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Войти в исследования European journal of obstetrics, gynecology, and reproductive biology · 2021
Influence of pretreatment of insulin on the phosphorylation of extracellular receptor kinase by gonadotropin-releasing hormone and gonadotropins in cultured human granulosa cells.
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Ko Yuri, Kim Ju Hee, Lee Sa Ra, Kim Sung Hoon, Chae Hee Dong
Аннотация
OBJECTIVE: To investigate the influence of pretreatment of insulin on the phosphorylation of ERK1/2 by gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) in cultured human granulosa cells.
STUDY DESIGN: Human granulosa cells were collected from 20 women (age: 20-35 years) who underwent controlled ovarian hyperstimulation for in vitro fertilization and embryo transfer at Asan Medical Center (Seoul, South Korea). The presence of the receptors for insulin, GnRH, FSH, and LH in human granulosa cells was identified by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). The granulosa cells were treated with 10 nM insulin or 10 nM GnRH for 5 min or 30 min and with 10 nM FSH or 10 nM LH for 24 h or 48 h. The cells were also pretreated with insulin for 30 min prior to treatment with GnRH, FSH, or LH. Western blot analysis was used to analyze ERK1/2 phosphorylation.
RESULTS: RT-PCR showed that the receptors for insulin, GnRH, FSH, and LH were expressed in human granulosa cells. Insulin, GnRH, FSH, and LH could activate ERK1/2 phosphorylation. Pretreatment with insulin inhibited ERK1/2 phosphorylation induced by GnRH and FSH while augmenting ERK1/2 phosphorylation induced by LH.
CONCLUSIONS: Insulin might have a negative effect on GnRH and FSH regulation by attenuating the action of GnRH and FSH in the phosphorylation of ERK1/2 in human granulosa cells. In contrast, insulin might have a positive effect on LH regulation by potentiating the action of LH in the phosphorylation of ERK1/2. Our results showed that insulin is clearly an important regulator of human reproductive function at the ovarian level.
STUDY DESIGN: Human granulosa cells were collected from 20 women (age: 20-35 years) who underwent controlled ovarian hyperstimulation for in vitro fertilization and embryo transfer at Asan Medical Center (Seoul, South Korea). The presence of the receptors for insulin, GnRH, FSH, and LH in human granulosa cells was identified by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). The granulosa cells were treated with 10 nM insulin or 10 nM GnRH for 5 min or 30 min and with 10 nM FSH or 10 nM LH for 24 h or 48 h. The cells were also pretreated with insulin for 30 min prior to treatment with GnRH, FSH, or LH. Western blot analysis was used to analyze ERK1/2 phosphorylation.
RESULTS: RT-PCR showed that the receptors for insulin, GnRH, FSH, and LH were expressed in human granulosa cells. Insulin, GnRH, FSH, and LH could activate ERK1/2 phosphorylation. Pretreatment with insulin inhibited ERK1/2 phosphorylation induced by GnRH and FSH while augmenting ERK1/2 phosphorylation induced by LH.
CONCLUSIONS: Insulin might have a negative effect on GnRH and FSH regulation by attenuating the action of GnRH and FSH in the phosphorylation of ERK1/2 in human granulosa cells. In contrast, insulin might have a positive effect on LH regulation by potentiating the action of LH in the phosphorylation of ERK1/2. Our results showed that insulin is clearly an important regulator of human reproductive function at the ovarian level.