In vitro influence of selenium on the proliferation of and steroidogenesis in goat luteinized granulosa cells

Yao, Xiaolei; EI-Samahy, M.A.; Fan, Lijie; Zheng, Linfeng; Jin, Yuyue; Pang, Jing; Zhang, Guomin; Liu, Zifei; Wang, Feng

doi:10.1016/j.theriogenology.2018.03.014

Cited by 39 publications

(34 citation statements)

References 34 publications

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“…Goat ovaries in follicular phase were collected from a local abattoir (Zhenjiang, Jiangsu, China) and brought to the lab within 2 hours in 0.9% NaCl solution (30‐35℃). GCs were collected from the follicular fluid of healthy follicles (2‐5 mm in diameter) as described by Yao et al Cell viability was > 90% as determined by trypan blue dye. The freshly isolated goat GCs were seeded at a density of 5×10 4 cells/cm 2 , and incubated in basic culture medium (BCM; DMEM‐F12 including 10% FBS, 100 IU/mL of penicillin, and 100 μg/mL of streptomycin) at 37℃ with 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

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Effects of LPS on the accumulation of lipid droplets, proliferation, and steroidogenesis in goat luteinized granulosa cells

Zhang

Wang

et al. 2019

J Biochem & Molecular Tox

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Lipopolysaccharide (LPS) can cause ovarian dysfunction and infertility in mammals. The purpose of this study was to investigate the effects of LPS on the accumulation of lipid droplets (LDs), proliferation, and steroidogenesis in goat luteinized granulosa cells (LGCs). GCs isolated from the ovarian follicles were spontaneously luteinized under media with fetal bovine serum, resulting in increased progesterone and shifted shape from spherical to star with multiple prolongations. Then, LGCs were treated with LPS (0-10 μg/mL) for 0-48 hours. Oil Red O staining was performed to observe LDs accumulation and commercial kit was applied to detect intracellular triglyceride (TG) content. The cell proliferation were detected by cell counting kit-8. Expressions of cellcycle-related genes were determined by real-time polymerase chain reaction. Estradiol (E 2 ) and progesterone (P 4 ) from cell supernatants were determined by enzyme-linked immunosorbent assay, and expressions of STAR, P450scc, 3β-hydroxysteroid dehydrogenase (3β-HSD) and CYP19A1 were detected by Western blot. Results showed that LPS treatment significantly increased LDs accumulation after 24 hours, and 5 μg/mL LPS increased TG content (P < 0.05). LPS treatment for 24 hours stimulated the LGCs activities (P<0.05), which was confirmed by the increases in the expressions of proliferating cell nuclear antigen (PCNA), cyclinB1 and cyclinD1, while 48 hours treatment had no effect. LPS treatment suppressed E 2 and P 4 output of LGCs (P < 0.05). Western blot results showed that 10 μg/mL LPS decreased the protein expression of 3β-HSD in LGCs (P < 0.05). In conclusion, LPS increased LDs accumulation and cell proliferation, and LPS-mediated P 4 reduction could be attributed to the decreased 3β-HSD protein expression, which provide new information for the regulation of ovarian function in goats. HIGHLIGHTSLPS increased the content of lipid droplet in goat luteinized granulosa cells. LPS promoted the proliferation of goat luteinized granulosa cells after treatment for 24 hours. LPS suppressed P4 and E2 output which can be attributed to the decreased 3β-HSD protein expression. K E Y W O R D S cell proliferation, lipid droplet, lipopolysaccharide, luteinized granulosa cells, steroidogenesis J Biochem Mol Toxicol. 2019;33:e22329.wileyonlinelibrary.com/journal/jbt

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Section: Methodsmentioning

confidence: 99%

“…Experiment 1: GCs luteinization. GCs were luteinized as in previous study . Briefly, GCs were allowed to attach in the BCM.…”

Section: Methodsmentioning

confidence: 99%

Effects of LPS on the accumulation of lipid droplets, proliferation, and steroidogenesis in goat luteinized granulosa cells

Zhang

Wang

et al. 2019

J Biochem & Molecular Tox

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“…PI3K (phosphatidylinositide 3-kinases) pathway has been implicated to play an important role in integration of several known and unknown factors principally associated with maintenance of primordial follicle dynamics [70]. Interestingly, in a recent in vitro cell culture study, it was shown that Se supplementation can stimulate the proliferation of caprine luteinized-granulosa cells and steroidogenesis potentially via modulating PI3K/Akt and AMP-activated protein kinase (AMPK) pathways and expression of downstream genes related to cell cycle and steroidogenesis, and improving the cellular oxidative status [45]. In addition, a recent pilot study [28] on bovine ovaries has shown that Se was consistently localized in the granulosa and theca cells of healthy and atretic follicles.…”

Section: Effect Of Se On Follicle Quantity and Apoptosis In Ovarian Tmentioning

confidence: 99%

“…As an inhibitor of cyclin-dependent kinases (CDKs), p21 has been shown to impair G1-to-S-phase cell cycle progression via modulating cell cycle check-points [113]. In a recent in vitro study [45], it was shown that Se supplementation (5 ng/mL) improved proliferative capacity of luteinized granulosa cells, potentially via downregulation of P21 expression both at mRNA and protein levels. Similarly, Se-deficient and Se-excess diets also resulted in cell cycle arrest during spermatogenesis in mice, potentially via elevated expression of p21 and decreased expression of CDC 2 and Cyclin B1 [113].…”

Section: Expression Of Gpx1 Gpx3 Gpx4 Selenof Bcl-2 and P21mentioning

confidence: 99%

“…It was shown that Se was implicated in regulating the physiology of uterine smooth muscles potentially via modulating the expression of Selenon, Selenot, and Selenow [41,42], and the G protein Rho (RhoA)/Rho kinase (ROCK) signaling pathway in mice [43]. However, only recently have researchers paid attention to potential role of Se in ovarian physiology and embryo development [9,28,[44][45][46][47][48][49][50]. Interestingly, when exploring the mainstream literature, with the exclusion of one study reporting the beneficial effects of Se supplementation on sperm parameters in aging mice [51], no study has yet elucidated how Se might ameliorate the reproductive function in aging mammalian models, let alone the aged females.…”

Section: Introductionmentioning

confidence: 99%

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Dietary Selenium Supplementation Ameliorates Female Reproductive Efficiency in Aging Mice

et al. 2019

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Female reproductive (ovarian) aging is distinctively characterized by a markedly reduced reproductive function due to a remarkable decline in quality and quantity of follicles and oocytes. Selenium (Se) has been implicated in playing many important biological roles in male fertility and reproduction; however, its potential roles in female reproduction, particularly in aging subjects, remain poorly elucidated. Therefore, in the current study we used a murine model of female reproductive aging and elucidated how different Se-levels might affect the reproductive efficiency in aging females. Our results showed that at the end of an 8-week dietary trial, whole-blood Se concentration and blood total antioxidant capacity (TAOC) were significantly reduced in Se-deficient (0.08 mg Se/kg; Se-D) mice, whereas both of these biomarkers were significantly higher in inorganic (0.33 mg/kg; ISe-S) and organic (0.33 mg/kg; OSe-S) Se-supplemented groups. Similarly, compared to the Se-D group, Se supplementation significantly ameliorated the maintenance of follicles and reduced the rate of apoptosis in ovaries. Meanwhile, the rate of in vitro-produced embryos resulting from germinal vesicle (GV) oocytes was also significantly improved in Se-supplemented (ISe-S and OSe-S) groups compared to the Se-D mice, in which none of the embryos developed to the hatched blastocyst stage. RT-qPCR results revealed that mRNA expression of Gpx1, Gpx3, Gpx4, Selenof, p21, and Bcl-2 genes in ovaries of aging mice was differentially modulated by dietary Se levels. A considerably higher mRNA expression of Gpx1, Gpx3, Gpx4, and Selenof was observed in Se-supplemented groups compared to the Se-D group. Similarly, mRNA expression of Bcl-2 and p21 was significantly lower in Se-supplemented groups. Immunohistochemical assay also revealed a significantly higher expression of GPX4 in Se-supplemented mice. Our results reasonably indicate that Se deficiency (or marginal levels) can negatively impact the fertility and reproduction in females, particularly those of an advancing age, and that the Se supplementation (inorganic and organic) can substantiate ovarian function and overall reproductive efficiency in aging females.

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Neddylation inactivation affects cell cycle and apoptosis in sheep follicular granulosa cells

Qin

Dang

Yang

et al. 2022

Journal Cellular Physiology

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Protein neddylation inactivation is a novel topic in cancer research. However, there are few studies on the mechanism of neddylation underlying the development of sheep follicular granulosa cells (GCs). In this study, the development of follicular GCs in sheep was inactivated by MLN4924, a neddylation‐specific inhibitor, which significantly attenuated the proliferation and cell index of sheep follicular GCs. Further, the inactivation of neddylation by MLN4924 caused the accumulation of the cullin ring ligase (CRLs) substrates Wee1 and c‐Myc, which could upregulate NOXA protein expression. Meanwhile, the B‐cell lymphoma/leukemia 2 (BCL2) family members Bcl‐2 and MCL‐1 were downregulated, subsequently inducing apoptosis in follicular GCs of sheep. Increasing Wee1 levels caused G2/M‐phase arrest. The effects of neddylation inactivation on Akt, the JAK2/STAT3 signaling pathway, and Forkhead box class O(FOXO) family members were evaluated. Neddylation inactivation by MLN4924 increased the levels of phospho‐Akt, JAK2, phospho‐STAT3, and FOXO1 (p < 0.05) and decreased the levels of phospho‐FOXO3a and STAT3 (p < 0.05). In addition, MLN4924 could alter the mitochondrial morphology of GCs, increase cellular glucose utilization and lactate production, increase reactive oxygen species (ROS) generation, and promote sheep follicular GCs glycolysis, thus causing changes in mitochondrial functions. Together, these findings point to an unrecognized role of neddylation in regulating follicular GCs proliferation in sheep.

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In vitro influence of selenium on the proliferation of and steroidogenesis in goat luteinized granulosa cells

Cited by 39 publications

References 34 publications

Effects of LPS on the accumulation of lipid droplets, proliferation, and steroidogenesis in goat luteinized granulosa cells

Effects of LPS on the accumulation of lipid droplets, proliferation, and steroidogenesis in goat luteinized granulosa cells

Dietary Selenium Supplementation Ameliorates Female Reproductive Efficiency in Aging Mice

Neddylation inactivation affects cell cycle and apoptosis in sheep follicular granulosa cells

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