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在家兔模型宫内生长受限胎儿超声心动图和多普勒超声脉冲波

Published: June 29, 2013
doi:
10.3791/50392
, , , , , , , ,
1Division Woman and Child, Department Women,University Hospitals Leuven, 2The Ritchie Centre, Monash Institute of Medical Research, Department of Obstetrics and Gynaecology,Monash University, Victoria, Australia, 3Department of Cardiovascular Sciences,Katholieke Universiteit Leuven, 4Fetal and Perinatal Medicine Research Group,Institut d’Investigacions Biomediques August Pi i Sunyer (IDIBAPS), 5Maternal-Fetal Medicine Department, ICGON, Hospital Clínic,Universitat de Barcelona, Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER)

Summary

我们描述检查胎儿心脏功能与现代功能的胎儿超声心动图和使用VisualSonics VEVO 2100 microultrasound的胎儿胎盘多普勒超声在手术引起的胎儿宫内生长受限模型兔子。

Abstract

胎儿宫内发育迟缓（IUGR）导致心功能异常，这是明显的产前由于胎儿胎盘多普勒超声及胎儿超声心动图的进步。越来越多，这些成像方式被雇用临床检查心脏功能，并在子宫内健康评估，从而指导出生决策时机。在这里，我们使用的胎儿宫内发育迟缓，让兔子模型分析心脏功能的临床相关的方式。使用异氟醚诱导麻醉，胎儿宫内发育迟缓手术创造胎龄25天进行剖腹探查，露出双角子宫，然后结扎每个孕囊在一个单一的子宫角子宫胎盘血管供应的40-50％。在兔双角子宫的其他角作为内部控制胎儿。然后，在胎龄30天（足月）恢复后，同一只兔子进行检查胎儿心脏功能。麻醉诱导与吉胆INE和甲苯噻嗪肌肉注射，然后保持连续静脉滴注氯胺酮和甲苯噻嗪对胎儿的心脏功能，以尽量减少医源性影响。重复剖腹探查进行揭露每个孕囊及胎儿心脏功能的一个microultrasound考试（VEVO VisualSonics 2100）进行。胎盘功能不全是由凸起的搏动指数或缺席或逆转的舒张末期脐动脉多普勒波形流可见一斑。静脉导管和大脑中动脉多普勒检查。胎儿超声心动图检查进行记录B模式，M模式和流速波形在横向和心尖意见。离线计算确定标准的M型心脏变量，三尖瓣和二尖瓣环平面收缩漂移，斑点追踪应变分析，修改后的心肌性能指数和血管血流速度波形的利益。这个小动物模型，胎儿宫内发育迟缓，因此能提供在电脑上检查Ø心脏功能与目前的临床实践是一致的，因此可用于转化研究设置。

Introduction

心血管疾病负担，导致胎儿宫内发育迟缓（IUGR）不能被夸大。它是导致死胎，先天性畸形后1胎儿宫内发育迟缓是指对胎儿未能达到其增长潜力，通常是一个导致胎盘功能不全。2在幸存者中，心肌功能障碍明显的心血管健康状况不佳是显而易见的整个生命跨度在婴儿期和童年3,4和后续的高血压，糖尿病6，肥胖在成年生活中发展-从出生的所有累积的心脏危险因素对缺血性心脏疾病过早死亡7

发展动物模型的特点，建立胎儿宫内发育迟缓和胎儿的后续反应减少基板的可用性母胎通信是必要的，如果我们双方更好地了解胎儿宫内发育迟缓有关汽车的病理生理DIAC减值和心脏保护策略，以提高胎儿和出生后的健康发展。在这方面，许多不同的模式，不同的植物物种已被描述。8胎儿宫内发育迟缓通常产妇营养不足引起低蛋白饮食在啮齿类动物中，9手术切除或结扎子宫血流量在啮齿类动物和豚鼠10或闭塞脐动脉的羊。11然而，很明显，没有模型充分概括了人类胎儿宫内发育迟缓。

在当前的方法论研究，我们采用了很好的验证方法选择性子宫胎盘血管中断，不仅产生超声观察胎儿的心血管反应，临床上14，但也允许审讯胎儿心脏功能新颖的超声心动图使用microultrasound技术在兔13-16的VEVO VisualSonics 2100。虽然多普勒超声fetopl保持acental血管的基石，目前的产前监测胎儿宫内发育迟缓胎儿17，超声心动图功能正在越来越多地被用于疾病的病理生理机制提供新的见解，并以评估胎儿健康18因此，在这里我们采取这些进步从临床研究和描述的动物模型，港口不仅成像的复杂性，但也提供了实验平台调查机制的途径和新的疗法。

Protocol

下面的实验动物伦理委员会，天主教鲁汶大学，比利时鲁汶协议批准。我们跟着前面描述的手术过程13，包括一些方法上的变化，尤其是在麻醉过程。 1。诱导胎儿宫内发育迟缓（IUGR） 于孕兔（混合登德尔蒙德和新西兰白）的时间被安置在个别网箱12/12小时光照时间表与水和标准兔饲料自由采食 ，从胎龄（GA）18天的访问。 GA 25天（足月约是31天），兔?…

Representative Results

一个不对称的生长受限制的子宫胎盘血管结扎的胎儿和胎盘从较正常对照图1F中的胎儿和胎盘。不对称增长证实了减少新生儿出生体重和头围增大：腹围比对照组。从胎儿胎盘多普勒研究的代表性结果示于图2。正端控制胎儿舒张期血流显示一个正常的低电阻模式。随着逐步增加阻力见于胎盘在胎儿宫内发育迟缓的胎儿，没有，然后逆转舒张末流是显而易见的。 图3</…

Discussion

我们已经使用了一个验证的方法，手术子宫胎盘血流量减少，产生胎儿宫内发育迟缓13-16兔子，后来检查胎儿心脏功能14描述microultrasound的技术和可用在VEVO VisualSonics 2100心脏功能分析。繁殖能力，人类胎儿宫内发育迟缓胎儿胎儿胎盘多普勒变化中的小动物模型，然后让最近描述的超声心动图检查心脏功能是胎儿心脏研究可能取得进展，在临床相关的方式。

小?…

Offenlegungen

The authors have nothing to disclose.

Acknowledgements

这项工作是支持汉密尔顿 – 费尔利NHMRC奖学金（RH，AL），维多利亚州政府的运营基础设施支援计划（RH，EW）和的玛丽·居里INDUSTRIA学术界的伙伴关系和途径补助，赞助由欧盟委员会（PD ME） 。笔者想感谢博士弗朗西斯安德烈博士Miyague，俄，罗西塔Kinnart女士和伊万先生Laermans的为他们的技术专长，在生产这种视频。

Materials

Name of Reagent/Material Company Catalog Number Comments
Ketamine Ceva Sante Animale http://www.ceva.com/en
Xylazine Ceva Sante Animale http://www.ceva.com/en
Depot Provera Pharmacia Upjohn
Penicillin G Kela Pharma http://www.kela.be
Lidocaine B Braun Medical http://www.bbraun.com/
Temgesic Schering-Plough http://www.merck-animal-health-usa.com/
Isolurane Isoba Vet; Abbott Laboratories Ltd http://www.abbottanimalhealth.com/index.html
Ethicon Johnson and Johnson http://www.ethiconproducts.co.uk/products/sutures
Ethicon Johnson and Johnson http://www.ethiconproducts.co.uk/products/sutures
Ethicon Johnson and Johnson http://www.ethiconproducts.co.uk/products/sutures
VEVO 2100 VisualSonics SN100-0032 http://www.visualsonics.com/
Aquasonic Gel Parker Laboratories 01 02 http://www.parkerlabs.com/ultrasound_products.html
Nellcor N-20PA Pulse oximeter Covidien http://www.nellcor.com/prod/PRODUCT.ASPX?S1=POX&S2=MON&id=282&V
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Referenzen

  1. Bukowski, R. Stillbirth and fetal growth restriction. Clin. Obstet. Gynecol. 53 (3), 673-680 (2010).
  2. American College of Obstetricians and Gynecologists. Intrauterine growth restriction. ACOG practice bulletin no. 12. 12, (2000).
  3. Crispi, F., Bijnens, B., et al. Fetal growth restriction results in remodeled and less efficient hearts in children. Circulation. 121 (22), 2427-2436 (2010).
  4. Cosmi, E., Visentin, S., Fanelli, T., Mautone, A. J., Zanardo, V. Aortic intima media thickness in fetuses and children with intrauterine growth restriction. Obstet. Gynecol. 114 (5), 1109-1114 (2009).
  5. Ojeda, N. B., Grigore, D., Alexander, B. T. Intrauterine growth restriction: fetal programming of hypertension and kidney disease. Adv. Chronic Kidney Dis. 15 (2), 101-106 (2008).
  6. Stocker, C. J., Arch, J. R., Cawthorne, M. A. Fetal origins of insulin resistance and obesity. Proc. Nutr. Soc. 64 (2), 143-151 (2005).
  7. Barker, D. J. Intrauterine programming of coronary heart disease and stroke. Acta Paediatr. Suppl. 423, 178-182 (1997).
  8. Anthony, R. V., Scheaffer, A. N., Wright, C. D., Regnault, T. R. Ruminant models of prenatal growth restriction. Reprod. Suppl. 61, 183-194 (2003).
  9. Woods, L. L., Weeks, D. A., Rasch, R. Programming of adult blood pressure by maternal protein restriction: role of nephrogenesis. Kidney Int. 65 (4), 1339-1348 (2004).
  10. Turner, A. J., Trudinger, B. J. A modification of the uterine artery restriction technique in the guinea pig fetus produces asymmetrical ultrasound growth. Placenta. 30 (3), 236-2340 (2009).
  11. Miller, S. L., Supramaniam, V. G., Jenkin, G., Walker, D., W, E. M., Wallace, Cardiovascular responses to maternal betamethasone administration in the intrauterine growth-restricted ovine fetus. Am. J. Obstet. Gynecol. 201 (6), 613.e1-613.e8 (2009).
  12. Barry, J. S., Rozance, P. J., Anthony, R. V. An animal model of placental insufficiency-induced intrauterine growth restriction. Semin. Perinatol. 32 (3), 225-2230 (2008).
  13. Eixarch, E., Figueras, F., et al. An experimental model of fetal growth restriction based on selective ligature of uteroplacental vessels in the pregnant rabbit. Fetal Diagn. Ther. 26 (4), 203-211 (2009).
  14. Eixarch, E., Hernandez-Andrade, E., et al. Impact on fetal mortality and cardiovascular Doppler of selective ligature of uteroplacental vessels compared with undernutrition in a rabbit model of intrauterine growth restriction. Placenta. 32 (4), 304-309 (2011).
  15. Eixarch, E., Batalle, D., et al. Neonatal neurobehavior and diffusion MRI changes in brain reorganization due to intrauterine growth restriction in a rabbit model. PLoS One. 7 (2), e31497 (2012).
  16. Figueroa, H., Lozano, M., et al. Intrauterine growth restriction modifies the normal gene expression in kidney from rabbit fetuses. Early Hum. Dev. , (2012).
  17. Alfirevic, Z., Stampalija, T., Gyte, G. M. Fetal and umbilical Doppler ultrasound in high-risk pregnancies. Cochrane Database Syst. Rev. (1), CD007529 (2010).
  18. Baschat, A. A. Examination of the fetal cardiovascular system. Semin. Fetal Neonatal. Med. 16 (1), 2-12 (2011).
  19. Rychik, J., Ayres, N., et al. American Society of Echocardiography guidelines and standards for performance of the fetal echocardiogram. J. Am. Soc. Echocardiogr. 17 (7), 803-810 (2004).
  20. Gnyawali, S. C., Roy, S., Driggs, J., Khanna, S., Ryan, T., Sen, C. K. High-frequency high-resolution echocardiography: first evidence on non-invasive repeated measure of myocardial strain, contractility, and mitral regurgitation in the ischemia-reperfused murine heart. J. Vis. Exp. (41), e1781 (2010).
  21. Forfia, P. R., Fisher, M. R., et al. Tricuspid annular displacement predicts survival in pulmonary hypertension. Am J Respir Crit Care Med. 174 (9), 1034-1041 (2006).
  22. Matos, J., Kronzon, I., Panagopoulos, G., Perk, G. Mitral annular plane systolic excursion as a surrogate for left ventricular ejection fraction. J. Am. Soc. Echocardiogr. 25 (9), 969-974 (2012).
  23. Cruz-Martinez, R., Figueras, F., et al. Normal reference ranges from 11 to 41 weeks’ gestation of fetal left modified myocardial performance index by conventional Doppler with the use of stringent criteria for delimitation of the time periods. Fetal Diagn. Ther. 32 (1-2), 79-86 (2012).
  24. Edwards, A., Baker, L. S., Wallace, E. M. Changes in umbilical artery flow velocity waveforms following maternal administration of betamethasone. Placenta. 24 (1), 12-16 (2003).
  25. Edwards, A., Baker, L. S., Wallace, E. M. Changes in fetoplacental vessel flow velocity waveforms following maternal administration of betamethasone. Ultrasound Obstet. Gynecol. 20 (3), 240-244 (2002).
  26. Neitzke, U., Harder, T., et al. Intrauterine growth restriction in a rodent model and developmental programming of the metabolic syndrome: a critical appraisal of the experimental evidence. Placenta. 29 (3), 246-254 (2008).
  27. Neitzke, U., Harder, T., Plagemann, A. Intrauterine growth restriction and developmental programming of the metabolic syndrome: a critical appraisal. Microcirculation. 18 (4), 304-311 (2011).
  28. Maulik, D., Mundy, D., Heitmann, E. Evidence-based approach to umbilical artery Doppler fetal surveillance in high-risk pregnancies: an update. Clin. Obstet. Gynecol. 53 (4), 869-878 (2010).
  29. Morrow, R. J., Adamson, S. L., Bull, S. B., Ritchie, J. W. Effect of placental embolization on the umbilical arterial velocity waveform in fetal sheep. Am. J. Obstet. Gynecol. 161 (4), 055-60 (1989).
  30. Kingdom, J. C., Burrell, S. J., Kaufmann, P. Pathology and clinical implications of abnormal umbilical artery Doppler waveforms. Ultrasound Obstet. Gynecol. 9 (4), 271-286 (1997).
  31. Van Mieghem, T., DeKoninck, P., Steenhaut, P., Deprest, J. Methods for prenatal assessment of fetal cardiac function. Prenat. Diagn. 29 (13), 1193-1203 (2009).
  32. Van Mieghem, T., Giusca, S., et al. Prospective assessment of fetal cardiac function with speckle tracking in healthy fetuses and recipient fetuses of twin-to-twin transfusion syndrome. J. Am. Soc. Echocardiogr. 23 (3), 301-308 (2010).
  33. Cruz-Martinez, R., Figueras, F., Hernandez-Andrade, E., Oros, D., Gratacos, E. Changes in myocardial performance index and aortic isthmus and ductus venosus Doppler in term, small-for-gestational age fetuses with normal umbilical artery pulsatility index. Ultrasound Obstet. Gynecol. 38 (4), 400-405 (2011).
  34. Comas, M., Crispi, F., Cruz-Martinez, R., Figueras, F., Gratacos, E. Tissue Doppler echocardiographic markers of cardiac dysfunction in small-for-gestational age fetuses. Am. J. Obstet. Gynecol. 205 (1), 57.e1-57.e6 (2011).
  35. Baschat, A. A. Venous Doppler evaluation of the growth-restricted fetus. Clin. Perinatol. 38 (1), 103-112 (2011).
  36. Hodges, R. J., Wallace, E. M. Mending a growth-restricted fetal heart: should we use glucocorticoids?. J. Matern. Fetal Neonatal. Med. , (2012).
  37. Miller, S. L., Chai, M., et al. The effects of maternal betamethasone administration on the intrauterine growth-restricted fetus. Endocrinology. 148 (3), 1288-1295 (2007).
  38. Palahniuk, R. J., Shnider, S. M. Maternal and fetal cardiovascular and acid-base changes during halothane and isoflurane anesthesia in the pregnant ewe. Anesthesiology. 41 (5), 462-472 (1974).
  39. Baumgartner, C., Bollerhey, M., Ebner, J., Laacke-Singer, L., Schuster, T., Erhardt, W. Effects of ketamine-xylazine intravenous bolus injection on cardiovascular function in rabbits. Can. J. Vet. Res. 74 (3), 200-208 (2010).
  40. Cruz-Martinez, R., Figueras, F., et al. Learning curve for Doppler measurement of fetal modified myocardial performance index. Ultrasound Obstet. Gynecol. 37 (2), 158-162 (2011).
  41. Germanakis, I., Gardiner, H. Assessment of fetal myocardial deformation using speckle tracking techniques. Fetal Diagn. Ther. 32 (1-2), 39-46 (2012).
  42. D’hooge, J., Heimdal, A. Regional strain and strain rate measurements by cardiac ultrasound: principles, implementation and limitations. Eur. J. Echocardiogr. 1 (3), 154-170 (2000).
  43. Flake, A. W., Villa, R. L., Adzick, N. S., Harrison, M. R. Transamniotic fetal feeding. II. A model of intrauterine growth retardation using the relationship of “natural runting” to uterine position. J. Pediatr. Surg. 22 (9), 816-819 (1987).

Tags

Fetal EchocardiographyPulsed-wave Doppler UltrasoundRabbit ModelIntrauterine Growth Restriction (IUGR)Placental InsufficiencyCardiac FunctionFetal AnesthesiaDoppler UltrasoundFetal Cardiac ImagingTranslational Research

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Hodges, R., Endo, M., La Gerche, A., Eixarch, E., DeKoninck, P., Ferferieva, V., D’hooge, J., Wallace, E. M., Deprest, J. Fetal Echocardiography and Pulsed-wave Doppler Ultrasound in a Rabbit Model of Intrauterine Growth Restriction. J. Vis. Exp. (76), e50392, doi:10.3791/50392 (2013).
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