RNA-seq based gene expression analysis of ovarian granulosa cells exposed to zearalenone in vitro: significance to steroidogenesis
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Guo-Liang Zhang1,2,3, Rui-Qian Zhang1, Xiao-Feng Sun1, Shun-Feng Cheng1, Yu-Feng Wang1, Chuan-Liang Ji2, Yan-Zhong Feng4, Jie Yu2, Wei Ge1, Yong Zhao1, Shi-Duo Sun3, Wei Shen1 and Lan Li1
1College of Animal Science and Technology, Qingdao Agricultural University, Qingdao Shandong 266109, China
2National Engineering Research Center for Gelatin-based Traditional Chinese Medicine, Dong-E-E-Jiao Co., Ltd, Liaocheng Shandong 252000, China
3College of Animal Science and Technology, Northwest A&F University, Yangling Shaanxi 712100, China
4Institute of Animal Sciences, Heilongjiang Academy of Agricultural Sciences, Harbin Heilongjiang 150086, China
Lan Li, email: [email protected]
Keywords: porcine, granulosa cells, steroidogenesis, gene expression, RNA-seq
Received: April 16, 2017 Accepted: June 20, 2017 Published: July 31, 2017
Zearalenone (ZEA) is a natural contaminant of various food and feed products representing a significant problem worldwide. Since the occurrence of ZEA in grains and feeds is frequent, the present study was carried out to evaluate the possible effects of ZEA on steroid production and gene expression of porcine granulosa cells, using RNA-seq analysis. Porcine granulosa cells were administered 10 μM and 30 μM ZEA during 72 h of culture in vitro. Following ZEA treatment the gene expression profile of control and exposed granulosa cells was compared using RNA-seq analysis. The results showed that in the exposed granulosa cells ZEA significantly altered the transcript levels, particularly steroidogenesis associated genes. Compared with the control group, 10 μM and 30 μM ZEA treatment significantly increased the mRNA expression of EDN1, IER3, TGFβ and BDNF genes and significantly reduced the mRNA expression of IGF-1 and SFRP2 genes. In particular, ZEA significantly decreased the expression of genes essential for estrogen synthesis including FSHR, CYP19A1 and HSD17β in granulosa cells. Furthermore, Q-PCR and Western-blot analysis also confirmed reduced expression of these genes in ZEA exposed granulosa cells. These effects were associated with a significant reduction of 17β-estradiol concentrations in the culture medium of granulosa cells. Collectively, these results demonstrated a concretely deleterious effect of ZEA exposure on the mRNA expression of steroidogenesis related genes and the production of steroid hormones in porcine ovarian granulosa cells in vitro.
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