26 May 2020 : Laboratory Research
Effects of High-Glucose and High-Fat Condition on Estrogen Receptor- and Sexual Precocity-Related Genes in GT1-7 Cells
Shuqin Wang1ABCDE, Hong Yao1ABCEF, Lan Ding1BCDEF, Yiqing Gao1BCDE, Pei Wang1BCDE, Ying Xue1ABCDEF*DOI: 10.12659/MSM.922860
Med Sci Monit 2020; 26:e922860
Abstract
BACKGROUND: This study was designed to investigate the effect of high-glucose and high-fat condition on estrogen receptor- and sexual precocity-related genes in GT1-7 cells.
MATERIAL AND METHODS: In this study, CCK8 was used to detect cell viability, and TUNEL assay was used to detect apoptosis levels of GT1-7 cells after treatment with glucosamine and palmitate. The expression level of GnRH was measured by ELISA and RT-qPCR. RT-qPCR and Western blot were used to detect the expression of ERβ, CD36, and GPR54 in GT1-7 cells, and the expression of ERβ was detected using immunohistochemistry analysis. Finally, after adding the intervening drug tamoxifen to GT1-7 cells, the expression level of GnRH was measured by ELISA and Western blot analysis was used to detect the expression of GPR54 and GnRH.
RESULTS: GnRH secretion in the high-fat and high-glucose group increased continuously over time and peaked at 18 h, and GnRH gene expression peaked at 12 h. High-fat and high-glucose conditions also significantly increased the levels of estrogen receptors β (ERβ), fatty acid translocase protein (CD36), and G Protein-Coupled Receptors 54 (GPR54) in GT1-7 cells. After estrogen receptors β (ER) was inhibited, GnRH secretion and GPR54 expression were decreased at 12 h and 18 h.
CONCLUSIONS: Our study demonstrates that high-glucose and high-fat conditions promote the secretion of GnRH and ER and the expression of genes related to sexual precocity in GT1-7 cells.
Keywords: Abdominal Fat, Estrogen Receptor beta, Glucose, CD36 Antigens, Cell Line, Estrogen Receptor alpha, Estrogens, Glucosamine, Gonadotropin-Releasing Hormone, Kisspeptins, Palmitates, Puberty, Precocious, Receptors, Estrogen, Receptors, Kisspeptin-1
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