Download PDF
Eur J Pediatr Surg 2011; 21(06): 362-365
DOI: 10.1055/s-0031-1285872
Original Article
Georg Thieme Verlag KG Stuttgart · New York

Evaluation of Nitric Oxide (NO) and Nitric Oxide Synthases (NOS) in the Amniotic Fluid in an Experimental Gastroschisis Rat Model

F.L. L. Gonçalves
1   Department of Surgery, School of Medical Sciences, University of Campinas, Campinas, SP, Brazil
,
G.F. P. de Souza
2   Institute of Chemistry, University of Campinas, UNICAMP, Campinas, SP, Brazil
,
A. F. Schmidt
3   Division of Pediatric Surgery, Department of Surgery and Anatomy, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil
,
A. C. Regis
1   Department of Surgery, School of Medical Sciences, University of Campinas, Campinas, SP, Brazil
,
M. G. de Oliveira
2   Institute of Chemistry, University of Campinas, UNICAMP, Campinas, SP, Brazil
,
L. Sbragia
3   Division of Pediatric Surgery, Department of Surgery and Anatomy, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil
› Author Affiliations
Further Information

Publication History

received 22 March 2011

accepted after revision 09 July 2011

Publication Date:
29 September 2011 (online)

    Permissions and Reprints

Abstract

Intestinal damage due to gastroschisis (G), an anomaly found with increasing incidence by pedriatic surgeons, is intimately associated with endogenous nitric oxide (NO) production and NO synthase (NOS) expression.

Aim:

Aim of the study was to evaluate NO production and NOS isoforms in the intestine and amniotic fluid (AF) using a rat model of gastroschisis.

Methods:

A gastroschisis rat model was surgically created at 18.5 days of gestation (term=22 days). 3 groups of 12 fetuses each were studied: control (C), sham (S) and (G). Morphometric data of body weight (BW), intestinal weight (IW) and the IW/BW ratio were evaluated and compared. Indirect quantification of NO (nitrite and nitrate – NOx) was analyzed by chemiluminescence, and the expression of the 3 isoforms was analyzed by Western blotting.

Results:

Group G showed an increase in IW and IW/BW compared with groups C and S. IW: G=0.27±0.06, C=0.20±0.02, S=0.20±0.02 (p<0.01); IW/BW: G=4.11±0.57, C=5.21±1.04, S=5.18±1.23 (p<0.05). NO in the G group was lower in the intestine and higher in AF, as opposed to C and S, where it had increased in the intestine and decreased in AF. Intestinal NOx: G=0.85±0.28, C=1.86±0.82, S=1.80±0.69 (p<0.05); NOx in AF: G=161.87±52.11, C=6.99±5.45, S=48.73±13.183 (p<0.001).

Conclusion:

The intestinal inflammation in gastroschisis promotes the release of nitric oxide to the environment (AF). Perhaps NO in the AF may be an inflammatory marker for G.

Key words

gastroschisis - nitric oxide synthases - experimental surgery - amniotic fluid
 

  • References

  • 1 Ambrose SS. The anterior body wall. In: Skandalakis JE, Gray SW. ed. Embryology for Surgeons. The Embryological Basis for the Treatment of Congenital Anomalies. 2nd edn. Philadelphia, PA: WB Saunders; 1972: 414-415
    Google Scholar
  • 2 Santos MM, Tannuri U, Maksoud JG. Alterations of enteric nerve plexus in experimental gastroschisis: Is there a delay in the maturation?. J Pediatr Surg. 2003. 38. 1506-1511
    Google Scholar
  • 3 Morrison JJ, Klein N, Chitty LS et al. Intra-amniotic inflammation in human gastroschisis: possible aetiology of postnatal bowel dysfunction. Br J Obstet Gynecol 1998; 105: 1200-1204
    PubMedGoogle Scholar
  • 4 Sydorak RM, Sbragia L, Nijagal A et al. Hole in the system: gastroschisis. J Pediatr Surg 2002; 37: 1669-1672
    CrossrefPubMedGoogle Scholar
  • 5 Vanni DS, Horstmann B, Benjo AM et al. Nitric oxide: inhibition of platelets and participation in thrombus formation. J Bras Patol Med Lab 2007; 43: 181-189
    CrossrefPubMedGoogle Scholar
  • 6 Thomas DD, Ridnour La, Isenberg JS et al. The chemical biology of nitric oxide: Implications in celular signaling. Free Radic Biol Med 2008; 45: 18-31
    CrossrefPubMedGoogle Scholar
  • 7 Bealer JF, Graf J, Bruch SW et al. Gastroschisis increases small bowel nitric oxide synthase activity. J Pediatr Surg 1996; 31: 1043-1046
    CrossrefPubMedGoogle Scholar
  • 8 Dilsiz A, Gundogan AH, Aktan M et al. Nitric oxide synthase inhibition prevents intestinal damage in gastroschisis: A morphological evaluation in chick embryos. J Pediatr Surg 1999; 34: 1248-1252
    CrossrefPubMedGoogle Scholar
  • 9 Correia-Pinto J, Tavares ML, Baptista MJ et al. A new fetal rat model of gastroschisis: development and early characterization. J Pediatr Surg 2001; 36: 213-216
    CrossrefPubMedGoogle Scholar
  • 10 Bradford MM. A rapid and sensitive method for a quantitation of microgram quantities of proteins utilizing the principle of protein-dye binding. Anal Biochem 1976; 72: 248-254
    CrossrefPubMedGoogle Scholar
  • 11 Bateman RM, Ellis CG, Freeman DJ. Optimization of nitric oxide chemiluminescence operating conditions for measurement of plasma nitrite and nitrate. Clin Chem 2002; 48: 570-573
    PubMedGoogle Scholar
  • 12 Albert A, Sancho MA, Juliá V et al. Intestinal damage in gastroschisis is independent of the size of the abdominal defect. Pediatr Surg Int 2001; 17: 116-119
    CrossrefPubMedGoogle Scholar
  • 13 Albert A, Margarit J, Julia V et al. Morphology and mucosal biochemistry of gastroschisis intestine in urine-free amniotic fluid. J Pediatr Surg 2003; 38: 1217-1220
    CrossrefPubMedGoogle Scholar
  • 14 França WM, Gonçalves A, Moraes SG et al. Esophageal atresia and other visceral anomalies in a modified Adriamycin rat model and their correlations with amniotic fluid volume variations. Pediatr Surg Int 2004; 20: 602-608
    CrossrefPubMedGoogle Scholar
  • 15 Cheung CY, Brace RA. Amniotic fluid volume and composition in mouse pregnancy. J Soc Gynecol Investig 2005; 12: 558-562
    CrossrefPubMedGoogle Scholar
  • 16 Shepard TH, Tanimura T, Park HW. Glucose absorption and utilization by rat embryos. Int J Dev Biol 1997; 41: 307-314
    PubMedGoogle Scholar
  • 17 Wlodek ME, Westcott KT, Ho PW et al. Reduced fetal, placental, and amniotic fluid PTHrP in the growth-restricted spontaneously hypertensive rat. Am J Physiol Regul Integr Comp Physiol 2000; 279: R31-R38
    PubMedGoogle Scholar
  • 18 Dushi G, Lutz N, Hohlfeld J et al. Morphogenesis of the ureterovesical junction: a histologic and microanatomic study in the rat. Urology 2002; 60: 699-706
    CrossrefPubMedGoogle Scholar
  • 19 Dvorak HF. Vascular permeability to plasma, plasma proteins and cells: an update. Curr Opin Hematol 2010; 17: 225-229
    PubMedGoogle Scholar
  • 20 Di Lorenzo A, Fernandez-Hernando C, Cirino G et al. Akt1 is critical for acute inflammation and histamina-mediated vascular leakage. Proc Natl Acad Sci USA 2009; 106: 14552-14557
    CrossrefPubMedGoogle Scholar
  • 21 Abu-Soud HM, Ichimori K, Nakazawa H et al. Regulation of inducible nitric oxide synthase by self-generated NO. Biochemistry 2001; 40: 6876-6881
    CrossrefPubMedGoogle Scholar