BALB/c小鼠重复药物流产模型的建立及后续妊娠结局的研究
|
摘要
目的：全球每年人工流产人数众多,并且重复流产比率高。重复流产是否对后续妊娠有影响,目前没有明确结论,亟待建立动物模型对妊娠预后进行研究。本实验目的为建立BALB/c小鼠重复药物流产模型,研究小鼠经历了重复流产之后的妊娠结局。
方法：BALB/c小鼠经历2次RU486药物流产之后,观察后续妊娠结局(产仔量,小鼠出生体重等)；检测妊娠中期吸收胎数量,着床点数量以及雌孕激素水平变化。同时取材E13.5胎盘组织,使用Real-time PCR和免疫组化的方法检测与胎盘功能相关基因的表达状况。并且进一步研究经历了重复流产之后的后代F1雄鼠和F1雌鼠生长发育和生殖能力的变化。
结果：经历了重复药物流产的BALB/c小鼠的后续妊娠过程中,自然流产的比率显著升高。E13.5时,实验组妊娠中期E13.5吸收胎比率显著升高,着床点数目和雌孕激素水平与对照组比较无统计学差异.Real-time PCR和免疫组化结果显示,有重复流产经历的小鼠妊娠E13.5的胎盘组织中GR,11β-HSD1/2,SGK1,TF,CD31和VEGF等表达水平降低,表明重复流产小鼠在后续正常妊娠过程中胎盘功能障碍或者胎盘功能不全。重复流产小鼠的F1代肝脏组织中与代谢有关的基因PPARA,PPARG,GR及11β-HSD1的mRNA表达量显著降低。重复流产组F1雄鼠睾丸重量,精子活动能力,精子体外存活时间,交配能力,以及F1雌鼠见栓后妊娠率均显著低于对照组。
结论：本实验成功建立了BALB/c小鼠重复药物流产模型,研究发现有重复流产经历的小鼠在后续妊娠过程中出现高比率自然流产现象。重复流产经历会影响后续妊娠的胎盘功能,F1雄鼠和F1雌鼠的生殖能力。我们认为有重复流产经历的小鼠在后续妊娠过程中出现自然流产,其影响可能涉及后代的生长发育以及成年后与代谢相关的疾病,值得我们进一步研究探索。综上所述,BALB/c小鼠重复流产模型为研究与人类妊娠相关的疾病提供了有益的研究平台,并且有利于研究妇女及其后代子女的健康。
Aim:Medical or surgical abortion is one of the oldest, most commonly practiced, and most controversial induced procedure performed in the world. Repeated abortions account for a large portion of early pregnancy termination. But knowledge is lacking on risks of recurrent pregnancy after abortion, and other effects of abortion on subsequent pregnancy remain an important public health concern. The widely increasing trend of medical abortion and repeated abortions, particularly becoming frequent in an even younger population, critically require risk estimation. In this study, we aimed to establish a mouse model of the repeated medical termination of pregnancy and determine its subsequent outcomes.
Methods:Mifepriston (RU486) medical abortion in BALB/c mice model was established to investigate the impact of medical abortion on subsequent pregnancy, including litter size and newborn birth weight. Pregnant mice were sacrificed to examine the mid-term pregnancy status and investigate the frequency of fetal resorption and placental function gene expression by real-time PCR and immunohistochemistry. Offspring liver mRNA was harvested for real-time PCR to determine the gene expression. Adult males from Fl generations of control and treatment groups were sacrificed to collect the testes and caudal epididymal sperm for further analysis, and investigated sexual function of Fl male. And we also investigated sexual function of female Fl generations.
Results:Mice that previously experienced early medical abortion exhibited spontaneous abortion and pregnancy loss on subsequent pregnancy. Litter size and newborn birth weight of subsequent pregnancy were likewise adversely affected. On midgestation, spontaneous abortion occured during the subsequent pregnancy after medical abortion by RU486, but the level of progesterone and estrogen were no difference compared with control groups. We detected and analyzed metabolic gene expressions in midgestation placental and offspring liver. Medical abortion caused reduced reproductivity and affected placental dysfunction, with downregulation of tissue factor (TF), platelet endothelial cell adhesion molecule-1(CD31), vascular endothelial growth factor (VEGF), especially a panel of genes for proteins involved in metabolic functions relevant for pregnancy, such as 11β-hydroxysteroiddehydrogenase1/2(11β-HSD1/2), serine/threonine protein kinase1(SGK1) and glucocorticoid receptor (GR) expression were reduced. In the offspring, genes involved in lipid metabolism which might enhance key lipid transcription factors (PPARA and PPARG) and GR/11β-HSD1were down-regulated in the liver. We also found that F1female and male reduced their reproduction ability.
Conclusion:We established an inbred model system investigating the impact of repeated first trimester mifepristone induced abortions, which led to subsequent pregnancy loss, affected placental function related gene expression, and F1generation development. This mechanism may involve epigenetic manipulation and merits further study. Thus, the model provides useful means to study the mechanism underlying the above phenomena, which will ultimately benefit the health of women and their children.
方法：BALB/c小鼠经历2次RU486药物流产之后,观察后续妊娠结局(产仔量,小鼠出生体重等)；检测妊娠中期吸收胎数量,着床点数量以及雌孕激素水平变化。同时取材E13.5胎盘组织,使用Real-time PCR和免疫组化的方法检测与胎盘功能相关基因的表达状况。并且进一步研究经历了重复流产之后的后代F1雄鼠和F1雌鼠生长发育和生殖能力的变化。
结果：经历了重复药物流产的BALB/c小鼠的后续妊娠过程中,自然流产的比率显著升高。E13.5时,实验组妊娠中期E13.5吸收胎比率显著升高,着床点数目和雌孕激素水平与对照组比较无统计学差异.Real-time PCR和免疫组化结果显示,有重复流产经历的小鼠妊娠E13.5的胎盘组织中GR,11β-HSD1/2,SGK1,TF,CD31和VEGF等表达水平降低,表明重复流产小鼠在后续正常妊娠过程中胎盘功能障碍或者胎盘功能不全。重复流产小鼠的F1代肝脏组织中与代谢有关的基因PPARA,PPARG,GR及11β-HSD1的mRNA表达量显著降低。重复流产组F1雄鼠睾丸重量,精子活动能力,精子体外存活时间,交配能力,以及F1雌鼠见栓后妊娠率均显著低于对照组。
结论：本实验成功建立了BALB/c小鼠重复药物流产模型,研究发现有重复流产经历的小鼠在后续妊娠过程中出现高比率自然流产现象。重复流产经历会影响后续妊娠的胎盘功能,F1雄鼠和F1雌鼠的生殖能力。我们认为有重复流产经历的小鼠在后续妊娠过程中出现自然流产,其影响可能涉及后代的生长发育以及成年后与代谢相关的疾病,值得我们进一步研究探索。综上所述,BALB/c小鼠重复流产模型为研究与人类妊娠相关的疾病提供了有益的研究平台,并且有利于研究妇女及其后代子女的健康。
Aim:Medical or surgical abortion is one of the oldest, most commonly practiced, and most controversial induced procedure performed in the world. Repeated abortions account for a large portion of early pregnancy termination. But knowledge is lacking on risks of recurrent pregnancy after abortion, and other effects of abortion on subsequent pregnancy remain an important public health concern. The widely increasing trend of medical abortion and repeated abortions, particularly becoming frequent in an even younger population, critically require risk estimation. In this study, we aimed to establish a mouse model of the repeated medical termination of pregnancy and determine its subsequent outcomes.
Methods:Mifepriston (RU486) medical abortion in BALB/c mice model was established to investigate the impact of medical abortion on subsequent pregnancy, including litter size and newborn birth weight. Pregnant mice were sacrificed to examine the mid-term pregnancy status and investigate the frequency of fetal resorption and placental function gene expression by real-time PCR and immunohistochemistry. Offspring liver mRNA was harvested for real-time PCR to determine the gene expression. Adult males from Fl generations of control and treatment groups were sacrificed to collect the testes and caudal epididymal sperm for further analysis, and investigated sexual function of Fl male. And we also investigated sexual function of female Fl generations.
Results:Mice that previously experienced early medical abortion exhibited spontaneous abortion and pregnancy loss on subsequent pregnancy. Litter size and newborn birth weight of subsequent pregnancy were likewise adversely affected. On midgestation, spontaneous abortion occured during the subsequent pregnancy after medical abortion by RU486, but the level of progesterone and estrogen were no difference compared with control groups. We detected and analyzed metabolic gene expressions in midgestation placental and offspring liver. Medical abortion caused reduced reproductivity and affected placental dysfunction, with downregulation of tissue factor (TF), platelet endothelial cell adhesion molecule-1(CD31), vascular endothelial growth factor (VEGF), especially a panel of genes for proteins involved in metabolic functions relevant for pregnancy, such as 11β-hydroxysteroiddehydrogenase1/2(11β-HSD1/2), serine/threonine protein kinase1(SGK1) and glucocorticoid receptor (GR) expression were reduced. In the offspring, genes involved in lipid metabolism which might enhance key lipid transcription factors (PPARA and PPARG) and GR/11β-HSD1were down-regulated in the liver. We also found that F1female and male reduced their reproduction ability.
Conclusion:We established an inbred model system investigating the impact of repeated first trimester mifepristone induced abortions, which led to subsequent pregnancy loss, affected placental function related gene expression, and F1generation development. This mechanism may involve epigenetic manipulation and merits further study. Thus, the model provides useful means to study the mechanism underlying the above phenomena, which will ultimately benefit the health of women and their children.
引文
[1]Who. Safe abortion:technical and policy guidance for health systems (accessed 23 June 2010). http://www.who.int/reproductivehealth/publications/unsafe_abortion/924 1590343/en/[J].2003.
[2]吴尚纯,邱红燕.中国人工流产的现状与对策建议[J].中国医学科学院学报,2010,(5)：479-482.
[3]Shiqian C, Yimin C, Hong W, et al. Study on risk factors of unwanted pregnancy again among the young abortion women in Peking[J]. Maternal and Child Health Care of China,2008, (9):1264-1266.
[4]吴尚纯,邱红燕.中国人工流产的现状与对策建议[J].中国医学科学院学报,2010,(5)：479-482.
[5]郑晓瑛,杨蓉蓉,陈华,等.中国未婚女青年妊娠及流产需要与实现[J].妇女研究论丛,2011,(6)：42-46.
[6]Cheng Y, Gno X, Li Y, et al. Repeat induced abortions and contraceptive practices among unmarried young women seeking an abortion in China[J], International Journal of Gynecology & Obstetrics,2004,87(2):199-202.
[7]Heikinheimo 0, Gissler M, Suhonen S. Age, parity, history of abortion and contraceptive choices affect the risk of repeat abortion[J]. Contraception,2008,78(2):149-154.
[8]Pazol K, Zane S B, Parker W Y, et al. Abortion surveillance--United States,2008[J]. MMWR Surveill Summ,2011,60(15):1-41.
[9]王丽英.人工流产后继发不孕相关因素分析与对策[J].内蒙古中医药,2008,(5)：65-66.
[10]Wu S. Medical abortion in China [J]. J Am Med Womens Assoc,2000,55(3 Suppl):197-199,204.
[11]Creinin M D. Medical abortion regimens:historical context and overview[J]. Am J Obstet Gynecol,2000,183(2 Suppl):S3-S9.
[12]Henshaw S K, Singh S, Haas T. The incidence of abortion worldwide [J]. Int Fam Plann Persp,1999,25(Suppl):S30-S38.
[13]Donaldson K, Briggs J, Mcmaster D. RU 486:an alternative to surgical abortion[J]. J Obstet Gynecol Neonatal Nurs,1994,23(7):555-559.
[14]Westfall J M,O'Brien-Gonzales A, Barley G. Update on early medical and surgical abortion[J]. J Womens Health,1998,7(8):991-995.
[15]张礼婕,刘小平,樊尚荣,等.深圳市人工流产女青少年的性健康状况调查[J].中国全科医学,2004,(15)：1049-1056.
[16]Zhu Q X, Gao E S, Chen A M, et al. Mifepristone-induced abortion and placental complications in subsequent pregnancy[J]. Hum Reprod,2009, 24(2):315-319.
[17]Charles V E, Polis C B, Sridhara S K, et al. Abortion and long-term mental health outcomes:a systematic review of the evidence[J]. Contraception,2008,78(6):436-450.
[18]Gestation, birth-weight, and spontaneous abortion in pregnancy after induced abortion. Report of Collaborative Study by W. H.O. Task Force on Sequelae of Abortion [J]. Lancet,1979,1(8108):142-145.
[19]Atrash H K, Hogue C J. The effect of pregnancy termination on future reproduction[J]. Baillieres Clin Obstet Gynaecol,1990,4(2):391-405.
[20]郑晓瑛,陈功.中国青少年生殖健康可及性调查基础数据报告[J].人口与发展,2010,(3)：2-16.
[21]周亚玲,杨丽华,王秀丽,等.北京市怀柔区妇幼保健院未婚人工流产情况分析[J].中国妇幼保健,2010,(11)：1515-1516.
[22]赵更力,张小松,王临虹,等.中国部分城市流动人口未婚人工流产女青年生殖健康状况分析[J].生殖医学杂志,2005,(5)：16-19.
[23]刘长新.未婚流动人口人工流产相关因素分析[J].中外医疗,2009,(16).
[24]张美萍.青少年生殖健康教育的必要性[J].中国妇幼保健,2009,(23)：3204-3205.
[25]Szekeres-Bartho J, Kinsky R, Chaouat G. The effect of a progesterone-induced immunologic blocking factor on NK-mediated resorption[J]. Am J Reprod Immunol,1990,24(4):105-107.
[26]Szekeres-Bartho J, Par G, Dombay G, et al. The antiabortive effect of progesterone-induced blocking factor in mice is manifested by modulating NK activity[J]. Cell Immunol,1997,177(2):194-199.
[27]Zhang J, Croy B A. Using ultrasonography to define fetal-maternal relationships:moving from humans to mice[J]. Comp Med,2009,59(6): 527-533.
[28]Adamson S L, Lu Y, Whiteley K J, et al. Interactions between trophoblast cells and the maternal and fetal circulation in the mouse placenta[J]. Dev Biol,2002,250(2):358-373.
[29]Lowit A, Bhattacharya S, Bhattacharya S. Obstetric performance following an induced abortion[J]. Best Pract Res Clin Obstet Gynaecol,2010, 24(5):667-682.
[30]Shah P S, Zao J. Induced termination of pregnancy and low birthweight and preterm birth:a systematic review and meta-analyses[J]. BJOG,2009, 116(11):1425-1442.
[31]Sun Y, Che Y, Gao E, et al. Induced abortion and risk of subsequent miscarriage [J]. Int J Epidemiol,2003,32(3):449-454.
[32]Zhou W, Sorensen H T, Olsen J. Induced abortion and low birthweight in the following pregnancy[J]. Int J Epidemiol,2000,29(1):100-106.
[33]Joyce T, Grossman M. The dynamic relationship between low birthweight and induced abortion in New York City. An aggregate time-series analysis [J]. J Health Econ,1990,9(3):273-288.
[34]Shah P S. Parity and low birth weight and preterm birth:a systematic review and meta-analyses[J]. Acta Obstet Gynecol Scand,2010,89(7): 862-875.
[35]Hyatt M A, Budge H, Walker D, et al. Effects of maternal parity and late gestational nutrition on mRNA abundance for growth factors in the liver of postnatal sheep [J]. Am J Physiol Regul Integr Comp Physiol,2007,292(5): R1934-R1942.
[36]Hyatt M A, Keisler D H, Budge H, et al. Maternal parity and its effect on adipose tissue deposition and endocrine sensitivity in the postnatal sheep[J]. J Endocrinol,2010,204(2):173-179.
[37]Lumey L H, Stein A D. Offspring birth weights after maternal intrauterine undernutrition:a comparison within sibships[J]. Am J Epidemiol,1997,146(10):810-819.
[38]Gardner D S, Buttery P J, Daniel Z, et al. Factors affecting birth weight in sheep:maternal environment[J]. Reproduction,2007,133(1): 297-307.
[39]Sun Y, Che Y, Gao E, et al. Induced abortion and risk of subsequent miscarriage [J]. Int J Epidemiol,2003,32(3):449-454.
[40]Zhou W, Olsen J. Are complications after an induced abortion associated with reproductive failures in a subsequent pregnancy?[J]. Acta Obstet Gynecol Scand,2003,82(2):177-181.
[41]Liang H, Gao E S, Chen A M, et al. Mifepristone-induced abortion and vaginal bleeding in subsequent pregnancy[J]. Contraception,2011,84(6): 609-614.
[42]Ahmed A, Singh J, Khan Y, et al. A New Mouse Model to Explore Therapies for Preeclampsia[J]. PLoS One,2010,5(10):e13663.
[43]Zhang J, Sun R, Wei H, et al. Toll-like receptor 3 agonist enhances IFN-gamma and TNF-alpha production by murine uterine NK cells[J]. Int Immunopharmacol,2007,7(5):588-596.
[44]刘智,常才.胎盘功能不足对胎儿的近期和远期影响[J].现代康复,2000,(7)：1110-1111.
[45]Muller M, Renkawitz R. The glucocorticoid receptor[J]. Biochim Biophys Acta,1991,1088(2):171-182.
[46]Tycko B, Efstratiadis A. Genomic imprinting:piece of cake [J]. Nature, 2002,417(6892):913-914.
[47]Redecha P, van Rooijen N, Torry D, et al. Pravastatin prevents miscarriages in mice:role of tissue factor in placental and fetal injury [J]. Blood,2009,113(17):4101-4109.
[48]Osterud B, Bjorklid E. Sources of tissue factor[J]. Semin Thromb Hemost,2006,32(1):11-23.
[49]Redecha P, Tilley R, Tencati M, et al. Tissue factor:a link between C5a and neutrophil activation in antiphospholipid antibody induced fetal injury [J]. Blood,2007,110(7):2423-2431.
[50]Seckl J R. llbeta-hydroxysteroid dehydrogenases:changing glucocorticoid action[J]. Curr Opin Pharmacol,2004,4(6):597-602.
[51]Lakshmi V, Monder C. Purification and characterization of the corticosteroid 11 beta-dehydrogenase component of the rat liver 11 beta-hydroxysteroid dehydrogenase complex [J]. Endocrinology,1988,123(5): 2390-2398.
[52]Whorwood C B, Mason J I, Ricketts M L, et al. Detection of human 11 beta-hydroxysteroid dehydrogenase isoforms using reverse-transcriptase-polymerase chain reaction and localization of the type 2 isoform to renal collecting ducts[J]. Mol Cell Endocrinol,1995, 110(1-2):R7-R12.
[53]Mairesse J, Lesage J, Breton C, et al. Maternal stress alters endocrine function of the feto-placental unit in rats[J]. Am J Physiol Endocrinol Metab,2007,292(6):E1526-E1533.
[54]Shah P S. Parity and low birth weight and preterm birth:a systematic review and meta-analyses[J]. Acta Obstet Gynecol Scand,2010,89(7): 862-875.
[55]Bertram C, Trowern A R, Copin N, et al. The maternal diet during pregnancy programs altered expression of the glucocorticoid receptor and type 2 llbeta-hydroxysteroid dehydrogenase:potential molecular mechanisms underlying the programming of hypertension in utero[J]. Endocrinology,2001,142(7):2841-2853.
[56]van Beek J P, Guan H, Julan L, et al. Glucocorticoids stimulate the expression of llbeta-hydroxysteroid dehydrogenase type 2 in cultured human placental trophoblast cells[J]. J Clin Endocrinol Metab,2004,89(11): 5614-5621.
[57]Rosewicz S, Mcdonald A R, Maddux B A, et al. Mechanism of glucocorticoid receptor down-regulation by glucocorticoids [J]. J Biol Chem, 1988,263(6):2581-2584.
[58]宋亮年,徐仁宝.糖皮质激素对大鼠糖皮质激素受体mRNA的下降调节[J].第二军医大学学报,1990,(5)：393-397.
[59]Cole T J, Blendy J A, Monaghan A P, et al. Targeted disruption of the glucocorticoid receptor gene blocks adrenergic chromaffin cell development and severely retards lung maturation [J]. Genes Dev,1995,9(13): 1608-1621.
[60]Bhargava A, Fullerton M J, Myles K, et al. The serum- and glucocorticoid-induced kinase is a physiological mediator of aldosterone action[J]. Endocrinology,2001,142(4):1587-1594.
[61]Webster M K, Goya L, Firestone G L. Immediate-early transcriptional regulation and rapid mRNA turnover of a putative serine/threonine protein kinase[J]. J Biol Chem,1993,268(16):11482-11485.
[62]Luca F, Kashyap S, Southard C, et al. Adaptive variation regulates the expression of the human SGK1 gene in response to stress [J]. PLoS Genet, 2009,5(5):e1000489.
[63]Anway M D, Cupp A S, Uzumcu M, et al. Epigenetic transgenerational actions of endocrine disruptors and male fertility[J]. Science,2005, 308(5727):1466-1469.
[64]Harris A, Seckl J. Glucocorticoids, prenatal stress and the programming of disease[J]. Horm Behav,2010.
[65]Whitelaw N C, Whitelaw E. Transgenerational epigenetic inheritance in health and disease[J]. Curr Opin Genet Dev,2008,18(3):273-279.
[66]陈忠,邓卓霖,李坤雄,等.男性不育症睾丸活检诊断的进展[J].广西医科大学学报,1994,(1)：116-119.
[67]苏念军,朱伟杰,李菁,等.亚慢性口服氯化镉对成年大鼠睾丸生精功能的影响[J].生殖与避孕,2005,(3).
[68]Petak S M, Nankin H R, Spark R F, et al. American Association of Clinical Endocrinologists Medical Guidelines for clinical practice for the evaluation and treatment of hypogonadism in adult male patients-2002 update[J]. Endocr Pract,2002,8(6):440-456.
[69]苏念军,朱伟杰,李菁,等.亚慢性口服氯化镉对成年大鼠睾丸生精功能的影响[J].生殖与避孕,2005,(3).
[70]Laboratory manual of the WHO for the examination of human semen and sperm-cervical mucus interaction[J]. Ann 1st Super Sanita,2001,37(1): 1-123.
[71]孙玉成,张建芳,孟令君,等.影响小鼠超数排卵效果的因素[J].北京农学院学报,2005,(4)：17-19.
[72]高建明,侯文元,焦占海.小鼠超数排卵效果分析[J].北京农学院学报,2000,(1)：25-28.
[73]Reik W, Walter J. Genomic imprinting:parental influence on the genome [J]. Nat Rev Genet,2001,2(1):21-32.
[74]Cupp A S, Uzumcu M, Suzuki H, et al. Effect of transient embryonic in vivo exposure to the endocrine disruptor methoxychlor on embryonic and postnatal testis development[J]. J Androl,2003,24(5):736-745.
[75]Uzumcu M, Suzuki H, Skinner M K. Effect of the anti-androgenic endocrine disruptor vinclozolin on embryonic testis cord formation and postnatal testis development and function[J]. Reprod Toxicol,2004,18(6): 765-774.
[76]Carone B R, Fauquier L, Habib N, et al. Paternally induced transgenerational environmental reprogramming of metabolic gene expression in mammals[J]. Cell,2010,143(7):1084-1096.
[77]Rakhshandehroo M, Sanderson L M, Matilainen M, et al. Comprehensive analysis of PPARalpha-dependent regulation of hepatic lipid metabolism by expression profiling[J]. PPAR Res,2007,2007:26839.
[78]Horton T H. Fetal origins of developmental plasticity:animal models of induced life history variation[J]. Am J Hum Biol,2005,17(1):34-43.
[79]Meaney M J, Szyf M, Seckl J R. Epigenetic mechanisms of perinatal programming of hypothalamic-pituitary-adrenal function and health[J]. Trends Mol Med,2007,13(7):269-277.
[80]Weaver I C, Cervoni N, Champagne F A, et al. Epigenetic programming by maternal behavior[J]. Nat Neurosci,2004,7(8):847-854.
[81]Hales C N, Barker D J. The thrifty phenotype hypothesis[J]. Br Med Bull,2001,60:5-20.
[82]Symonds M E, Sebert S P, Hyatt M A, et al. Nutritional programming of the metabolic syndrome[J]. Nat Rev Endocrinol,2009,5(11):604-610.
[83]Bobe P, Chaouat G, Stanislawski M, et al. Immunogenetic studies of spontaneous abortion in mice. II. Antiabortive effects are independent of systemic regulatory mechanisms[J]. Cell Immunol,1986,98(2):477-485.
[84]Clark D A, Chaouat G, Arck P C, et al. Cytokine-dependent abortion in CBA x DBA/2 mice is mediated by the procoagulant fgl2 prothrombinase [correction of prothombinase][J]. J Immunol,1998,160(2):545-549.
[85]Girardi G, Yarilin D, Thurman J M, et al. Complement activation induces dysregulation of angiogenic factors and causes fetal rejection and growth restriction[J]. J Exp Med,2006,203(9):2165-2175.
[86]Seshan S V, Franzke C W, Redecha P, et al. Role of tissue factor in a mouse model of thrombotic microangiopathy induced by antiphospholipid antibodies [J]. Blood,2009,114(8):1675-1683.
[87]郎景和.中国妇产科学的现状与发展[J].国际妇产科学杂志,2008,(1)：3-5.
[88]Henshaw S K. Induced abortion:a world perspective [J]. Perspect Int Planif Fam,1987, (Spec No):12-16.
[89]Lapple M. Stress as an explanatory model for spontaneous abortions and recurrent spontaneous abortions[J]. Zentralbl Gynakol,1988,110(6): 325-335.
[90]Arck P C. Stress and pregnancy loss:role of immune mediators, hormones and neurotransmitters[J]. Am J Reprod Immunol,2001,46(2): 117-123.
[91]Kondoh E, Okamoto T, Higuchi T, et al. Stress affects uterine receptivity through an ovarian-independent pathway. [J]. Hum Reprod,2009, 24(4):945-953.
[92]Nepomnaschy P A, Welch K B, Mcconnell D S, et al. Cortisol levels and very early pregnancy loss in humans [J]. Proc Natl Acad Sci U S A,2006, 103(10):3938-3942.
[93]Zhao Y, Ma R, Shen J, et al. A mouse model of depression induced by repeated corticosterone injections[J]. Eur J Pharmacol,2008,581(1-2): 113-120.
[94]David D J, Samuels B A, Rainer Q, et al. Behavioral effects of fluoxetine in animal madel of anxiety/depression are mediated by both neurogenesis-dependent and independent mechanisms. [J]. Neuron,2009, 62(4):479-493.
[95]Murray F, Smith D W, Hutson P H. Chronic low dose corticosterone exposure decreased hippocampal cell proliferation, volume and induced anxiety and depression like behaviours in mice. [J]. Eur J Pharmacol,2008, 583(1):115-127.
[96]Wang Q, Qian S Y, Yang X L, et al. Effects of different doses of hydrocortisone on the disorder of coagulation in rats at early stage of septic shock[J]. Zhonghua Er Ke Za Zhi,2006,44(1):49-52.
[97]Parker V J, Douglas A J. Stress in early pregnancy:maternal neuro-endocrine-immune responses and effects [J]. J Reprod Immunol,2010, 85(1):86-92.
[98]Rai R, Regan L. Recurrent miscarriage[J]. Lancet,2006,368(9535): 601-611.
[99]Hertig A T, Sheldon W H. Minimal criteria required to prove prima facie case of traumatic abortion or miscarriage:an analysis of 1000 pontaneous abortions. [J]. Ann Surg,1943,117(4):596-606.
[100]刘彩平,张亦心,高章圈,等.211对不孕不育症患者临床诊断分析[J].中国生育健康杂志,2010,(1)：47-48.
[101]倪少凯,黄建初,李晟,等.东莞市不孕不育症患病调查[J].医学与社会, 2010, (8).
[102]Wang H, Dey S K. Roadmap to embryo implantation:clues from mouse models[J]. Nat Rev Genet,2006,7(3):185-199.
[103]郑树民,翟桂霞,韩玉,等.1675例女性不孕不育病因医学鉴定结果分析[J].中国医药指南,2008,(19)：56-58.
[104]Salker M S, Christian M, Steel J H, et al. Deregulation of the serum-and glucocorticoid-inducible kinase SGK1 in the endometrium causes reproductive failure[J]. Nat Med,2011,17(11):1509-1513.
[2]吴尚纯,邱红燕.中国人工流产的现状与对策建议[J].中国医学科学院学报,2010,(5)：479-482.
[3]Shiqian C, Yimin C, Hong W, et al. Study on risk factors of unwanted pregnancy again among the young abortion women in Peking[J]. Maternal and Child Health Care of China,2008, (9):1264-1266.
[4]吴尚纯,邱红燕.中国人工流产的现状与对策建议[J].中国医学科学院学报,2010,(5)：479-482.
[5]郑晓瑛,杨蓉蓉,陈华,等.中国未婚女青年妊娠及流产需要与实现[J].妇女研究论丛,2011,(6)：42-46.
[6]Cheng Y, Gno X, Li Y, et al. Repeat induced abortions and contraceptive practices among unmarried young women seeking an abortion in China[J], International Journal of Gynecology & Obstetrics,2004,87(2):199-202.
[7]Heikinheimo 0, Gissler M, Suhonen S. Age, parity, history of abortion and contraceptive choices affect the risk of repeat abortion[J]. Contraception,2008,78(2):149-154.
[8]Pazol K, Zane S B, Parker W Y, et al. Abortion surveillance--United States,2008[J]. MMWR Surveill Summ,2011,60(15):1-41.
[9]王丽英.人工流产后继发不孕相关因素分析与对策[J].内蒙古中医药,2008,(5)：65-66.
[10]Wu S. Medical abortion in China [J]. J Am Med Womens Assoc,2000,55(3 Suppl):197-199,204.
[11]Creinin M D. Medical abortion regimens:historical context and overview[J]. Am J Obstet Gynecol,2000,183(2 Suppl):S3-S9.
[12]Henshaw S K, Singh S, Haas T. The incidence of abortion worldwide [J]. Int Fam Plann Persp,1999,25(Suppl):S30-S38.
[13]Donaldson K, Briggs J, Mcmaster D. RU 486:an alternative to surgical abortion[J]. J Obstet Gynecol Neonatal Nurs,1994,23(7):555-559.
[14]Westfall J M,O'Brien-Gonzales A, Barley G. Update on early medical and surgical abortion[J]. J Womens Health,1998,7(8):991-995.
[15]张礼婕,刘小平,樊尚荣,等.深圳市人工流产女青少年的性健康状况调查[J].中国全科医学,2004,(15)：1049-1056.
[16]Zhu Q X, Gao E S, Chen A M, et al. Mifepristone-induced abortion and placental complications in subsequent pregnancy[J]. Hum Reprod,2009, 24(2):315-319.
[17]Charles V E, Polis C B, Sridhara S K, et al. Abortion and long-term mental health outcomes:a systematic review of the evidence[J]. Contraception,2008,78(6):436-450.
[18]Gestation, birth-weight, and spontaneous abortion in pregnancy after induced abortion. Report of Collaborative Study by W. H.O. Task Force on Sequelae of Abortion [J]. Lancet,1979,1(8108):142-145.
[19]Atrash H K, Hogue C J. The effect of pregnancy termination on future reproduction[J]. Baillieres Clin Obstet Gynaecol,1990,4(2):391-405.
[20]郑晓瑛,陈功.中国青少年生殖健康可及性调查基础数据报告[J].人口与发展,2010,(3)：2-16.
[21]周亚玲,杨丽华,王秀丽,等.北京市怀柔区妇幼保健院未婚人工流产情况分析[J].中国妇幼保健,2010,(11)：1515-1516.
[22]赵更力,张小松,王临虹,等.中国部分城市流动人口未婚人工流产女青年生殖健康状况分析[J].生殖医学杂志,2005,(5)：16-19.
[23]刘长新.未婚流动人口人工流产相关因素分析[J].中外医疗,2009,(16).
[24]张美萍.青少年生殖健康教育的必要性[J].中国妇幼保健,2009,(23)：3204-3205.
[25]Szekeres-Bartho J, Kinsky R, Chaouat G. The effect of a progesterone-induced immunologic blocking factor on NK-mediated resorption[J]. Am J Reprod Immunol,1990,24(4):105-107.
[26]Szekeres-Bartho J, Par G, Dombay G, et al. The antiabortive effect of progesterone-induced blocking factor in mice is manifested by modulating NK activity[J]. Cell Immunol,1997,177(2):194-199.
[27]Zhang J, Croy B A. Using ultrasonography to define fetal-maternal relationships:moving from humans to mice[J]. Comp Med,2009,59(6): 527-533.
[28]Adamson S L, Lu Y, Whiteley K J, et al. Interactions between trophoblast cells and the maternal and fetal circulation in the mouse placenta[J]. Dev Biol,2002,250(2):358-373.
[29]Lowit A, Bhattacharya S, Bhattacharya S. Obstetric performance following an induced abortion[J]. Best Pract Res Clin Obstet Gynaecol,2010, 24(5):667-682.
[30]Shah P S, Zao J. Induced termination of pregnancy and low birthweight and preterm birth:a systematic review and meta-analyses[J]. BJOG,2009, 116(11):1425-1442.
[31]Sun Y, Che Y, Gao E, et al. Induced abortion and risk of subsequent miscarriage [J]. Int J Epidemiol,2003,32(3):449-454.
[32]Zhou W, Sorensen H T, Olsen J. Induced abortion and low birthweight in the following pregnancy[J]. Int J Epidemiol,2000,29(1):100-106.
[33]Joyce T, Grossman M. The dynamic relationship between low birthweight and induced abortion in New York City. An aggregate time-series analysis [J]. J Health Econ,1990,9(3):273-288.
[34]Shah P S. Parity and low birth weight and preterm birth:a systematic review and meta-analyses[J]. Acta Obstet Gynecol Scand,2010,89(7): 862-875.
[35]Hyatt M A, Budge H, Walker D, et al. Effects of maternal parity and late gestational nutrition on mRNA abundance for growth factors in the liver of postnatal sheep [J]. Am J Physiol Regul Integr Comp Physiol,2007,292(5): R1934-R1942.
[36]Hyatt M A, Keisler D H, Budge H, et al. Maternal parity and its effect on adipose tissue deposition and endocrine sensitivity in the postnatal sheep[J]. J Endocrinol,2010,204(2):173-179.
[37]Lumey L H, Stein A D. Offspring birth weights after maternal intrauterine undernutrition:a comparison within sibships[J]. Am J Epidemiol,1997,146(10):810-819.
[38]Gardner D S, Buttery P J, Daniel Z, et al. Factors affecting birth weight in sheep:maternal environment[J]. Reproduction,2007,133(1): 297-307.
[39]Sun Y, Che Y, Gao E, et al. Induced abortion and risk of subsequent miscarriage [J]. Int J Epidemiol,2003,32(3):449-454.
[40]Zhou W, Olsen J. Are complications after an induced abortion associated with reproductive failures in a subsequent pregnancy?[J]. Acta Obstet Gynecol Scand,2003,82(2):177-181.
[41]Liang H, Gao E S, Chen A M, et al. Mifepristone-induced abortion and vaginal bleeding in subsequent pregnancy[J]. Contraception,2011,84(6): 609-614.
[42]Ahmed A, Singh J, Khan Y, et al. A New Mouse Model to Explore Therapies for Preeclampsia[J]. PLoS One,2010,5(10):e13663.
[43]Zhang J, Sun R, Wei H, et al. Toll-like receptor 3 agonist enhances IFN-gamma and TNF-alpha production by murine uterine NK cells[J]. Int Immunopharmacol,2007,7(5):588-596.
[44]刘智,常才.胎盘功能不足对胎儿的近期和远期影响[J].现代康复,2000,(7)：1110-1111.
[45]Muller M, Renkawitz R. The glucocorticoid receptor[J]. Biochim Biophys Acta,1991,1088(2):171-182.
[46]Tycko B, Efstratiadis A. Genomic imprinting:piece of cake [J]. Nature, 2002,417(6892):913-914.
[47]Redecha P, van Rooijen N, Torry D, et al. Pravastatin prevents miscarriages in mice:role of tissue factor in placental and fetal injury [J]. Blood,2009,113(17):4101-4109.
[48]Osterud B, Bjorklid E. Sources of tissue factor[J]. Semin Thromb Hemost,2006,32(1):11-23.
[49]Redecha P, Tilley R, Tencati M, et al. Tissue factor:a link between C5a and neutrophil activation in antiphospholipid antibody induced fetal injury [J]. Blood,2007,110(7):2423-2431.
[50]Seckl J R. llbeta-hydroxysteroid dehydrogenases:changing glucocorticoid action[J]. Curr Opin Pharmacol,2004,4(6):597-602.
[51]Lakshmi V, Monder C. Purification and characterization of the corticosteroid 11 beta-dehydrogenase component of the rat liver 11 beta-hydroxysteroid dehydrogenase complex [J]. Endocrinology,1988,123(5): 2390-2398.
[52]Whorwood C B, Mason J I, Ricketts M L, et al. Detection of human 11 beta-hydroxysteroid dehydrogenase isoforms using reverse-transcriptase-polymerase chain reaction and localization of the type 2 isoform to renal collecting ducts[J]. Mol Cell Endocrinol,1995, 110(1-2):R7-R12.
[53]Mairesse J, Lesage J, Breton C, et al. Maternal stress alters endocrine function of the feto-placental unit in rats[J]. Am J Physiol Endocrinol Metab,2007,292(6):E1526-E1533.
[54]Shah P S. Parity and low birth weight and preterm birth:a systematic review and meta-analyses[J]. Acta Obstet Gynecol Scand,2010,89(7): 862-875.
[55]Bertram C, Trowern A R, Copin N, et al. The maternal diet during pregnancy programs altered expression of the glucocorticoid receptor and type 2 llbeta-hydroxysteroid dehydrogenase:potential molecular mechanisms underlying the programming of hypertension in utero[J]. Endocrinology,2001,142(7):2841-2853.
[56]van Beek J P, Guan H, Julan L, et al. Glucocorticoids stimulate the expression of llbeta-hydroxysteroid dehydrogenase type 2 in cultured human placental trophoblast cells[J]. J Clin Endocrinol Metab,2004,89(11): 5614-5621.
[57]Rosewicz S, Mcdonald A R, Maddux B A, et al. Mechanism of glucocorticoid receptor down-regulation by glucocorticoids [J]. J Biol Chem, 1988,263(6):2581-2584.
[58]宋亮年,徐仁宝.糖皮质激素对大鼠糖皮质激素受体mRNA的下降调节[J].第二军医大学学报,1990,(5)：393-397.
[59]Cole T J, Blendy J A, Monaghan A P, et al. Targeted disruption of the glucocorticoid receptor gene blocks adrenergic chromaffin cell development and severely retards lung maturation [J]. Genes Dev,1995,9(13): 1608-1621.
[60]Bhargava A, Fullerton M J, Myles K, et al. The serum- and glucocorticoid-induced kinase is a physiological mediator of aldosterone action[J]. Endocrinology,2001,142(4):1587-1594.
[61]Webster M K, Goya L, Firestone G L. Immediate-early transcriptional regulation and rapid mRNA turnover of a putative serine/threonine protein kinase[J]. J Biol Chem,1993,268(16):11482-11485.
[62]Luca F, Kashyap S, Southard C, et al. Adaptive variation regulates the expression of the human SGK1 gene in response to stress [J]. PLoS Genet, 2009,5(5):e1000489.
[63]Anway M D, Cupp A S, Uzumcu M, et al. Epigenetic transgenerational actions of endocrine disruptors and male fertility[J]. Science,2005, 308(5727):1466-1469.
[64]Harris A, Seckl J. Glucocorticoids, prenatal stress and the programming of disease[J]. Horm Behav,2010.
[65]Whitelaw N C, Whitelaw E. Transgenerational epigenetic inheritance in health and disease[J]. Curr Opin Genet Dev,2008,18(3):273-279.
[66]陈忠,邓卓霖,李坤雄,等.男性不育症睾丸活检诊断的进展[J].广西医科大学学报,1994,(1)：116-119.
[67]苏念军,朱伟杰,李菁,等.亚慢性口服氯化镉对成年大鼠睾丸生精功能的影响[J].生殖与避孕,2005,(3).
[68]Petak S M, Nankin H R, Spark R F, et al. American Association of Clinical Endocrinologists Medical Guidelines for clinical practice for the evaluation and treatment of hypogonadism in adult male patients-2002 update[J]. Endocr Pract,2002,8(6):440-456.
[69]苏念军,朱伟杰,李菁,等.亚慢性口服氯化镉对成年大鼠睾丸生精功能的影响[J].生殖与避孕,2005,(3).
[70]Laboratory manual of the WHO for the examination of human semen and sperm-cervical mucus interaction[J]. Ann 1st Super Sanita,2001,37(1): 1-123.
[71]孙玉成,张建芳,孟令君,等.影响小鼠超数排卵效果的因素[J].北京农学院学报,2005,(4)：17-19.
[72]高建明,侯文元,焦占海.小鼠超数排卵效果分析[J].北京农学院学报,2000,(1)：25-28.
[73]Reik W, Walter J. Genomic imprinting:parental influence on the genome [J]. Nat Rev Genet,2001,2(1):21-32.
[74]Cupp A S, Uzumcu M, Suzuki H, et al. Effect of transient embryonic in vivo exposure to the endocrine disruptor methoxychlor on embryonic and postnatal testis development[J]. J Androl,2003,24(5):736-745.
[75]Uzumcu M, Suzuki H, Skinner M K. Effect of the anti-androgenic endocrine disruptor vinclozolin on embryonic testis cord formation and postnatal testis development and function[J]. Reprod Toxicol,2004,18(6): 765-774.
[76]Carone B R, Fauquier L, Habib N, et al. Paternally induced transgenerational environmental reprogramming of metabolic gene expression in mammals[J]. Cell,2010,143(7):1084-1096.
[77]Rakhshandehroo M, Sanderson L M, Matilainen M, et al. Comprehensive analysis of PPARalpha-dependent regulation of hepatic lipid metabolism by expression profiling[J]. PPAR Res,2007,2007:26839.
[78]Horton T H. Fetal origins of developmental plasticity:animal models of induced life history variation[J]. Am J Hum Biol,2005,17(1):34-43.
[79]Meaney M J, Szyf M, Seckl J R. Epigenetic mechanisms of perinatal programming of hypothalamic-pituitary-adrenal function and health[J]. Trends Mol Med,2007,13(7):269-277.
[80]Weaver I C, Cervoni N, Champagne F A, et al. Epigenetic programming by maternal behavior[J]. Nat Neurosci,2004,7(8):847-854.
[81]Hales C N, Barker D J. The thrifty phenotype hypothesis[J]. Br Med Bull,2001,60:5-20.
[82]Symonds M E, Sebert S P, Hyatt M A, et al. Nutritional programming of the metabolic syndrome[J]. Nat Rev Endocrinol,2009,5(11):604-610.
[83]Bobe P, Chaouat G, Stanislawski M, et al. Immunogenetic studies of spontaneous abortion in mice. II. Antiabortive effects are independent of systemic regulatory mechanisms[J]. Cell Immunol,1986,98(2):477-485.
[84]Clark D A, Chaouat G, Arck P C, et al. Cytokine-dependent abortion in CBA x DBA/2 mice is mediated by the procoagulant fgl2 prothrombinase [correction of prothombinase][J]. J Immunol,1998,160(2):545-549.
[85]Girardi G, Yarilin D, Thurman J M, et al. Complement activation induces dysregulation of angiogenic factors and causes fetal rejection and growth restriction[J]. J Exp Med,2006,203(9):2165-2175.
[86]Seshan S V, Franzke C W, Redecha P, et al. Role of tissue factor in a mouse model of thrombotic microangiopathy induced by antiphospholipid antibodies [J]. Blood,2009,114(8):1675-1683.
[87]郎景和.中国妇产科学的现状与发展[J].国际妇产科学杂志,2008,(1)：3-5.
[88]Henshaw S K. Induced abortion:a world perspective [J]. Perspect Int Planif Fam,1987, (Spec No):12-16.
[89]Lapple M. Stress as an explanatory model for spontaneous abortions and recurrent spontaneous abortions[J]. Zentralbl Gynakol,1988,110(6): 325-335.
[90]Arck P C. Stress and pregnancy loss:role of immune mediators, hormones and neurotransmitters[J]. Am J Reprod Immunol,2001,46(2): 117-123.
[91]Kondoh E, Okamoto T, Higuchi T, et al. Stress affects uterine receptivity through an ovarian-independent pathway. [J]. Hum Reprod,2009, 24(4):945-953.
[92]Nepomnaschy P A, Welch K B, Mcconnell D S, et al. Cortisol levels and very early pregnancy loss in humans [J]. Proc Natl Acad Sci U S A,2006, 103(10):3938-3942.
[93]Zhao Y, Ma R, Shen J, et al. A mouse model of depression induced by repeated corticosterone injections[J]. Eur J Pharmacol,2008,581(1-2): 113-120.
[94]David D J, Samuels B A, Rainer Q, et al. Behavioral effects of fluoxetine in animal madel of anxiety/depression are mediated by both neurogenesis-dependent and independent mechanisms. [J]. Neuron,2009, 62(4):479-493.
[95]Murray F, Smith D W, Hutson P H. Chronic low dose corticosterone exposure decreased hippocampal cell proliferation, volume and induced anxiety and depression like behaviours in mice. [J]. Eur J Pharmacol,2008, 583(1):115-127.
[96]Wang Q, Qian S Y, Yang X L, et al. Effects of different doses of hydrocortisone on the disorder of coagulation in rats at early stage of septic shock[J]. Zhonghua Er Ke Za Zhi,2006,44(1):49-52.
[97]Parker V J, Douglas A J. Stress in early pregnancy:maternal neuro-endocrine-immune responses and effects [J]. J Reprod Immunol,2010, 85(1):86-92.
[98]Rai R, Regan L. Recurrent miscarriage[J]. Lancet,2006,368(9535): 601-611.
[99]Hertig A T, Sheldon W H. Minimal criteria required to prove prima facie case of traumatic abortion or miscarriage:an analysis of 1000 pontaneous abortions. [J]. Ann Surg,1943,117(4):596-606.
[100]刘彩平,张亦心,高章圈,等.211对不孕不育症患者临床诊断分析[J].中国生育健康杂志,2010,(1)：47-48.
[101]倪少凯,黄建初,李晟,等.东莞市不孕不育症患病调查[J].医学与社会, 2010, (8).
[102]Wang H, Dey S K. Roadmap to embryo implantation:clues from mouse models[J]. Nat Rev Genet,2006,7(3):185-199.
[103]郑树民,翟桂霞,韩玉,等.1675例女性不孕不育病因医学鉴定结果分析[J].中国医药指南,2008,(19)：56-58.
[104]Salker M S, Christian M, Steel J H, et al. Deregulation of the serum-and glucocorticoid-inducible kinase SGK1 in the endometrium causes reproductive failure[J]. Nat Med,2011,17(11):1509-1513.