Acta Veterinaria et Zootechnica Sinica ›› 2024, Vol. 55 ›› Issue (1): 169-178.doi: 10.11843/j.issn.0366-6964.2024.01.017

• ANIMAL BIOTECHNOLOGY AND REPRODUCTION • Previous Articles     Next Articles

Effect of miR-24-3p on Estradiol Synthesis in Porcine Granulosa Cells

SHI Shengjie1, WANG Liguang1, GAO Lei1, CAI Chuanjiang1, HE Weixian2, CHU Guiyan1*   

  1. 1. College of Animal Science and Technology, Northwest A & F University, Yangling 712100, China;
    2. Qiquan Agriculture and Animal Husbandry Group Co. LTD., Suining 629000, China
  • Received:2023-08-10 Online:2024-01-23 Published:2024-01-24

Abstract: This study was conducted to investigate the effects of miR-24-3p on estradiol synthesis in porcine granulosa cells. In this experiment, ovarian tissue of 180-day old healthy sows (n=20) was collected, and 20 pairs of ovaries were collected for isolation and culture of granulosa cells in each experiment, and mimics and inhibitor of miR-24-3p were transfected into granulosa cells. The ELISA, RT-qPCR, Western blot, and double luciferase reporting tests were used to explore the effect of miR-24-3p on estradiol synthesis in porcine granulosa cells. The results showed that overexpression of miR-24-3p could significantly promote estradiol synthesis (P<0.01), and up-regulate the transcription and translation of StAR, CYP19A1 and CYP11A1 (P<0.05). Interfering with miR-24-3p significantly inhibited the synthesis of estradiol (P<0.05) and significantly down-regulated the mRNA and protein levels of CYP11A1 and CYP19A1 (P<0.05). Further studies showed that TOP1 was the direct target gene of miR-24-3p, overexpression of miR-24-3p could significantly inhibit the expression of TOP1 (P<0.05), and interference with miR-24-3p could significantly up-regulate the expression of TOP1 (P<0.05). What’s more, overexpression of TOP1 could weaken the promoting effect of miR-24-3p on estradiol synthesis in granulosa cells (P<0.05). In summary, miR-24-3p inhibits mRNA and protein levels of TOP1, and promotes the expression level of genes related to estradiol synthesis, thus promoting estradiol synthesis in granulosus cells by targeting TOP1. Since the estradiol synthesis capacity of granulosa cells directly affects the developmental state of ovarian follicles, this study provides a theoretical basis for screening miRNA that can improve reproductive performance of sows.

Key words: miR-24-3p, pigs, granulosa cells, estradiol

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