Exp Clin Endocrinol Diabetes 2012; 120(05): 303-307
DOI: 10.1055/s-0031-1299766
Article
© J. A. Barth Verlag in Georg Thieme Verlag KG Stuttgart · New York

Genotoxicity Evaluation in Severe or Mild Diabetic Pregnancy in Laboratory Animals

P.H. O. Lima
1   Laboratory of Experimental Research in Gynecology and Obstetrics, Department of Gynecology and Obstetrics, Botucatu Medical School, Univ Estadual Paulista_Unesp, Botucatu, São Paulo State, Brazil
,
Y. K. Sinzato
1   Laboratory of Experimental Research in Gynecology and Obstetrics, Department of Gynecology and Obstetrics, Botucatu Medical School, Univ Estadual Paulista_Unesp, Botucatu, São Paulo State, Brazil
,
R. B. Gelaleti
1   Laboratory of Experimental Research in Gynecology and Obstetrics, Department of Gynecology and Obstetrics, Botucatu Medical School, Univ Estadual Paulista_Unesp, Botucatu, São Paulo State, Brazil
,
I.M. P. Calderon
1   Laboratory of Experimental Research in Gynecology and Obstetrics, Department of Gynecology and Obstetrics, Botucatu Medical School, Univ Estadual Paulista_Unesp, Botucatu, São Paulo State, Brazil
,
M.V. C. Rudge
1   Laboratory of Experimental Research in Gynecology and Obstetrics, Department of Gynecology and Obstetrics, Botucatu Medical School, Univ Estadual Paulista_Unesp, Botucatu, São Paulo State, Brazil
,
D. C. Damasceno
1   Laboratory of Experimental Research in Gynecology and Obstetrics, Department of Gynecology and Obstetrics, Botucatu Medical School, Univ Estadual Paulista_Unesp, Botucatu, São Paulo State, Brazil
› Author Affiliations
Further Information

Publication History

received 22 August 2011
first decision 21 December 2011

accepted 22 December 2011

Publication Date:
15 March 2012 (online)

Abstract

This study aimed to evaluate the genotoxicity (DNA damage levels) in lymphocyte samples from pregnant Wistar rats with severe or mild diabetes and in whole blood samples from their newborns. Wistar female rats (1 and 90 days of age) and male rats (approximately 90 days of age) were used. The experiment consisted of 2 experimental groups (n=8 animals/group): 1) rats with severe diabetes, 2) rats with mild diabetes. For mild diabetes induction, the rats received streptozotocin (STZ) subcutaneously (100 mg/kg body weight) at day of birth, and those showing glycemia from 120 to 300 mg/dL in their adult life were included. For induction of severe diabetes, adult rats received 40 mg/kg STZ (intravenous route), and those showing glycemia > 300 mg/dL were included. At day 21 of pregnancy, the rats were anesthetized and euthanized for removal of maternal and fetal blood samples for determination of the oxidative DNA damage by applying Endo III and Fpg using the comet assay. Thus, the rats with mild diabetes and their offspring showed higher Fpg-sensitive sites, reflecting the damage resulting from hyperglycemia. The rats with severe diabetes and their offspring showed higher oxidative DNA damage detected by Fpg and Endo III-sensitive sites, showing general repercussions related to diabetes. The enzymatic treatment for DNA damage evidenced that the maternal repercussions of diabetes are associated with oxidative DNA damage of their newborn, which was not reflected using only the analysis of DNA damage free of the enzymes.

 
  • References

  • 1 Aerts L, Van Assche FA. Animal evidence for the transgenerational development of diabetes mellitus. Int J Biochem Cell Biol 2006; 38 (5-6) 894-903 Epub 2005 Aug 9. Review
  • 2 Anderson D, Yu T-W, Wright J et al. An examinations of DNA strand breakage in the comet assay and antioxidant capacity in diabetic patients. Mutat Res 1998; 398: 151-161
  • 3 Blasiak J, Arabski M, Krupa R et al. DNA damage and repair in type 2 diabetes mellitus. Mutat Res. 2004; 554 (1-2): 297-304
  • 4 Burlinson B, Tice RR, Speit G et al. Fourth International Workgroup on Genotoxicity testing: results of in vivo Comet assay workgroup. Mutat Res 2007; 627: 31-35
  • 5 Collins AR, Duthie SJ, Dobson VL. Direct enzymatic detection of endogenous base damage in human lymphocyte DNA. Carcinogenesis 14 1993; 1733-1735
  • 6 Collins AR, Raslová K, Somorovská M et al. DNA damage in diabetes: correlations with a clinical marker. Free Rad Biol Med 1998; 25 (03) 373-377
  • 7 Damasceno DC, Volpato GT, Mattos IMP et al. Oxidative stress and diabetes in pregnant rats. Anim Reprod Sci 2002; 72 (3-4) 235-244
  • 8 Damasceno DC, Kiss AC, Sinzato YK et al. Maternal-fetal outcome, lipid profile and oxidative stress of diabetic rats neonatally exposed to streptozotocin. Exp Clin Endocrinol Diabetes Jul 2011; a; 119 (07) 408-413 Epub 2011 Jun 10
  • 9 Damasceno DC, Volpato GT, Sinzato YK et al. Genotoxicity and Fetal Abnormality in Streptozotocin-Induced Diabetic Rats Exposed to Cigarette Smoke Prior to and during Pregnancy. Exp Clin Endocrinol Diabetes 2011; b
  • 10 David-Cordonnier MH, Laval J, O’Neill P. Clustered DNA damage, influence on damage excision by XRS5 nuclear extracts and Escherichia coli Nth and Fpg proteins. J Biol Chem Apr 21; 2000; 275 (16) 11865-11873
  • 11 de Souza MS, Sinzato YK, Lima PH et al. Oxidative stress status and lipid profiles of diabetic pregnant rats exposed to cigarette smoke. Reprod Biomed Online Apr 2010; 20 (4): 547-552 Epub 2010 Jan 11
  • 12 Demple B, Harrison L. Repair of oxidative damage to DNA: enzymology and biology. Annu Rev Biochem 1994; 63: 915-948
  • 13 Dinçer Y, Akçay T, Ilkova H et al. DNA damage and antioxidant defense in peripheral leukocytes of patients with Type I diabetes mellitus. Mutat Res Jun 19 2003; 527 (1-2) 49-55
  • 14 Djordjevic A, Spasic S, Jovanovic-Galovic A et al. Oxidative stress in diabetic pregnancy: SOD, CAT and GSH-Px activity and lipid peroxidation products. J Matern Fetal Neonatal Med Dec 2004; 16 (06) 367-372
  • 15 Giacco F, Brownlee M. Oxidative stress and diabetic complications. Circ Res Oct 29 2010; 107 (09) 1058-1070 Review
  • 16 Gontijo AMMC, Tice R. Teste do cometa para detecção de dano no DNA e reparo em células individualiazadas. In: Ribeiro LC, Salavatori DMF, Marques EK. Mutagênese ambiental. 2003: 247-279
  • 17 Goodarzi MT, Navidi AA, Rezaei M et al. Oxidative damage to DNA and lipids: correlation with protein glycation in patients with type 1 diabetes. J Clin Lab Anal 2010; 24 (02) 72-76
  • 18 Grissa O, Atègbo JM, Yessoufou A et al. Antioxidant status and circulating lipids are altered in human gestational diabetes and macrosomia. Transl Res Sep 2007; 150 (03) 164-171 Epub 2007 May 25
  • 19 Hatahet Z, Kow YW, Purmal AA et al. New substrates for old enzymes. 5-Hydroxy-2'-deoxycytidine and 5-hydroxy-2'-deoxyuridine are substrates for Escherichia coli endonuclease III and formamidopyrimidine DNA N-glycosylase, while 5-hydroxy-2'-deoxyuridine is a substrate for uracil DNA N-glycosylase. J Biol Chem Jul 22 1994; 269 (29) 18814-18820
  • 20 Hinokio Y, Suzuki S, Hirai M et al. Oxidative DNA damage in diabetes mellitus: its association with diabetic complications. Diabetologia Aug 1999; 42 (08) 995-998
  • 21 Holemans K, Aerts L, Van Assche FA. Fetal growth restriction and consequences for the offspring in animal models. J Soc Gynecol Investig Oct 2003; 10 (07) 392-399 Review
  • 22 Iessi IL, Bueno A, Sinzato YK et al. Evaluation of neonatally-induced mild diabetes in rats: Maternal and fetal repercussions. Diabetol Metab Syndr Jun 8 2010; 2 (01) 37
  • 23 Lima PH, Damasceno DC, Sinzato YK et al. Levels of DNA damage in blood leukocyte samples from non-diabetic and diabetic female rats and their fetuses exposed to air or cigarette smoke. Mutat Res May 31 2008; 653 (1-2) 44-49 Epub 2008 Mar 14
  • 24 Lima PH, Sinzato YK, de Souza Mda S et al. Evaluation of level of DNA damage in blood leukocytes of non-diabetic and diabetic rat exposed to cigarette smoke. Mutat Res Apr 2 2007; 628 (02) 117-122 Epub 2006 Dec 22
  • 25 Lopez-Soldado I, Herrera E. Different diabetogenic response to moderat doses of streptozotocin in pregnant rats, and its long-term consequences in the offspring. Exp Diabesity Res 2003; 4: 107-118
  • 26 Merzouk H, Madane S, Chabani Sari D et al. Time course of changes in serum glucose, insulin, lipids and tissue lipase activities in macrosomic offspring of rats with streptozotocin-induced diabetes. Clin Sci (London) 2000; 98: 21-30
  • 27 Nishikawa T, Sasahara T, Kiritoshi S et al. Evaluation of urinary 8-hydroxydeoxy-guanosine as a novel biomarker of macrovascular complications in type 2 diabetes. Diabetes Care 2003; 26: 1507-1512
  • 28 Pitozzi V, Giovannelli L, Bardini G et al. Oxidative DNA damage in peripheral blood cells in type 2 diabetes mellitus: higher vulnerability of polymorphonuclear leukocytes. Mutat Res Aug 28 2003; 529 (1-2) 129-133
  • 29 Shimoi K, Okitsu A, Green MH et al. Oxidative DNA damage induced by high glucose and its suppression in human umbilical vein endothelial cells. Mutat Res Sep 1 2001; 480–481: 371-378
  • 30 Sinzato YK, Damasceno DC, Laufer-Amorim R et al. Plasma concentrations and placental immunostaining of interleukin-10 and tumor necrosis factor-α as predictors of alterations in the embryo-fetal organism and the placental development of diabetic rats. Braz J Med Biol Res Mar 2011; 44 (03) 206-211 Epub 2011 Feb 4
  • 31 Sinzato YK, Lima PH, Santos CE et al. Association of diabetes and cigarette smoke exposure on the glycemia and liver glycogen of pregnant Wistar rats. Acta Cir Bras Nov-Dec 2008; 23 (06) 481-485
  • 32 Tchou J, Kasai H, Shibutani S et al. 8-Oxoguanine (8-hydroxyguanine) DNA glycosilase and its substrate specificity. Proc Natl Acad Sci U.S.A 1991; 88: 4690-4694
  • 33 Toescu V, Nuttall SL, Martin U et al. Oxidative stress and normal pregnancy. Clin Endocrinol (Oxf) 57 2002; 609-613
  • 34 Viana M, Aruoma OI, Herrera E et al. Oxidative damage in pregnant diabetic rats and their embryos. Free Radic Biol Med Dec 2000; 29 (11) 1115-1121
  • 35 Volpato GT, Damasceno DC, Rudge MV et al. Effect of Bauhinia forficata aqueous extract on the maternal-fetal outcome and oxidative stress biomarkers of streptozotocin-induced diabetic rats. J Ethnopharmacol Feb 28 2008; 116 (01) 131-137 Epub 2007 Nov 19
  • 36 Wright Jr E, Scism-Bacon JL, Glass LC. Oxidative stress in type 2 diabetes: the role of fasting and postprandial glycemia. Int J Clin Pract 2006; 60 (03) 308-314
  • 37 Wu LL, Chiou CC, Chang PY et al. Urinary 8-OHdG: a marker of oxidative stress to DNA and a risk factor for cancer atherosclerosis and diabetics. Clin Chimica Acta 2004; 339: 1-9
  • 38 Zengi A, Ercan G, Caglayan O et al. Increased Oxidative DNA Damage in Lean Normoglycemic Offspring of Type 2 Diabetic Patients. Exp Clin Endocrinol Diabetes 2011; 119: 467-471
  • 39 Zhao Z, Reece EA. Experimental mechanisms of diabetic embryopathy and strategies for developing therapeutic interventions. J Soc Gynecol Investig 2005; 12 (08) 549-557