Laboratory of Experimental Fetal Surgery "Michael Harrison," Division of Pediatric Surgery, Department of Surgery and Anatomy, School of Medicine of Ribeirao Preto, University of São Paulo Ribeirao Preto, Brazil.
Department of Obstetrics and Gynecology, School of Medical Sciences, University of Campinas, São Paulo, Brazil.
Am J Obstet Gynecol. 2015 Mar;212(3):383.e1-7. doi: 10.1016/j.ajog.2014.09.025. Epub 2014 Sep 28.
To reduce the harmful effect of bowel exposure to amniotic fluid in gastroschisis, we used the nitric oxide (NO) donor S-nitrosoglutathione (GSNO) in an animal model of gastroschisis and assessed the ideal concentration for treatment of changes in bowel.
Gastroschisis was surgically induced in rat fetuses on day 18.5 of gestation. The fetuses were divided into 5 groups (n = 12 animals/group): control (C), gastroschisis (G), gastroschisis + GSNO 5 μmol/L (GNO1), gastroschisis + GSNO 0.5 μmol/L (GNO2), and gastroschisis + GSNO 0.05 μmol/L (GNO3). On day 21.5 of gestation, fetuses were collected by cesarean delivery. Body and intestinal weight were measured and the bowels were either fixed for histometric and immunohistochemical study or frozen for Western blotting. We analyzed bowel morphometry on histological sections and expression of the NO synthase (NOS) enzymes by Western blotting and immunohistochemistry. Data were analyzed by analysis of variance or Kruskal-Wallis test when appropriate.
Morphological and histometric measurements of weight, diameter, and thickness of the layers of the intestinal wall decreased with GSNO treatment, especially in the GNO3 group, when compared with the G group (P < .05). The expression of neuronal NOS, endothelial NOS, and inducible NOS decreased mainly in GNO3 group compared to the G group (P < .05), with no difference compared to C group (P > .05).
Fetal treatment with 0.05 μmol/L GSNO resulted in significant improvement of bowel morphology in gastroschisis.
为了减少先天性腹壁缺损中肠暴露于羊水的有害影响,我们在先天性腹壁缺损动物模型中使用了一氧化氮(NO)供体 S-亚硝基谷胱甘肽(GSNO),并评估了治疗肠改变的理想浓度。
在妊娠第 18.5 天通过手术诱导大鼠胎儿先天性腹壁缺损。胎儿分为 5 组(每组 n = 12 只动物):对照组(C)、先天性腹壁缺损组(G)、先天性腹壁缺损+GSNO5μmol/L 组(GNO1)、先天性腹壁缺损+GSNO0.5μmol/L 组(GNO2)和先天性腹壁缺损+GSNO0.05μmol/L 组(GNO3)。在妊娠第 21.5 天,通过剖宫产收集胎儿。测量体质量和肠质量,将肠固定用于组织学和免疫组织化学研究或冷冻用于 Western blot。我们在组织学切片上分析肠形态计量学,并通过 Western blot 和免疫组织化学分析 NO 合酶(NOS)酶的表达。适当的时候,采用方差分析或 Kruskal-Wallis 检验进行数据分析。
与 G 组相比,GSNO 处理后肠重、直径和肠壁各层厚度的形态学和组织学测量值降低,尤其是 GNO3 组(P <.05)。与 G 组相比,神经元 NOS、内皮 NOS 和诱导型 NOS 的表达主要在 GNO3 组中降低(P <.05),与 C 组相比无差异(P >.05)。
胎儿用 0.05μmol/L GSNO 治疗可显著改善先天性腹壁缺损中的肠形态。