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Research Paper Volume 11, Issue 19 pp 8362-8373
Mogroside V protects porcine oocytes from in vitro ageing by reducing oxidative stress through SIRT1 upregulation
Relevance score: 11.867878Junyu Nie, Lumin Sui, Huiting Zhang, Hengye Zhang, Ke Yan, Xiaogan Yang, Shengsheng Lu, Kehuan Lu, Xingwei Liang
Keywords: mogroside V, oocyte ageing in vitro, ROS, SIRT1, embryo development
Published in Aging on October 6, 2019
Mogroside V alleviates the deterioration in oocyte quality during in vitro ageing. After in vitro maturation for 44 h, oocytes that extruded the first polar body were continuously cultured in vitro with or without MV (25, 50 and 100 μM) for 24 h. (A) Views of oocytes after activation. (B) Activation rates of oocytes in the fresh, aged, and aged+MV groups. (C) Representative images of blastocysts from fresh, aged and aged+MV oocytes. (D) Blastocyst formation rates of fresh, aged and aged+MV oocytes. MV, mogroside V; Scale bar = 200 μm. The data are presented as the mean ± SEM of at least three independent experiments. * P < 0.05, *** P < 0.001, **** P < 0.0001.
Effects of mogroside V on the ROS level and SIRT1 expression in oocytes during in vitro ageing. After in vitro maturation for 44 h, oocytes that extruded the first polar body were continuously cultured in vitro with or without MV for 24 h. (A) Representative images of fresh, aged and aged+MV oocytes stained with DCFH-DA. (B) Quantitative analysis of ROS fluorescence intensity in fresh, aged and aged+MV oocytes. (C) The mRNA expression of SIRT1 in fresh, aged and aged+MV oocytes. The data are presented as the mean ± SEM of at least three independent experiments. MV, mogroside V; Scale bar = 100 μm. * P< 0.05, ** P < 0.01, **** P< 0.0001.
Effects of mogroside V on spindle formation and chromosome alignment in oocytes during in vitro ageing. After in vitro maturation for 44 h, oocytes that extruded the first polar body were continuously cultured in vitro with or without MV for 24 h. (A) Representative images of spindle morphologies and chromosome alignment in fresh, aged and aged+MV oocytes. (B) The percentage of aberrant spindle formation in fresh, aged and aged+MV oocytes. (C) The percentage of misaligned chromosomes in fresh, aged and aged+MV oocytes. The data are presented as the mean ± SEM of at least three independent experiments. MV, mogroside V; Scale bar = 7.5 μm. ** P<0.01, *** P< 0.001.
Effects of mogroside V on mitochondrial and ATP contents in oocytes during in vitro ageing. After in vitro maturation for 44 h, oocytes that extruded the first polar body were continuously cultured in vitro with or without MV for 24 h. (A) Representative images of fresh, aged and aged+MV oocytes stained with MitoTracker™ Orange CMTMRos. (B) Quantitative analysis of the mitochondrial content. (C) Quantitative analysis of the ATP content. The data are presented as the mean ± SEM of at least three independent experiments. MV, mogroside V; Scale bar = 50 μm. * P < 0.05, *** P < 0.001, **** P < 0.0001.
Effect of mogroside V on the mitochondrial membrane potential in oocytes during in vitro ageing. After in vitro maturation for 44 h, oocytes that extruded the first polar body were continuously cultured in vitro with or without MV for 24 h. (A) Representative images of fresh, aged and aged+MV oocytes stained with JC-1. (B) Quantitative analysis of JC-1 fluorescence intensity. The data are presented as the mean ± SEM of at least three independent experiments. Mogroside V, MV; Scale bar = 200 μm. * P< 0.05, **** P< 0.0001.
Effect of mogroside V on early apoptosis in oocytes during in vitro ageing. After in vitro maturation for 44 h, oocytes that extruded the first polar body were continuously cultured in vitro with or without MV for 24 h. (A) Representative images of fresh, aged and aged+MV oocytes stained with Annexin-V. (B) Quantitative analysis of Annexin-V fluorescence intensity. The data are presented as the mean ± SEM of at least three independent experiments. Mogroside V, MV; Scale bar = 50 μm. *** P < 0.001, **** P < 0.0001.
Inhibition of SIRT1 abolishes the protective effect of mogroside V on oocytes during in vitro ageing. After in vitro maturation for 44 h, oocytes that extruded the first polar body were continuously cultured in vitro with MV or MV plus the SIRT1 inhibitor EX527 for 24 h. (A) Representative images of aged, aged+MV and aged+MV+EX-527 oocytes stained with SIRT1 antibody. (B) Quantitative analysis of SIRT1 fluorescence intensity. (C) Representative images of aged, aged+MV and aged+MV+EX-527 oocytes stained with DCFH-DA. (D) Quantitative analysis of ROS fluorescence intensity. (E) Representative images of spindle morphologies and chromosome alignment in aged, aged+MV and aged+MV+EX-527 oocytes. (F) Quantitative analysis of the abnormal spindle formation rate. (G) Quantitative analysis of the misaligned chromosome rate. The data are presented as the mean ± SEM of at least three independent experiments. Mogroside V, MV; Scale bar, A = 50 μm, C = 200 μm, E = 5 μm. * P < 0.05, ** P <0.01, **** P < 0.0001.