甘氨酸对小鼠早期胚胎发育影响的剂量效应研究
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摘要
为更好地了解甘氨酸对胚胎发育的影响,本研究利用小鼠胚胎体外培养方法,分析了早期胚胎在含有不同浓度甘氨酸的KSOM培养液中的发育,并结合定量PCR方法分析了5个囊胚期重要发育基因的表达。结果表明,甘氨酸浓度达到2.5μg·mL~(-1)时,小鼠囊胚的形成不受影响。但当浓度达到5μg·mL~(-1)及以上时,胚胎发育明显受到抑制。对囊胚期5个基因表达的分析表明,与对照组相比,甘氨酸可以显著改变实验组基因的表达。结果表明甘氨酸对小鼠胚胎发育的影响存在着浓度依赖性,高浓度下会显著抑制胚胎发育,并且会改变胚胎发育中重要基因的表达。
To better understand the effect of glycine on embryonic development, the development profile of mouse's early embryos in KSOM medium with different concentrations of glycine was analyzed, and the expression of 5 important developmental genes at blastocyst stage was investigated by quantitative PCR. The results showed that the formation of blastocysts in the group of 2.5 μg · mL~(-1) glycine was not different to control group. But when the concentration reached 5 μg · mL~(-1) and above,the embryo development is obviously inhibited. The analysis on the expression of 5 genes in blastocyst showed that glycine could significantly alter their expression level in the experimental group compared with the control group. All above indicate that effects of glycine on mouse embryonic development are dose-dependent.
To better understand the effect of glycine on embryonic development, the development profile of mouse's early embryos in KSOM medium with different concentrations of glycine was analyzed, and the expression of 5 important developmental genes at blastocyst stage was investigated by quantitative PCR. The results showed that the formation of blastocysts in the group of 2.5 μg · mL~(-1) glycine was not different to control group. But when the concentration reached 5 μg · mL~(-1) and above,the embryo development is obviously inhibited. The analysis on the expression of 5 genes in blastocyst showed that glycine could significantly alter their expression level in the experimental group compared with the control group. All above indicate that effects of glycine on mouse embryonic development are dose-dependent.
引文
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[18]REDEL B K,SPATE L D,LEE K,et al.Glycine supplementation in vitro enhances porcine preimplantation embryo cell number and decreases apoptosis but dies lead to live births[J].Molecular Reproduction and Development,2016,83:246-258.
[19]PIOTR K,MARIUSZ P K,AGNIESZKA W.Prostaglandin F2αpromotes angiogenesis and embryo-maternal interactions during implantation[J].Society for Reproduction and Fertility,2016,151:539-552.
[20]高国龙.卵巢因子对小鼠卵母细胞发育的影响[J].湖北农业科学,2012,51(6):1085-1088.
[21]NOEL S D,ABREU A P,XU S Y,et al.TACR3 mutations disrupt NK3R function through distinct mechanisms in GnRH-deficient patients[J].FASEB Journal,2014,28(4):1924-1937.
[22]白小双.体外受精-胚胎移植过程中卵泡液血小板激活因子的研究[D].石家庄:河北医科大学,2014:26-28.
[23]YI K H,YU M,WU L B,et al.Effects of urotensin IIon functional activity of late endothelial progenitor cells[J].Peptides,2012,33(1):87-91.
[2]LIU L,FU T,XU X,et al.Tracing the nitrogen metabolites of glycine using 15Nglycine and mass spectrometry[J].Rapid Communicaations in Mass Spectrometry,2015,29:645-653.
[3]赵雪,陆大祥,戚仁斌,等.甘氨酸受体在甘氨酸抗心肌细胞缺氧/复氧损伤中的作用机制研究[J].中国病理生理杂志,2006,22(11):2113-2118.
[4]ALARCON-AGUILAR F J,ALMANZA-PEREZ J,BLANCAS G,et al.Glycine regulates the production of pro-inflammatory cytokines in lean and monosodium glutamate-obese mice[J].European Journal of Pharmacology,2008,599(1/3):152.
[5]廉洪,陈欢,杜景霞,等.甘氨酸对离体缺血再灌注大鼠心脏的保护作用[J].华中科技大学学报(医学版),2006,35(6):728-730.
[6]BROOKS R F.Continuous protein synthesis is required to maintain the probability of entry into S phase[J].Cell,1977,12:311-317.
[7]LAWRENCE JC,ABRAHAM RT.PHAS/4E-BPs as regulators of mRNA translation and cell proliferation[J].Trends in Biochemical Sciences,1997,22:345-349.
[8]GEORGE T,MICHAEL N H.Tor signalling and control of cell growth[J].Current Opinion in Cell Biology,1997,9:782-787.
[9]MITO T,YOSHIOKA K,YAMASHITA S,et al.Glucose and glycine synergistically enhance the in vitro development of porcine blastocysts in a chemically defined medium[J].Reproduction Fertility and Development,2012,24(3):443-450.
[10]RIEGER D,LOSKUTOFF N M,BETTERIDGE K J.Developmentally related changes in the uptake and metabolism of glucose,glutamine,and pyruvate by cattle embryos produced in vitro[J].Reproduction,Fertility and Development,1992,4:547-557.
[11]XU J,SINCLAIR K D.One-carbon metabolism and epigenetic regulation of embryo development[J].Reproduction Fertility and Development,2015,27(4,SI):667-676.
[12]LANE M,GARDNER D K.Mitochondrial malate-aspartate shuttle regulates mouse embryo nutrient consumption[J].The Journal of Biological Chemistry,2005,280:18361-18367.
[13]PATRICK M,ANN E S.Exogenous amino acids regulate trophectoderm differentiation in the mouse blastocyst through an mTOR-dependent pathway[J].Developmental Biology,2001,240:182-193.
[14]BALTZ J M,ZHOU C X.Cell volume regulation in mammalian oocytes and preimplantation embryos[J].Molecular Reproduction and Development,2012,79(12):821-831.
[15]BALTZ JM.Osmoregulation and cell volume regulation in the preimplantation embryo[J].Curr Top Dev Biol 2001,52:55-106.
[16]MIYOSHI K,MIZOBE Y.Osmolarity-and Stage-Dependent effects of glycine on parthenogenetic development of pig oocytes[J].Journal of Reproduction and Development,2014,60(5):349-354.
[17]纳吉A,玛丽娜·格特森斯坦,克里斯蒂娜·文特斯藤,等编.孙青原,陈大元译[M].第3版.北京:化学工业出版社,2005:146-148.
[18]REDEL B K,SPATE L D,LEE K,et al.Glycine supplementation in vitro enhances porcine preimplantation embryo cell number and decreases apoptosis but dies lead to live births[J].Molecular Reproduction and Development,2016,83:246-258.
[19]PIOTR K,MARIUSZ P K,AGNIESZKA W.Prostaglandin F2αpromotes angiogenesis and embryo-maternal interactions during implantation[J].Society for Reproduction and Fertility,2016,151:539-552.
[20]高国龙.卵巢因子对小鼠卵母细胞发育的影响[J].湖北农业科学,2012,51(6):1085-1088.
[21]NOEL S D,ABREU A P,XU S Y,et al.TACR3 mutations disrupt NK3R function through distinct mechanisms in GnRH-deficient patients[J].FASEB Journal,2014,28(4):1924-1937.
[22]白小双.体外受精-胚胎移植过程中卵泡液血小板激活因子的研究[D].石家庄:河北医科大学,2014:26-28.
[23]YI K H,YU M,WU L B,et al.Effects of urotensin IIon functional activity of late endothelial progenitor cells[J].Peptides,2012,33(1):87-91.