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The effects of phospholipase C on oestradiol and progesterone secretion in porcine granulosa cells cultured in vitro
Author(s) -
Chen Huali,
Yang Youfu,
Wang Youlin,
He Yamei,
Duan Jiaxin,
Cheng Jianyong,
Li Qingwang
Publication year - 2019
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/rda.13517
Subject(s) - medicine , endocrinology , phospholipase c , activator (genetics) , granulosa cell , in vitro , secretion , phospholipase , biology , phospholipase a2 , chemistry , phospholipase d , signal transduction , receptor , ovary , enzyme , microbiology and biotechnology , biochemistry
Abstract Granulosa cells play important roles in the regulation of ovarian functions. Phospholipase C is crucial in several signalling pathways and could participate in the molecular mechanisms of cell proliferation, differentiation and ageing. The objective of this study was to identify the effects of phospholipase C on the steroidogenesis of oestradiol and progesterone in porcine granulosa cells cultured in vitro. Inhibitor U73122 or activator m‐3M3FBS of phospholipase C was added to the in vitro medium of porcine granulosa cells, respectively. The secretion of oestradiol decreased after 2 hr, 8 hr, 12 hr, 24 hr and 48 hr of treatment with 500 nM U73122 ( p < .05) and decreased after 2 hr of treatment in the 500 nM m‐3M3FBS addition group ( p < .05). The secretion of progesterone increased after 4 hr of treatment with 500 nM U73122 ( p < .05) and increased after 2 hr and 8 hr of treatment in the 500 nM m‐3M3FBS addition group ( p < .05). The ratio of oestradiol to progesterone decreased at each time point, except 8 hr after the addition of 500 nM U73122 ( p < .05). The ratio of oestradiol to progesterone decreased after 2 hr ( p < .05) of treatment with 500 nM m‐3M3FBS. In genes that regulate the synthesis of oestradiol or progesterone, the mRNA expression of CYP11A1 was markedly increased ( p < .05), and the mRNA expression of other genes did not change significantly in the U73122 treatment group, while the addition of m‐3M3FBS did not change those genes significantly despite the contrary trend. Our results demonstrated that phospholipase C can be a potential target to stimulate the secretion of oestradiol and suppress progesterone secretion in porcine granulosa cells cultured in vitro, which shed light on a novel biological function of phospholipase C in porcine granulosa cells.