A transcriptome approach evaluating the effects of Atractylenolide Ⅰ on the secretion of estradiol and progesterone in feline ovarian granulosa cell

Abstract Atractylodes macrocephala Koidz (AMK) as eartraditional oriental medicine has been used in the treatment of threatened abortion. Atractylenolide I (AT-I) is one of the major bioactive components of AMK. This study aimed to investigate the effect of AT-I on the secretion of estradiol (E 2 ) and progesterone (P 4 ) of feline ovarian granulosa cells (FOGCs) which is necessary for pregnancy. At first, the prolifeation of FOGCs after AT-I treatment was measured by CCK-8. Then, the synthesis of E 2 and P 4 were measured by ELISA. Lastly, transcriptome sequencing was used to detect the DEGs in the FOGCs, and RNA-Seq results were verified by RT-qPCR and biochemical verification. It was found that AT-I could promote proliferation and the secretion of E 2 and P 4 in FOGCs; after AT-I treatment, 137 significantly DEGs were observed, out of which 49 were up-regulated and 88 down-regulated. The DEGs revealed significant enrichment of 52 GO terms throughout the differentiation process ( P < 0.05) as deciphered by Gene Ontology enrichment analysis. Kyoto Encyclopedia of Genes and Genomes analysis manifested that the DEGs were successfully annotated as members of 155 pathways, with 23 significantly enriched ( P < 0.05). A relatively high number of genes were enriched for the cholesterol metabolism, ovarian steroidogenesis, and biosynthesis of unsaturated fatty acids. Furthermore, the contents of the total cholesterol and low-density lipoprotein cholesterol were decreased by AT-I treatment in the cell culture supernatant. The results indicated that AT-I could increase the ability of FOGCs to secrete E 2 and P 4 , which might be achieved by activation of cholesterol metabolism.