不同促排卵方案对小鼠卵巢颗粒细胞凋亡的影响
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摘要
目的研究不同促排卵方案对小鼠卵巢颗粒细胞Fas凋亡通路的影响。方法将雌性KM小鼠随机分为4组:孕马血清促排卵组(PMSG组)、促性腺激素释放激素激动剂组(GnRH-a组)、促性腺激素释放激素拮抗剂组(GnRH-ant组)、自然排卵组(空白对照组),每组10只。取排卵期卵巢组织,采用免疫组化检测卵巢组织凋亡因子Fas、FasL、Caspase8、Caspase3的表达情况。结果 4组小鼠卵巢颗粒细胞均有Fas、FasL、Caspase8、Caspase3的表达;经不同促排卵方案干预后,小鼠卵巢颗粒细胞免疫组化Fas、FasL、Caspase8、Caspase3表达水平总体趋势为:GnRH-a组/GnRH-ant组>空白对照组>PMSG组。其中,PMSG组小鼠卵巢颗粒细胞Fas、FasL、Caspase8、Caspase3表达低于空白对照组,但差异无统计学意义(P>0.05);GnRH-a组与GnRH-ant组小鼠卵巢颗粒细胞Fas、FasL、Caspase8、Caspase3表达显著高于空白对照组(P<0.05)。结论不同促排卵方案对小鼠卵巢颗粒细胞Fas凋亡通路的影响不同,PMSG方案可能抑制颗粒细胞凋亡,GnRH-a方案与GnRH-ant方案可能促进颗粒细胞凋亡。
Objective:To study the effects of different ovulation induction protocols on Fas apoptotic pathway of ovarian granulosa cells in mice.Methods: The female SPF mice were randomly divided into 4 groups:pregnant mare serum gonadotropin group(PMSG group),GnRH agonist group(GnRH-agroup),GnRH antagonist group(GnRH-ant group)and natural ovulation group(blank control group),10 mice in each group.The ovarian tissues were collected,and the expressions of Fas,FasL,Caspase8 and Caspase3 in ovarian tissue were detected by immunohistochemistry.Results:The expressions of Fas,FasL,Caspase8 and Caspase3 were detectable in the ovarian granulosa cells of the four groups.The general trend of immunohistochemical expression of Fas,FasL,Caspase 8 and Caspase 3 in mouse ovarian granulosa cells was as follows:GnRH-agroup/GnRH-ant group>blank control group>PMSG group.The expression of Fas,FasL,Caspase8 and Caspase3 in ovarian granulosa cells of PMSG group was lower than that of blank control group,but the difference was not significant(P>0.05).The expressions of Fas,FasL,Caspase8 and Caspase3 in ovarian granulosa cells of GnRH-agroup and GnRH-ant group were significantly higher than those in the blank control group(P<0.05).Conclusions:Different ovulation induction protocols have different effects on Fas apoptotic pathway in ovarian granulosa cells of mice.PMSG protocol inhibits granulosa cell apoptosis,and GnRH-aprotocol and GnRH-ant protocol promote granulosa cell apoptosis.
Objective:To study the effects of different ovulation induction protocols on Fas apoptotic pathway of ovarian granulosa cells in mice.Methods: The female SPF mice were randomly divided into 4 groups:pregnant mare serum gonadotropin group(PMSG group),GnRH agonist group(GnRH-agroup),GnRH antagonist group(GnRH-ant group)and natural ovulation group(blank control group),10 mice in each group.The ovarian tissues were collected,and the expressions of Fas,FasL,Caspase8 and Caspase3 in ovarian tissue were detected by immunohistochemistry.Results:The expressions of Fas,FasL,Caspase8 and Caspase3 were detectable in the ovarian granulosa cells of the four groups.The general trend of immunohistochemical expression of Fas,FasL,Caspase 8 and Caspase 3 in mouse ovarian granulosa cells was as follows:GnRH-agroup/GnRH-ant group>blank control group>PMSG group.The expression of Fas,FasL,Caspase8 and Caspase3 in ovarian granulosa cells of PMSG group was lower than that of blank control group,but the difference was not significant(P>0.05).The expressions of Fas,FasL,Caspase8 and Caspase3 in ovarian granulosa cells of GnRH-agroup and GnRH-ant group were significantly higher than those in the blank control group(P<0.05).Conclusions:Different ovulation induction protocols have different effects on Fas apoptotic pathway in ovarian granulosa cells of mice.PMSG protocol inhibits granulosa cell apoptosis,and GnRH-aprotocol and GnRH-ant protocol promote granulosa cell apoptosis.
引文
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[12]Lee P,Sata M,Lefer DJ,et al.Fas pathway is a critical mediator of cardiac myocyte death and MI during ischemiareperfusion in vivo[J].Am J Physiol Heart Circ Physiol,2003,284:H456-463.
[13]Li HZ,Han LP,Jiang CM,et al.Effect of dopamine receptor1on apoptosis of cultured neonatal rat cardiomyocytes in simulated ischaemia/reperfusion[J].Basic Clin Pharmacol Toxicol,2008,102:329-336.
[14]Hussein MR.Apoptosis in the ovary:molecular mechanisms[J].Hum Reprod Update,2005,11:162-177.
[15]Zhao S,Saito H,Wang X,et al.Effects of gonadotropinreleasing hormone agonist on the incidence of apoptosis in porcine and human granulosa cells[J].Gynecol ObstetIInvest,2000,49:52-56.
[16]Takekida S,Deguchi J,Samoto T,et al.Comparative analysis of the effects of gonadotropin-releasing hormone agonist on the proliferative activity,apoptosis,and steroidogenesis in cultured porcine granulosa cells at varying stages of follicular growth[J].Endocrine,2000,12:61-67.
[17]Giampietro F,Sancilio S,Tiboni GM,et al.Levels of apoptosis in human granulosa cells seem to be comparable after therapy with a gonadotropin-releasing hormone agonist or antagonist[J].Fertil Steril,2006,85:412-419.
[18]Hassun Filho PA,Ribeiro MA,da Rocha AM,et al.Effect of GnRH down-regulation on cumulus cell viability and apoptosis as measured by fluorescence-activated cell sorting[J].J Assist Reprod Genet,2008,25:467-471.
[19]Chun SY,Billig H,Tilly JL,et al.Gonadotropin suppression of apoptosis in cultured preovulatory follicles:mediatory role of endogenous insulin-like growth factor I[J].Endocrinology,1994,135:1845-1853.
[20]Anderiesz C,Ferraretti A,Magli C,et al.Effect of recombinant human gonadotrophins on human,bovine and murine oocyte meiosis,fertilization and embryonic development in vitro[J].Hum Reprod,2000,15:1140-1148.
[21]孙玉菡,丘彦,幸贵邦,等.抑制素A及促性腺激素对小鼠未成熟卵母细胞体外成熟的影响[J].重庆医科大学学报,2010,35:542-544.
[2]乔杰,张曜耀,李蓉.重视促排卵的质量与安全性[J].中国实用妇科与产科杂志,2015,31:13-17.
[3]Ruan HC,Zhu XM,Luo Q,et al.Ovarian stimulation with GnRH agonist,but not GnRH antagonist,partially restores the expression of endometrial integrin beta3and leukaemiainhibitory factor and improves uterine receptivity in mice[J].Hum Reprod,2006,21:2521-2529.
[4]郭新宇,李海霞,谢妍,等.自然排卵、促排卵和超促排卵状态下小鼠卵巢组织中生长分化因子-9(GDF-9)的表达[J].生殖与避孕,2013,33:1-5.
[5]Worku T,Rehman ZU,Talpur HS,et al.MicroRNAs:new insight in modulating follicular atresia:a review[J].Int J Mol Sci,2017,18:E333.
[6]Matsuda F,Inoue N,Manabe N,et al.Follicular growth and atresia in mammalian ovaries:regulation by survival and death of granulosa cells[J].J Reprod Dev,2012,58:44-50.
[7]Tilly JL,Kowalski KI,Johnson AL,et al.Involvement of apoptosis in ovarian follicular atresia and postovulatory regression[J].Endocrinology,1991,129:2799-2801.
[8]Inoue N,Matsuda F,Goto Y,et al.Role of cell-death ligandreceptor system of granulosa cells in selective follicular atresia in porcine ovary[J].J Reprod Dev,2011,57:169-175.
[9]杨晓葵,宋蕊,王树玉.蛋白激酶B在生长分化因子-9抑制卵泡颗粒细胞凋亡中的机制研究[J].生殖医学杂志,2010,19:419-422.
[10]Wallace M,Cottell E,Gibney MJ,et al.An investigation into the relationship between the metabolic profile of follicular fluid,oocyte developmental potential,and implantation outcome[J].Fertil Steril,2012,97:1078-1084.
[11]孙艳,杜生荣,张文昌.人卵泡颗粒细胞的原代培养与鉴定[J].福建医科大学学报,2011,45:118-121.
[12]Lee P,Sata M,Lefer DJ,et al.Fas pathway is a critical mediator of cardiac myocyte death and MI during ischemiareperfusion in vivo[J].Am J Physiol Heart Circ Physiol,2003,284:H456-463.
[13]Li HZ,Han LP,Jiang CM,et al.Effect of dopamine receptor1on apoptosis of cultured neonatal rat cardiomyocytes in simulated ischaemia/reperfusion[J].Basic Clin Pharmacol Toxicol,2008,102:329-336.
[14]Hussein MR.Apoptosis in the ovary:molecular mechanisms[J].Hum Reprod Update,2005,11:162-177.
[15]Zhao S,Saito H,Wang X,et al.Effects of gonadotropinreleasing hormone agonist on the incidence of apoptosis in porcine and human granulosa cells[J].Gynecol ObstetIInvest,2000,49:52-56.
[16]Takekida S,Deguchi J,Samoto T,et al.Comparative analysis of the effects of gonadotropin-releasing hormone agonist on the proliferative activity,apoptosis,and steroidogenesis in cultured porcine granulosa cells at varying stages of follicular growth[J].Endocrine,2000,12:61-67.
[17]Giampietro F,Sancilio S,Tiboni GM,et al.Levels of apoptosis in human granulosa cells seem to be comparable after therapy with a gonadotropin-releasing hormone agonist or antagonist[J].Fertil Steril,2006,85:412-419.
[18]Hassun Filho PA,Ribeiro MA,da Rocha AM,et al.Effect of GnRH down-regulation on cumulus cell viability and apoptosis as measured by fluorescence-activated cell sorting[J].J Assist Reprod Genet,2008,25:467-471.
[19]Chun SY,Billig H,Tilly JL,et al.Gonadotropin suppression of apoptosis in cultured preovulatory follicles:mediatory role of endogenous insulin-like growth factor I[J].Endocrinology,1994,135:1845-1853.
[20]Anderiesz C,Ferraretti A,Magli C,et al.Effect of recombinant human gonadotrophins on human,bovine and murine oocyte meiosis,fertilization and embryonic development in vitro[J].Hum Reprod,2000,15:1140-1148.
[21]孙玉菡,丘彦,幸贵邦,等.抑制素A及促性腺激素对小鼠未成熟卵母细胞体外成熟的影响[J].重庆医科大学学报,2010,35:542-544.