Department of Endocrinology, XuZhou Children's Hospital, Xuzhou, Jiangsu, China (mainland).
Med Sci Monit. 2020 May 26;26:e922860. doi: 10.12659/MSM.922860.
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 ERß, CD36, and GPR54 in GT1-7 cells, and the expression of ERß 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 ß (ERß), fatty acid translocase protein (CD36), and G Protein-Coupled Receptors 54 (GPR54) in GT1-7 cells. After estrogen receptors ß (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.
本研究旨在探讨高糖高脂环境对 GT1-7 细胞中雌激素受体和性早熟相关基因的影响。
本研究采用 CCK8 法检测氨基葡萄糖和棕榈酸处理后 GT1-7 细胞的活力,TUNEL 法检测细胞凋亡水平。采用 ELISA 和 RT-qPCR 检测 GnRH 的表达水平。采用 RT-qPCR 和 Western blot 检测 GT1-7 细胞中 ERß、CD36 和 GPR54 的表达,采用免疫组化分析检测 ERß 的表达。最后,在 GT1-7 细胞中加入干预药物他莫昔芬后,采用 ELISA 检测 GnRH 的表达水平,Western blot 分析检测 GPR54 和 GnRH 的表达。
高糖高脂组的 GnRH 分泌随时间持续增加,并在 18 小时达到峰值,而 GnRH 基因表达在 12 小时达到峰值。高糖高脂条件还显著增加了 GT1-7 细胞中雌激素受体β(ERß)、脂肪酸转运蛋白(CD36)和 G 蛋白偶联受体 54(GPR54)的水平。抑制雌激素受体β(ER)后,12 小时和 18 小时 GnRH 分泌和 GPR54 表达均降低。
本研究表明,高糖高脂环境促进了 GT1-7 细胞中 GnRH 的分泌和 ER 的表达,以及与性早熟相关基因的表达。