GENE THERAPY FOR PULMONARY HYPOPLASIA IN CONGENITAL DIAPHRAGMATIC HERNIA

背景/目的：先天性腹股沟疝（CDH）合并肺发育不全的治疗虽取得了一定进展，但其预后仍较差，胎儿和新生儿期的肺发育被认为是更重要的因素。初步研究表明，肾上腺髓质素（AM）在硝基苯酚处理的大鼠CDH模型中增加分支形态发生。在这项研究中，我们研究了AM基因转移对肺生长的影响，在兔CDH.Methods模型：左侧先天性腹股沟疝手术创建定时怀孕的新西兰白色兔的胎儿在妊娠第24天（任期：31天）。在对照动物中，在妊娠第24天进行剖腹手术。利用带正电的生物可降解明胶制备离子键联的DNA-明胶复合物，可以延缓DNA的降解。结果：在妊娠f30 d，AM明胶组的肺/体重比显著高于明胶组和生理盐水组，但仍显著低于对照组。AM明胶组肺AM较明胶组、生理盐水组和对照组明显增加。免疫组化显示AM明胶组气道细胞和血管细胞中AM染色较强。结论：AM基因转染对兔先天性腹股沟疝有治疗作用。

Background/Purpose : While the prognosis in congenital diaphragmatic hernia (CDH) with hypoplastic lung is still poor in spite of recent progress of the therapy, lung growth in the period of fetus and newborn has been thought to be more important. Preliminary study indicated that adrenomedullin (AM) increased branching morphogenesis in a nitrophen-treated rat model of CDH. In this study, we examined an effect of AM gene transfer on growth of the lung in a rabbit model of CDH.Methods : Left-sided congenital diaphragmatic hernia was surgically created in fetuses of timed-pregnant New Zealand white rabbits on day 24 of gestation (term: 31 days). In control animals, laparotomy was performed on day 24 of gestation. Positively charged biodegradable gelatin was used to produce ionically linked DNA-gelatin complex that could delay DNA degradation. After an incision of the left diaphragm, experimental animals were underwent tracheal injection of AM DNA gelatin complex (AM-gelatin group), gelatin (gelatin group) and saline (saline group).Results : On day f 30 of gestation, lung-to-body weight ratio in AM gelatin group was significantly higher than that in gelatin and saline group, though that was significantly lower than that in controls. Pulmonary AM in AM gelatin group was significantly increased as compared to that in gelatin, saline and control group. Immunohistochemistry in AM gelatin group indicated strong staining of AM in the airway cells and vascular cells. AM m-RNA also increased in AM gelatin group.Conclusion : AM gene transfer induced therapeutic effects in a rabbit model of congenital diaphragmatic hernia.