Effects of prolactin on the proliferation and hormone secretion of ovine granulosa cells

• Published in: Animal bioscience 2024-10, Vol.37 (10), p.1712-1725
• Main Authors: Haiying He, Xiaohui Su, Huiguo Yang, Yingjie Zhang, Chunhui Duan, Ruochen Yang, Fengmei Xie, Yueqin Liu, Wujun Liu
• Format: Article
• Language: English
• Subjects: apoptosis; granulosa cells (gcs); prl receptor (prlr); prolactin (prl); proliferation
• ISSN: 2765-0189; 2765-0235
• DOI: 10.5713/ab.23.0448

Objective The objective of this study was to investigate the effects of prolactin (PRL) on the proliferation and apoptosis of ovine ovarian granulosa cells (GCs) and the secretion of estrogen (E2) and progesterone (P4), as well as to explore the effects of PRL on related genes and proteins. Methods We isolated ovarian GCs from 1-year-old small-tail Han sheep and identified PRL receptor (PRLR) on ovaries and follicle stimulating hormone receptor (FSHR) on ovarian GCs, respectively, using immunohistochemistry. PRL (0, 0.05, 0.50, 5.00 μg/mL) were added to GCs in vitro along with FSH, cell proliferation was measured by cell counting Kit-8 (CCK-8) and apoptosis by flow cytometry. The measurement of E2 and P4 content by enzyme-linked immunosorbent assays after 48 h and 72 h. The expression of functional genes and proteins was identified by real-time quantitative polymerase chain reaction (RT-qPCR) and Western-blot after 48 h. Results PRLR was expressed in both follicular GCs and corpus luteum, whereas FSHR was expressed specifically. The proliferative activity was lower on day 1 while higher on day 4 and day 5. The apoptosis rate of GCs in the 0.05 μg/mL group was significantly higher than that in the control group after treatment with PRL for 24 h (p