乐芮(Luveris)与果纳芬(Gonal-F)对牛未成熟卵母细胞体外发育潜力的影响
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摘要
目的:探讨黄体生成素乐芮药物(Luveris,L)及卵泡刺激素果纳芬药物(Gonal-F,G)对体外未成熟牛卵母细胞成熟培养的影响。方法:购买商品化的牛未成熟卵母细胞2 000枚,分为2组,Luveris组及Gonal-F组,每组1 000枚。每组根据药物浓度0、5、10、20、40IU/mL再分组培养,培养24、48h后,解剖显微镜下评估COCs扩展程度,倒置显微镜下对卵母细胞核成熟(GV,MI,MⅡ)及第一极体排出情况进行评估。结果:Luveris及Gonal-F均随剂量依赖性显著性促进COCs的扩展。L与G培养24h和48h卵母细胞MⅡ成熟率分别是20IU/mL L为33.3%和37.5%,40IU/mL L为33.3%和53.6%;20IU/mL G为15.1%和16.2%;40IU/mL G为38.9%和39.5%。结论:黄体生成素可以像卵泡刺激素一样在卵泡发育早期促进体外牛卵母细胞的成熟发育,可以延迟卵母细胞GVBD。
Objective:To investigate the gonadotrophin drug Luveris(L)and Gonal-F(G)effects on bovine immature oocyte in vitro maturation culture.Method:In total,2 000 bovine cumulus-oocyte complexes(COCs)were purchased commercially and cultured in media with ovulation stimulation drugs Gonal-F and Luveris(0,5,10,20,40 IU/mL),individually,for maturation in vitro.2 000 Oocytes were divided evenly into two groups,Luveris and Gonal-F groups,individually,1 000 Oocytes.After 24 and 48 hculture,cumulus expansion was assessed under dissecting microscopes.Oocyte maturation(GV,MI,MIⅡ)was determined by examining nuclear maturation and polar body formation under inverted microscopes.Results:Luveris and Gonal-F enhanced oocyte maturation in vitro in a dose-dependently manner.Culture media supplemented with L or G significantly improved MⅡ Oocyte maturation rates after culture for 24 hand 48 hin comparison with the control group.Oocyte maturation rates were 33.3% and 37.5%(20 IU/mL),individually,33.3%and 53.6%(40 IU/mL);20 IU/mL G was 15.1%and 16.2%;40 IU/mL G was 38.9% and 39.5%,respectively.Conclusion:LH(Luveris)can promote prophase bovine immature oocytes developmental competence in vitro,which is mostly likely stimulated by FSH(Gonal-F).Furthermore,LH can delay oocytes GVBD that is very useful for IVF clinical medication guidance.
Objective:To investigate the gonadotrophin drug Luveris(L)and Gonal-F(G)effects on bovine immature oocyte in vitro maturation culture.Method:In total,2 000 bovine cumulus-oocyte complexes(COCs)were purchased commercially and cultured in media with ovulation stimulation drugs Gonal-F and Luveris(0,5,10,20,40 IU/mL),individually,for maturation in vitro.2 000 Oocytes were divided evenly into two groups,Luveris and Gonal-F groups,individually,1 000 Oocytes.After 24 and 48 hculture,cumulus expansion was assessed under dissecting microscopes.Oocyte maturation(GV,MI,MIⅡ)was determined by examining nuclear maturation and polar body formation under inverted microscopes.Results:Luveris and Gonal-F enhanced oocyte maturation in vitro in a dose-dependently manner.Culture media supplemented with L or G significantly improved MⅡ Oocyte maturation rates after culture for 24 hand 48 hin comparison with the control group.Oocyte maturation rates were 33.3% and 37.5%(20 IU/mL),individually,33.3%and 53.6%(40 IU/mL);20 IU/mL G was 15.1%and 16.2%;40 IU/mL G was 38.9% and 39.5%,respectively.Conclusion:LH(Luveris)can promote prophase bovine immature oocytes developmental competence in vitro,which is mostly likely stimulated by FSH(Gonal-F).Furthermore,LH can delay oocytes GVBD that is very useful for IVF clinical medication guidance.
引文
1谢幸,荀文丽.妇产科学[M].8版.北京:人民卫生出版社,2014:24-26.
2邓宇,尹敏娜,梁佩玲,等.长方案添加黄体生成素对体外受精-胚胎移植结局的影响[J].南方医科大学学报,2017,37(11):1501-1505.
3 Xiao X,Zi XD,Niu HR,et al.Effect of addition of FSH,LH and proteasome inhibitor MG132to in vitro maturation medium on the developmental competence of yak(Bos grunniens)oocytes[J].Reprod Biol Endocrinol.2014,22(12):30.
4 Rahman A,Francomano D,Sagnella F,et al.The effect on clinical results of adding recombinant LH in late phase of ovarian stimulation of patients with repeated implantation failure:apilot study[J].Eur Rev Med Pharmacol Sci,2017,21(23):5485-5490.
5 Lee HS,Seo YI,Yin XJ,et al.Effect of follicle stimulation hormone and luteinizing hormone on cumulus cell expansion and in vitro nuclear maturation of canine oocytes[J].Reprod Domest Anim,2007,42:561-565.
6 Sha W,Xu BZ,Li M,et al.Effect of gonadotropins on oocyte maturation in vitro:an animalmodel[J].Fertil Steril,2010,93:1650-1661.
7 Dinopoulou V,Drakakis P,Kefala S,et al.Effect of recombinant-LH and hCG in the absence of FSH on in vitro maturation(IVM)fertilization and early embryonic development of mouse germinal vesicle(GV)-stage oocytes[J].Reprod Biol,2016,16(2):138-140.
8 Egbert JR,Yee SP,Jaffe LA.Luteinizing hormone signaling phosphorylates and activates the cyclic GMPphosphodiesterase PDE5in mouse ovarian follicles,contributing an additional component to the hormonally induced decrease in cyclic GMP that reinitiates meiosis[J].Dev Biol,2018,435(1):6-14.
9 Firmani LD,Uliasz TF,Mehlmann LM.The switch from cAMP-independent to cAMP-dependent arrest of meiotic prophase is associated with coordinated GPR3and CDK1expression in mouse oocytes[J].Dev Biol,2017,434(1):196-205.
10 Durnerin CI,Erb K,Fleming R,et al.Luveris Pretreatment Group.Effects of recombinant LH treatment on folliculogenesis and respon-siveness to FSH stimulation[J].Hum Reprod 2008,23,421-426.
11 Shahrokh Tehraninejad E,Farshbaf Taghinejad M,Hossein Rashidi B,et al.Controlled ovarian stimulation with r-FSH plus r-LH vs.HMG plus r-FSH in patients candidate for IVF/ICSI cycles:An RCT[J].Int J Reprod Biomed(Yazd),2017,15(7):435-440.
12 Nivet AL,Vigneault C,Blondin P,et al.Influence of luteinizing hormone support on granulosa cells transcriptome in cattle[J].Anim Sci J,2018,89(1):21-30.
2邓宇,尹敏娜,梁佩玲,等.长方案添加黄体生成素对体外受精-胚胎移植结局的影响[J].南方医科大学学报,2017,37(11):1501-1505.
3 Xiao X,Zi XD,Niu HR,et al.Effect of addition of FSH,LH and proteasome inhibitor MG132to in vitro maturation medium on the developmental competence of yak(Bos grunniens)oocytes[J].Reprod Biol Endocrinol.2014,22(12):30.
4 Rahman A,Francomano D,Sagnella F,et al.The effect on clinical results of adding recombinant LH in late phase of ovarian stimulation of patients with repeated implantation failure:apilot study[J].Eur Rev Med Pharmacol Sci,2017,21(23):5485-5490.
5 Lee HS,Seo YI,Yin XJ,et al.Effect of follicle stimulation hormone and luteinizing hormone on cumulus cell expansion and in vitro nuclear maturation of canine oocytes[J].Reprod Domest Anim,2007,42:561-565.
6 Sha W,Xu BZ,Li M,et al.Effect of gonadotropins on oocyte maturation in vitro:an animalmodel[J].Fertil Steril,2010,93:1650-1661.
7 Dinopoulou V,Drakakis P,Kefala S,et al.Effect of recombinant-LH and hCG in the absence of FSH on in vitro maturation(IVM)fertilization and early embryonic development of mouse germinal vesicle(GV)-stage oocytes[J].Reprod Biol,2016,16(2):138-140.
8 Egbert JR,Yee SP,Jaffe LA.Luteinizing hormone signaling phosphorylates and activates the cyclic GMPphosphodiesterase PDE5in mouse ovarian follicles,contributing an additional component to the hormonally induced decrease in cyclic GMP that reinitiates meiosis[J].Dev Biol,2018,435(1):6-14.
9 Firmani LD,Uliasz TF,Mehlmann LM.The switch from cAMP-independent to cAMP-dependent arrest of meiotic prophase is associated with coordinated GPR3and CDK1expression in mouse oocytes[J].Dev Biol,2017,434(1):196-205.
10 Durnerin CI,Erb K,Fleming R,et al.Luveris Pretreatment Group.Effects of recombinant LH treatment on folliculogenesis and respon-siveness to FSH stimulation[J].Hum Reprod 2008,23,421-426.
11 Shahrokh Tehraninejad E,Farshbaf Taghinejad M,Hossein Rashidi B,et al.Controlled ovarian stimulation with r-FSH plus r-LH vs.HMG plus r-FSH in patients candidate for IVF/ICSI cycles:An RCT[J].Int J Reprod Biomed(Yazd),2017,15(7):435-440.
12 Nivet AL,Vigneault C,Blondin P,et al.Influence of luteinizing hormone support on granulosa cells transcriptome in cattle[J].Anim Sci J,2018,89(1):21-30.