Animal Bioscience

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
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Effects of prolactin on the proliferation and hormone secretion of ovine granulosa cells in vitro

  • Haiying He (Department of Animal Science and Biotechnology, Xinjiang Agricultural University) ;
  • Xiaohui Su (Department of Animal Science and Biotechnology, Xinjiang Agricultural University) ;
  • Huiguo Yang (Moyu Bibang Sheep Industry Development Co. LTD, Hotan Prefecture) ;
  • Yingjie Zhang (Department of Animal Science and Biotechnology, Hebei Agricultural University) ;
  • Chunhui Duan (Department of Animal Science and Biotechnology, Hebei Agricultural University) ;
  • Ruochen Yang (Department of Animal Science and Biotechnology, Hebei Agricultural University) ;
  • Fengmei Xie (Department of Animal Science and Biotechnology, Xinjiang Agricultural University) ;
  • Yueqin Liu (Department of Animal Science and Biotechnology, Hebei Agricultural University) ;
  • Wujun Liu (Department of Animal Science and Biotechnology, Xinjiang Agricultural University)
  • Received : 2023.10.28
  • Accepted : 2024.04.01
  • Published : 2024.10.01
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Abstract

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 ㎍/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 ㎍/mL group was significantly higher than that in the control group after treatment with PRL for 24 h (p<0.05). Compared with the control group, the secretion of E2 in GCs was reduced significantly (p<0.05) in PRL treatment for 48 h and 72 h, while the secretion of P4 was significantly increased (p<0.05). The mRNA expression levels of PRLR, FSHR, LHR, CYP11A1, HSD3B7, and STAR were significantly higher than those in the control group (p<0.01), and the relative abundance of BCL2 in all PRL group were increased after PRL treatment. Conclusion: PRL promoted the proliferation of GCs and supraphysiological concentrations inhibited apoptosis caused by down-regulation of BAX and up-regulation of BCL2. PRL inhibited E2 by down-regulating CYP19A1 and promoted P4 by up-regulating CYP11A1, STAR, and HSD3B7.

Keywords

Acknowledgement

This work was supported by The China Agriculture Research System under Grant (CARS-38) and (CARS-39-23); The National Key R&D Program of China under Grant (2018YFD0502100); Excellent Youth Program of Hebei Province under Grant (8042018-1081034); Hebei Province Science Foundation for Youths under Grant (C2019204357); and Youth Science and Technology Top Talent Program of Tianshan talents under Grant (2022TSYCJ0023).

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