黄苑,
苏晓鸥,
张维,
齐香荣,
林刚
中国农业科学院农业质量标准与检测技术研究所, 农业部农产品质量安全研究重点实验室, 北京 100081

作者简介: 黄苑(1993-),女,在读研究生,研究方向为农产品质量与食物安全,E-mail:13913004127@163.com.

基金项目: 国家重点研发计划(2017YFC1600300)


中图分类号: X171.5

Effects of PCB118 on GnRH Secretion and Proteins Expression of Kisspeptin/GPR54 Signaling Pathway in GT1-7 Cells

Huang Yuan,
Su Xiao-Ou,
Zhang Wei,
Qi Xiangrong,
Lin Gang
Institute of Quality Standard and Testing Technology for Agro-products of Chinese Academy of Agricultural Sciences, Key Laboratory of Agricultural Product Quality and Safety Research of Ministry of Agriculture, Beijing 100081, China

CLC number: X171.5

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摘要:为研究多氯联苯118(polychlorinated biphenyl 118, PCB118)对GT1-7细胞的毒性作用，探讨其对促性腺激素释放激素(gonadotropin-releasing hormone, GnRH)分泌的影响及相关机制，在PCB118作用GT1-7细胞后，分析了细胞存活率、GnRH分泌水平以及Kisspeptin/GPR54信号通路中关键蛋白的表达水平。结果表明，PCB118(0.05~50 000 nmol·L^-1)分别作用GT1-7细胞6、12、24和48 h后，细胞存活率随PCB118浓度的增加和作用时间的延长而显著降低。但只有在高浓度(50 000 nmol·L^-1)条件下，PCB118才能显著促进GT1-7细胞释放乳酸脱氢酶(lactate dehydrogenase, LDH)。PCB118作用GT1-7细胞6 h后，5、500和50 000 nmol·L^-1组能显著抑制GnRH的分泌，并降低Kisspeptin/GPR54信号通路中GnRH、G蛋白偶联受体54(G-protein-coupled receptor 54, GPR54)、磷脂酶C(phospholipase C, PLC)和蛋白激酶C(protein kinase C, PKC)等关键蛋白的表达水平；作用24 h，500和50 000 nmol·L^-1 PCB118能显著抑制GnRH的分泌，并降低GnRH、GPR54、PLC和PKC等关键蛋白的表达水平。研究表明：PCB118对GT1-7细胞具有细胞增殖毒性作用，呈现一定的剂量和时间效应；PCB118能在一定程度上抑制GT1-7细胞GnRH的合成与分泌，并下调Kisspeptin/GPR54信号通路中GnRH、GPR54、PLC和PKC等关键蛋白的表达水平，推测Kisspeptin/GPR54信号通路可能参与介导了PCB118对GT1-7细胞GnRH分泌的抑制作用。
关键词: PCB118/
GT1-7细胞/
Kisspeptin/GPR54/
GnRH

Abstract:To explore the underlying mechanism of dioxin-like polychlorinated biphenyl 118 (PCB118) on the secretion of gonadotropin-releasing hormone (GnRH), GT1-7 cells were exposed to different concentrations of PCB118. Cell viability, release of GnRH and expression of associated proteins in Kisspeptin/GPR54 signaling pathway were investigated. The results showed that PCB118 (0.05-50 000 nmol·L^-1) significantly reduced cell viability of GT1-7 cells in dose- and time-dependent manners after exposure for 6 h, 12 h, 24 h and 48 h. Interestingly, decreased release of lactate dehydrogenase (LDH) was only observed under the treatment of 50 000 nmol·L^-1 PCB118. Exposure to 5, 500 and 50 000 nmol·L^-1 PCB118 for 6 h significantly decreased the levels of GnRH in the culture media with significant down expression of proteins in Kisspeptin/GPR54 signaling pathway, including GnRH, GPR54, PLC and PKC in GT1-7 cells. Similarly, significantly-decreased GnRH levels and down regulation of such proteins were observed after exposure to 500 or 50 000 nmol·L^-1 PCB118 for 24 h. Taken together, PCB118 shows cytotoxic effects on GT1-7 cells in dose- and time-dependent manners. These results indicate that PCB118 might affect the secretion of GnRH through inhibiting the expression of associated proteins in Kisspeptin/GPR54 signaling pathway.
Key words:PCB118/
GT1-7 cells/
Kisspeptin/GPR54/
GnRH.