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

Abstract

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 (E2) and progesterone (P4) 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 E2 and P4 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 E2 and P4 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 E2 and P4, which might be achieved by activation of cholesterol metabolism.