Suppression of miR-1197e3p attenuates H 2 O 2 -induced apoptosis of goat luteinized granulosa cells via targeting PPARGC1A Guo-Min Zhang a, b , Shi-Yu An b , M.A. El-Samahy b , Yan-Li Zhang b , Yong-Jie Wan b , Zi-Yu Wang b , Shen-Hua Xiao b , Fan-Xing Meng b , Feng Wang b, * , Zhi-Hai Lei a, ** a Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China b Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, 210095, China article info Article history: Received 17 December 2018 Received in revised form 6 April 2019 Accepted 6 April 2019 Available online 11 April 2019 Keywords: Goat luteinized granulosa cells miR-1197e3p PPARGC1A Mitochondrial-dependent apoptosis pathway abstract Peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PPARGC1A) acts as a powerful coactivator of many transcriptional factors that relate to granulosa cell (GC) apoptosis. In this study, the miRNAs mediating goat follicular atresia and luteinized granulosa cell (LGC) apoptosis induced by hydrogen peroxide (H 2 O 2 ) via PPARGC1A were investigated. Our results showed that miR-1197e3p tar- geted PPARGC1A was predicted by bioinformatics algorithm and verified by luciferase reporter assay. In addition, miR-1197e3p promoted goat LGC apoptosis via PPARGC1A through mitochondrial-dependent apoptosis pathway, and these effects could be restored by PPARGC1A overexpression. Moreover, H 2 O 2 - induced LGC apoptosis significantly upregulated miR-1197e3p expression and downregulated PPARGC1A level. Pretreatment of miR-1197e3p inhibitor alleviated LGC apoptosis induced by 400 mMH 2 O 2 for 12 h, and preserved the mitochondrial membrane potential by increasing PPARGC1A expression. In conclusion, miR-1197e3p might act as an essential regulator of goat LGC apoptosis potentially via the mitochondrial- dependent apoptosis pathway by targeting PPARGC1A. © 2019 Elsevier Inc. All rights reserved. 1. Introduction In mammals, less than 1% of the ovarian follicles will eventually ovulate, and most of the follicles will undergo atresia during follicular growth and development [1]. Follicular atresia limits the number of ovulations, which restricts the full reproductive poten- tial, thus the follicular atresia has been a hotspot of animal repro- duction research [2]. Follicular atresia is mainly attributed to granulosa cell (GC) apoptosis, which was regulated by reproductive hormones, cytokines and apoptotic related factors [3]. However, the internal molecular and cellular mechanisms responsible for the activation and regulation of GC apoptosis still need further investigations. Small noncoding microRNAs (miRNAs) are known to negatively regulate gene translation by binding complementary mRNA targets in the three prime untranslated regions (3 0 -UTR) [4], and have been identified and characterized in many biological and cellular pro- cesses [5]. Using extensive microarray profiling of ovarian tissues or cells, many studies have demonstrated distinct miRNA expression profiles along with follicular development, including primordial formation, follicular recruitment and selection, follicular atresia, and luteinization [6]. In addition, several functional studies have identified miRNAs could regulate GC apoptosis by directly targeting genes involved in apoptosis signaling pathways [6,7], while the mechanisms of miRNA regulating GC apoptosis and follicular atresia have remained to be explored. Importantly, despite of extensive researches, studies focused on the roles of miRNAs in the regulation of goat ovarian development, especially in GC apoptosis and follicular atresia are still limited. Peroxisome proliferator-activated receptor gamma coactivator- 1 alpha (PPARGC1A) is a transcription coactivator that interacts with multiple transcription factors involved in energy metabolism and mitochondrial biogenesis. A number of studies have revealed that the aberrant expression of PPARGC1A impaired mitochondrial biogenesis, which was vital for cell survival [8,9]. Mitochondrial * Corresponding author. Jiangsu Livestock Embryo Engineering Laboratory, Col- lege of Animal Science and Technology, Nanjing Agricultural University, No.1 Weigang, Nanjing, China. ** Corresponding author. College of veterinary medicine, Nanjing Agricultural University, No.1 Weigang, Nanjing, China. E-mail addresses: caeet@njau.edu.cn (F. Wang), leizh@njau.edu.cn (Z.-H. Lei). Contents lists available at ScienceDirect Theriogenology journal homepage: www.theriojournal.com https://doi.org/10.1016/j.theriogenology.2019.04.008 0093-691X/© 2019 Elsevier Inc. All rights reserved. Theriogenology 132 (2019) 72e82