Phospholipase C inhibits apoptosis of porcine primary granulosa cells cultured in vitro

Abstract

Phospholipase C (PLC) can participate in cell proliferation, differentiation and aging. However, whether it has a function in apoptosis in porcine primary granulosa cells is largely uncertain. The objective of this study was to examine the effects of PLC on apoptosis of porcine primary granulosa cells cultured in vitro. The mRNA expression of BAK, BAX and CASP3, were upregulated in the cells treated with U73122 (the PLC inhibitor). The abundance of BCL2 mRNA, was upregulated, while BAX and CASP3 mRNA expression was decreased after treatment with m-3M3FBS (the PLC activator). Both the early and late apoptosis rate were maximized with 0.5 μM U73122 for 4 h. The rate of early apoptosis was the highest at 4 h and the rate of late apoptosis was the highest at 12 h in the m-3M3FBS group. The protein abundance of PLCβ1, protein kinase C β (PKCβ), calmodulin-dependent protein kinaseII α (CAMKIIα) and calcineurinA (CalnA) were decreased by U73122, and CAMKIIα protein abundance was increased by m-3M3FBS. The mRNA expression of several downstream genes (CDC42, NFATc1, and NFκB) was upregulated by PLC. Our results demonstrated that apoptosis can be inhibited by altering PLC signaling in porcine primary granulosa cells cultured in vitro, and several calcium−sensitive targets and several downstream genes might take part in the processes.

Isorhamnetin Promotes Estrogen Biosynthesis and Proliferation in Porcine Granulosa Cells via the PI3K/Akt Signaling Pathway

“…Apoptosis of GCs leads to follicular atresia . Studies have shown that estrogen (E2) concentration was higher in healthy follicles, while progesterone (P4) and testosterone (T) concentrations were higher in atretic follicles. , Thus, the proliferation, apoptosis, and steroidogenesis of GCs are important for follicular development and atresia.…”

Gene cascade analysis in human granulosa tumor cells (KGN) following exposure to high levels of free fatty acids and insulin

“…Thought to be key transcription factors for immune cell activation and differentiation, NFAT family members were also suggested to be involved in adipocyte differentiation, modulation of adipokine gene expression, and to participate in glucose and insulin homeostasis [ 105 , 107 – 109 ]. Although NFATC1 was upregulated following stimulation of porcine granulosa cells with a phospholipase C (PLC) activator [ 110 ], the actions mediated by the NFAT family in granulosa cells still remain poorly understood. Functional classification of the 38 downstream effector genes of the NFAT canonical pathway using PANTHER Classification systems showed that the differentially expressed genes were, among others, related to inflammation mediated by chemokine and cytokine signaling pathways, and to the 5HT2 type receptor or serotonin receptor (5-HT2A) signaling pathway which is known to stimulate an increase in intracellular Ca 2+ levels via the PLC/IP3/DAG pathway [ 111 ].…”

Isorhamnetin Protects Zearalenone-induced Damage via the PI3K/Akt Signaling Pathway in Porcine Granulosa Cells

“…These data suggest that isorhamnetin restores GCs proliferation against ZEA-induced apoptosis and subsequently increases the S phase of DNA synthesis via the PI3K/Akt. Healthy follicles have higher concentrations of E2 [14,36]. Thus, the steroidogenesis of GCs is essential for atresia and follicular development.…”

“…GCs were mixed in DMEM/F12 medium supplemented with1% streptomycin-penicillin and 8% fetal bovine serum (FBS) (Serapro, Germany). GCs were cultured at 37°C in 5% CO2 for 36 to 44 h. After, GCs were cultured with isorhamnetin and ZEA, and incubated for 24 h in DMEM/F12 medium containing transferrin (2.5 mg/mL), 0.5% bovine serum albumin (BSA) (w/v), sodium bicarbonate (10 mM), insulin (50 ng/mL), sodium selenite (5 ng/mL), FSH (0.1 IU/mL), penicillin (100 U/mL), streptomycin (100 mg/mL) and nonessential amino acid mix (1×) [14]. GCs were collected for protein and mRNA extraction.…”