Subsequent to the administration of 0.1 mg GnRH antagonist (GnRH-a, Tryptorelin, Ferring, Germany) on day 21 of menstruation, it was used throughout the COH period. rFSH (Gonal F, Serono. Ltd., Switzerland) was given when the patients reached the downregulation criteria. The downregulation criteria were as follows: FSH <5 mIU/mL, LH <5 mIU/mL, E2 <30 pg/mL, and P <0.6 ng/mL; endometrial ≤5 mm; and antral follicle diameter 4–7 mm. Gn was used to induce follicle development. The administration of a starting dose of 225–300 IU of human recombinant FSH (rFSH, Gonal F, Serono. Ltd, Switzerland) lead to a subsequent adjustment of gonadotropin (Gn) use according to follicular growth. The starting dose is according to age, AFC, and basal hormone levels.
Gnrh a
Manufactured by Ferring
Sourced in Germany
GnRH-a is a laboratory equipment product used for research purposes. It is a synthetic analogue of the naturally occurring gonadotropin-releasing hormone (GnRH). GnRH-a is primarily used in research to study the regulation of the hypothalamic-pituitary-gonadal axis.
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4 protocols using gnrh a
1
GnRH Antagonist Protocol for Controlled Ovarian Hyperstimulation
2
GnRH Antagonist Ovarian Stimulation
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The patients were injected with 3.75 mg GnRH antagonist (GnRH-a, Tryptorelin, Ferring, Germany) on the 2nd day of menstruation if the ultrasound did not find cysts and follicles >10 mm. The patients will visit the hospital 28 days after the injection to be examined by ultrasound and to check the serum FSH, LH, E2, and progesterone (P) levels. A starting dose of 225–300 IU of human recombinant FSH (rFSH, Gonal F, Serono. Ltd., Switzerland) was administered, with subsequent adjustment of gonadotropin (Gn) use according to follicular growth. The starting dose is according to age, AFC, and basal hormone levels.
3
GnRHa-Assisted Controlled Ovarian Hyperstimulation
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In PCOS patients, GnRHa (0.1 mg/day; Ferring, Kiel, Germany) was administered on the 21st day of spontaneous menstruation or a progestin-induced withdrawal bleed until hCG day. COH began on menstrual cycle day 2 after GnRHa administration with rFSH (Serono, Geneva, Switzerland). The starting dose of rFSH was 150 IU/day for all patients in both groups and this dose was subsequently adjusted depending on the ovarian response, as assessed by E2 levels combined with ultrasound. As soon as at least two leading follicles reached a mean diameter of ≥18 mm, 10000 IU of hCG (Pregnyl, Organon, The Netherlands) was administered and oocyte retrieval was performed by vaginal ultrasound 34–36 h after hCG administration.
4
Ovarian Stimulation in Sprague-Dawley Rats
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Speci c pathogen-free female Sprague-Dawley (SD) rats aged 7 weeks were purchased from the Department of Laboratory Animals. They were housed in a clean barrier environment (temperature 24 ± 2°C; relative humidity 50% ± 5%; lights on for 12 h: from 6:00 am to 18:00; with free access to food and water; 3-4 rats per cage). After ve days of adaptation, vaginal smears were observed to determine the estrous cycle and e ciency of COS treatment (Fig. 1A). Rats that showed incomplete estrous cycles, tootoo-deeptuitary suppression, hyperstimulation, and failure on mating were excluded.
GnRH-a/HP-hMG/HCG (Group M) and GnRH-a/u-FSH/HCG (Group F) were used to generate OS rats. Gonadotropin-releasing hormone agonist (GnRH-a, Ferring, Denmark) was administered i.p. 4 µg/250-300 g every day at 9 am since post-estrus. High-purity hMG (HP-hMG, MENOPUR, Ferring) or u-FSH (Livzon, China) was administered i.p. 40 IU/200 g at 9 am on day 7 of downregulation. HCG was administered i.p. 1050 IU/200 g 48 h after gonadotropin injection. The rats in the natural cycle (Group NC) were administered the same volume of 4°C saline at the same time. Male and female rats were caged 1:2 on the day of HCG and marked Pd1if vaginal plug or sperm (Fig. 1B) were observed on the next day morning. We collected ovaries, uterus, and UF at Pd5.
GnRH-a/HP-hMG/HCG (Group M) and GnRH-a/u-FSH/HCG (Group F) were used to generate OS rats. Gonadotropin-releasing hormone agonist (GnRH-a, Ferring, Denmark) was administered i.p. 4 µg/250-300 g every day at 9 am since post-estrus. High-purity hMG (HP-hMG, MENOPUR, Ferring) or u-FSH (Livzon, China) was administered i.p. 40 IU/200 g at 9 am on day 7 of downregulation. HCG was administered i.p. 1050 IU/200 g 48 h after gonadotropin injection. The rats in the natural cycle (Group NC) were administered the same volume of 4°C saline at the same time. Male and female rats were caged 1:2 on the day of HCG and marked Pd1if vaginal plug or sperm (Fig. 1B) were observed on the next day morning. We collected ovaries, uterus, and UF at Pd5.
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