LEAD ACCUMULATION IN HUMAN OVARIAN FOLLICULAR FLUID, AND IN VITRO EFFECT OF LEAD ON PROGESTERONE PRODUCTION BY CULTURED HUMAN OVARIAN GRANULOSA CELLS

Abstract Purpose: Polycystic ovary syndrome (PCOS) is considered as one of the most common endocrine disorder with heterogeneity. There are also reports that liver receptor homolog 1 [LRH-1 or nuclear receptor subfamily 5 group A member 2 (NR5A2)] plays an important role in the reproductive system. But up to now, there are no reports related to the link with PCOS and LRH-1. In this study, we aimed to detect the LRH-1 expression in the ovarian granulosa cell of PCOS patients and explore the potential relationship between LRH-1 and PCOS. Methods: 146 follicular fluid sample were collected in this study, including 72 from PCOS patients and 74 from control patients who underwent intracytoplasmic sperm injection (ICSI) or in vitro fertilization-embryo transfer (IVF-ET). The ovarian granulosa cells were extracted from the patient's follicular fluid by magnetic-activated cell sorting (MACS) method, and the real-time quantitative PCR (qRT-PCR) was used to measure the expression of LRH-1 in ovarian granulosa cells. Then we analyzed the correlation between the expression level of LRH-1 and the clinical characteristics of patient by using Pearson Correlation analysis. Results: The expression of LRH-1 was significantly higher in PCOS patients ovarian granulosa cells than that in the control patients [vs(1.38±0.47)vs(1.03±0.32), t=5.327, p<0.0001], and it was positively correlated with antral follicles counting (AFC) (r=0.3607, p<0.0001)and the serum AMH(r=0.2662, p=0.0012)\LH(r=0.2518, p=0.0022)\T(r=0.2516, p=0.0022) in all patients. No statistical significance between LRH-1 and BMI, FSH, HOMA-IR, DHE-S, progesterone. Conclusions: Compared with the control group, we found that LRH-1 was highly expressed in the ovarian granulosa cells of PCOS patients. Our study has revealed the relationship between the LRH-1 expression and PCOS, which suggested that LRH-1 may play an important role in ovulation disorders. While this finding provided new ideas for the study of the pathogenesis, it also provided a theoretical basis for the clinical diagnosis and treatment for PCOS.

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Fibronectin production by chicken granulosa cells in vitro: effect of follicular development

Acta Endocrinologica ◽

10.1530/acta.0.1270466 ◽

1992 ◽

Vol 127 (5) ◽

pp. 466-470 ◽

Cited By ~ 11

Author(s):

Elikplimi K Asem ◽

Jacqueline A Carnegie ◽

Benjamin K Tsang

Keyword(s):

Granulosa Cells ◽

Follicular Development ◽

Enzyme Linked Immunosorbent Assay ◽

Dependent Manner ◽

Ovarian Granulosa Cells ◽

Preovulatory Follicles ◽

Vitro Effect ◽

Dose Dependent

In vitro studies were conducted to investigate the role of chicken ovarian granulosa cells in the production of fibronectin, a component of the basal lamina of ovarian follicles. Collagenase dispersed granulosa cells obtained from the first (F1; about 35 mm in diameter) and third (F3; 15–20 mm in diameter) largest preovulatory follicles, as well as from a pool of small yellow follicles (SF; 6–10 mm in diameter), were incubated in serum-free medium-199 for 24 to 96 h in the absence and presence of luteinizing hormone (LH) or forskolin. Fibronectin secreted in the medium was quantitated by enzyme linked immunosorbent assay. Basal fibronectin production (which increased with the duration of incubation) was significantly greater (p<0.001) in granulosa cells derived from mature follicle (F1) than in F3 or SF cells. Both LH and forskolin stimulated fibronectin production in SF and F3 cells in a dose-dependent manner; however, they were without effect in F1 cells. The magnitude of increase in fibronectin production elicited by LH or forskolin was greater in SF cells than in F3 cells. The cytoplasm of cultured granulosa cells taken at all stages of follicular development stained positively for fibronectin. These findings indicate that chicken granulosa cells produce fibronectin. This ability is acquired early in follicular development and the stimulatory effect of the gonadotropin (LH) diminished as the follicle approached ovulation.

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Effects of Multicycle Gonadotropin-releasing Hormone Antagonist Protocols on Oxidative Stress of Follicular Fluid and Ovarian Granulosa Cells

10.21203/rs.3.rs-136397/v1 ◽

2020 ◽

Author(s):

Yucong Ma ◽

Zhiming Zhao ◽

Guimin Hao ◽

Na Cui ◽

Yanli Fan ◽

...

Keyword(s):

Oxidative Stress ◽

Granulosa Cells ◽

Follicular Fluid ◽

Gonadotropin Releasing Hormone ◽

Clinical Pregnancy ◽

Vitro Fertilization ◽

Releasing Hormone ◽

Ovarian Granulosa Cells ◽

Cycle Group

Abstract Background: Repeated controlled ovarian stimulation (COS) has adverse effects on clinical pregnancy outcomes in in vitro fertilization-embryo transfer (IVF-ET) patients. The effect of repeated antagonist protocols on oxidative stress (OS) in follicular fluid (FF) and ovarian granulosa cells (GC) remains unclear. Objective: To study the effects of repeated multicycle gonadotropin-releasing hormone antagonist (GnRH-ant) protocols on OS markers of FF and ovarian GC. Methods: A total of 145 patients were enrolled and divided into four groups: 1 cycle group (n = 42), 2 cycles group (n = 37), 3 cycles group (n = 45), and 4-5 cycles group (n = 21). The FF and ovarian GC of the patients were collected on the day of last egg retrieval. Levels of 8-hydroxy-2-deoxyguanosine (8-OHdG), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) in FF and ovarian GC were analyzed. Results: With the increase of GnRH-ant protocol cycles, the serum estradiol levels on human chorionic gonadotrophin (hCG) injection day, number of retrieved oocytes, 2PN embryos, and the rates of high-quality embryos, implantation and clinical pregnancy were all significantly decreased, while the gonadotropin (Gn) usage showed an increasing trend. Compared with 1 or 2 cycles, the 8-OHdG and SOD were significantly increased in the 3 to 5 cycles, while the CAT and GSH-Px levels were significantly decreased. Conclusion: Repeated COS with the use of GnRH-ant protocols results in OS and changes the follicle microenvironment of FF and GC, possibly leading to poor IVF outcomes in patients with 3-5 cycles of COS.

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Expression of adrenomedullin by human granulosa lutein cells and its effect on progesterone production

Acta Endocrinologica ◽

10.1530/eje.0.1420671 ◽

2000 ◽

pp. 671-676 ◽

Cited By ~ 20

Author(s):

T Moriyama ◽

T Otani ◽

T Maruo

Keyword(s):

Granulosa Cells ◽

Follicular Fluid ◽

Immunohistochemical Staining ◽

Oocyte Retrieval ◽

Dependent Manner ◽

Vitro Fertilization ◽

Local Factor ◽

Progesterone Production ◽

Midluteal Phase

OBJECTIVE: Adrenomedullin (AM) has diverse functions and is expressed in a variety of tissues. This study was conducted to investigate the expression of AM in the human ovary and its effect on progesterone production by human granulosa lutein cells. DESIGN AND METHODS: Follicular fluid and blood samples were obtained at the time of oocyte retrieval from patients undergoing in vitro-fertilization cycles. Concentrations of AM in follicular fluid and plasma were measured by RIA. Granulosa cells were isolated from follicular fluid and expression of AM mRNA was examined by RT-PCR. Granulosa lutein cells were cultured in vitro and secretion of AM by those cells was determined by immunoprecipitation followed by PAGE. Immunohistochemical staining with human ovaries was carried out, using a specific antibody to AM. Furthermore, the effect of AM on progesterone production by cultured granulosa lutein cells was studied. RESULTS: Concentrations of AM in follicular fluid collected just before ovulation were significantly higher than those in the plasma (P<0.01). AM mRNA was expressed in granulosa cells at the preovulatory stage. Cultured granulosa lutein cells secreted immunoreactive AM. With immunohistochemical staining, it was revealed that AM was most abundantly expressed in granulosa lutein cells at the midluteal phase. No appreciable staining for AM was observed in granulosa cells in primordial and preantral follicles, whereas immunolocalization of AM was noted in granulosa cells of dominant follicles although it was not as prominent as in granulosa lutein cells at the midluteal phase. Furthermore, addition of AM to cultured granulosa lutein cells augmented progesterone secretion in a dose-dependent manner. CONCLUSIONS: These results suggest that AM is transcribed and secreted in human granulosa lutein cells as a local factor to enhance progesterone production by those cells.

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