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

Haiying He; Xiaohui Su; Huiguo Yang; Yingjie Zhang; Chunhui Duan; Ruochen Yang; Fengmei Xie; Yueqin Liu; Wujun Liu

doi:10.5713/ab.23.0448

Animal Bioscience (Oct 2024)

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

Haiying He,
Xiaohui Su,
Huiguo Yang,
Yingjie Zhang,
Chunhui Duan,
Ruochen Yang,
Fengmei Xie,
Yueqin Liu,
Wujun Liu

Affiliations

Haiying He: Department of Animal Science and Biotechnology, Xinjiang Agricultural University, Urumqi, Xinjiang 830052, China
Xiaohui Su: Department of Animal Science and Biotechnology, Xinjiang Agricultural University, Urumqi, Xinjiang 830052, China
Huiguo Yang: Moyu Bibang Sheep Industry Development Co. LTD, Hotan Prefecture, Xinjiang 848100, China
Yingjie Zhang: Department of Animal Science and Biotechnology, Hebei Agricultural University, Baoding, Hebei 071000, China
Chunhui Duan: Department of Animal Science and Biotechnology, Hebei Agricultural University, Baoding, Hebei 071000, China
Ruochen Yang: Department of Animal Science and Biotechnology, Hebei Agricultural University, Baoding, Hebei 071000, China
Fengmei Xie: Department of Animal Science and Biotechnology, Xinjiang Agricultural University, Urumqi, Xinjiang 830052, China
Yueqin Liu: Department of Animal Science and Biotechnology, Hebei Agricultural University, Baoding, Hebei 071000, China
Wujun Liu: Department of Animal Science and Biotechnology, Xinjiang Agricultural University, Urumqi, Xinjiang 830052, China

DOI: https://doi.org/10.5713/ab.23.0448
Journal volume & issue: Vol. 37, no. 10
pp. 1712 – 1725

Abstract

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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<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.

Published in Animal Bioscience

ISSN: 2765-0189 (Print); 2765-0235 (Online)
Publisher: Asian-Australasian Association of Animal Production Societies
Country of publisher: Korea, Republic of
LCC subjects: Science: Zoology
Website: https://www.animbiosci.org/index.php

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