group was significantly higher than that in the control group (P<0.05). Levels of ChE and Cr in the trial group were higher than those in the control group, but the difference between them was insignificant.

CONCLUSIONETG is better than LDG in regulating metabonomics, Ca2+ concentration, improving the quality of oocyte and embryo, and increasing pregnant rate in patients after IVF-ET.

Adult ; Calcium ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Embryo Transfer ; Female ; Fertilization in Vitro ; Follicular Fluid ; drug effects ; metabolism ; Humans ; Infertility, Female ; etiology ; metabolism ; Metabolomics ; Oocytes ; metabolism ; Phytotherapy ; Pregnancy