Maternal-Fetal Outcome, Lipid Profile and Oxidative Stress of Diabetic Rats Neonatally Exposed to Streptozotocin

Abstract

Background: There is no evidence about the integrated issue on glycemia, lipid profile, oxidative stress, and anomaly frequency of pregnant diabetic rats neonatally exposed to streptozotocin.

Objective: Evaluating the impact of hyperglycemia in diabetic rats neonatally exposed to streptozotocin on maternal reproductive and fetal outcomes and the relationship with lipid profile and maternal oxidative stress.

Material and Methods: Ten 90-day-old female Wistar rats were mated to obtain offspring. Some of these newborns received streptozotocin (70 mg/kg, i. p. – n5-STZ group) and the remainder given only citrate buffer (control group) on their day 5 of life. At adult life, these rats (n=13 animals/group) were mated and, at day 21 of pregnancy, they were killed to obtain a maternal blood samples for biochemical determinations. The gravid uterus was weighed with its contents and fetuses were analyzed.

Results: At day 0 of pregnancy, glycemic means of n5-STZ rats were significantly greater compared to those of control rats, but presented fetuses classified as small for pregnancy age. The n5-STZ rats showed increased total cholesterol, triglycerides, MDA concentrations, lower SOD activity and increased frequency fetal visceral anomalies as compared to the control group.

Conclusion: This study showed that the experimental model used led to mild hyperglycemia during pregnancy, although it did not lead to increased macrosomic fetus rates. The hyperglycemic maternal environment caused metabolic alterations, including increased triglyceride and total cholesterol concentrations, and elevated oxidative stress, contributing to increase fetal visceral anomalies.

Key words

mild diabetes - streptozotocin - pregnancy - oxidative stress - anomaly - rat

References

• 1 Aerts L, Van Assche FA. Animal Evidence for the transgenerational development of diabetes mellitus.  Int J Biochem Cell Biol. 2006;  38 894-903
• 2 Bonner-Weir S, Trent DF, Honey RN. et al . Responses of neonatal rat islets to streptozotocin.  Diabetes. 1981;  30 64-69
• 3 Biri A, Bozkurt N, Turp A. et al . Role of oxidative stress in intrauterine growth restriction.  Gynecol Obstet Invest. 2007;  64 187-192
• 4 Calderon IMP, Rudge MVC, Brasil MAM. et al . Diabete e gravidez experimental em ratas I. Indução do diabete, obtenção e evolução da prenhez.  Acta Cir Bras. 1992;  7 142-146
• 5 Caluwaerts S, Holemans K, Van Bree R. et al . Is low-dose Streptozotocin in rats adequate model for gestational Diabetes mellitus?.  J Soc Gynecol Investig. 2003;  10 216-221
• 6 Capobianco E, Jawerbaum A, White V. et al . Elevated levels of endothelin-1 and prostaglandin E2 and their effect on nitric oxide generation in placental tissue from neonatal streptozotocin-induced diabetic rats.  Prostaglandins Leukot Essent Fatty Acids. 2003;  68 225-231
• 7 Damasceno DC, Volpato GT, Calderon IMP. et al . Oxidative stress and diabetes in pregnant rats.  Anim Reprod Sci. 2002;  72 235-244
• 8 Damasceno DC, Volpato GT, Calderon IMP. et al . Effect of Bauhinia forficata extract in diabetic pregnant rats: Maternal repercussions.  Phytomedicine. 2004;  11 196-201
• 9 Damasceno DC, Kempinas WG, Volpato GT. et al .Anomalias Congênitas: Estudos Experimentais.. 1st edn Belo Horizonte: Coopmed; 2008
• 10 Diabetes and Pregnancy Group, France . French multicentric survey of outcome of pregnancy in women with pregestational diabetes.  Diabetes Care. 2003;  26 2990-2993
• 11 Eriksson UJ, Borg LAH. Diabetes and embryonic malformation. Role of substrate-induced free oxygen radical production for dysmorphogenesis in cultured rat embryos.  Diabetes. 1993;  42 411-419
• 12 Eriksson ULF, Dahlaström E, Larsson KS. et al . Increased incidence of congenital malformations in the offspring of diabetic rats and their prevention by maternal insulin therapy.  Diabetes. 1982;  30 1-6
• 13 Eriksson UJ, Cederberg J, Wentzel P. Congenital malformations in offspring of diabetic mothers – animal and human studies.  Rev Endocr Metab Disord. 2003;  4 79-93
• 14 Eriksson UJ, Wenzel P, Hod M. Clinical and experimental advances in the understanding of diabetic embryopathy. In: Hod M, Jovanovic L, Renzo CD, Levia A, Langer O (eds). Textbook of diabetes and pregnancy. 1st edn London: Informa; 2006: 262-275
• 15 Ermis B, Ors R, Yildirim A. et al . Influence of smoking on maternal and neonatal serum malondialdehyde, superoxide dismutase and glutathione peroxidase levels.  Ann Clin Lab Sci. 2004;  34 405-409
• 16 Fatehi-Hassanabad Z, Chan CB, Furman BL. Reactive oxygen species and endothelial function in diabetes.  Eur J Pharmacol. 2010;  636 (1–3) 8-17
• 17 Gelardi NL, Cha CJM, Oh W. Glucose metabolism in adipocytes of obese offspring of mild hyperglycemic rats.  Pediatr Res. 1990;  28 641-645
• 18 Hertogh R, Casi AL, Hinck L. Pre-implantation and embryopathy and maternal diabetes. In: Hod M, Jovanovic L, Renzo CD, Levia A, Langer O, (eds) Textbook of diabetes and pregnancy. 1st edn London: Informa; 2006: 240-252
• 19 Hirano T, Mamo JC, Takeuchi H. et al . Correlation of insulin deficiency and hypertriglyceridemia in diabetic rats.  Diabetes Res Clin Pract. 1991;  12 173-180
• 20 Iessi IL, Bueno A, Sinzato YK. et al . Evaluation of neonatally-induced mild diabetes in rats: Maternal and fetal repercussions.  Diabetol Metab Syndr. 2010;  2 37
• 21 Jansson T, Powell TL. Role of the placenta in fetal programming: underlying mechanisms and potential interventional approaches.  Clin Sci. 2007;  113 1-13
• 22 Karowicz-Bilinska A, Kedziora-Kornatowka K, Bartosz G. Indices of oxidative stress in pregnancy with fetal growth restriction.  Free Radic Res. 2007;  41 870-873
• 23 Kervran A, Guillaume M, Jost A. The endocrine pancreas of the fetus from diabetic pregnant rats.  Diabetologia. 1978;  15 387-393
• 24 Kinney BA, Rabe MB, Jensen RA. et al . Maternal hyperglycemia leads to gender-dependent deficits in learning and memory in offspring.  Exp Biol Med (Maywood). 2003;  228 152-159
• 25 Laudes M, Oberhauser F, Bilkovski R. et al . Human fetal adiponectin and retinol-binding protein (RBP)-4 levels in relation to birth weight and maternal obesity.  Exp Clin Endocrinol Diabetes. 2009;  117 146-149
• 26 Merzouk H, Madane S, Prost J. et al . Changes in serum lipid and lipoprotein concentrations and compositions at birth and after 1 month of life in macrosomic infants of insulin-dependent diabetic mothers.  Eur J Pediatr. 1999;  158 750-756
• 27 Merzouk H, Kahn NA. Implication of lipids in macrosomia of diabetic pregnancy: can n-3 polynsaturated fatty acids exert beneficial effects?.  Clin Sci (Lond). 2003;  105 519-529
• 28 Merzouk H, Madani S, Hichami A. et al . Impaired lipoprotein metabolism in obese offspring streptozotocin-induced rats.  Lipids. 2002;  37 773-781
• 29 Murali B, Goyal RK. Improvement in insulin sensitivity by losartan in non-insulin-dependent diabetic (NIDDM) rats.  Pharmacol Res. 2001;  44 385-389
• 30 Oh W, Gelardi NL, Cha CJM. The cross-generation effect of neonatal macrosomia in rat pups of streptozotocin-induced diabetes.  Pediatr Res. 1991;  29 606-610
• 31 Plagemann A, Harder T, Janert U. et al . Malformations of hypothalamic nuclei in hyperinsulinemic offspring of rats with gestational diabetes.  Dev Neurosci. 1999;  21 58-67
• 32 Portha B, Kergoat M. Dynamics of glucose-induced insulin release during the spontaneous remission of streptozotocin diabetes induced in the newborn rat.  Diabetes. 1985;  34 574-579
• 33 Reece EA, Homko CJ, Wu YK. et al . The role of free radicals and membrane lipids in diabetes-induced congenital malformation.  J Soc Gynecol Investig. 1998;  5 179-198
• 34 Saito FH, Damasceno DC, Kempinas WG. et al . Repercussions of mild diabetes on pregnancy in Wistar rats and on the fetal development.  Diabetol Metab Syndr. 2010;  2 26
• 35 Salewski E. Fabermethode zum markroskopishen nachweis von implantatconsstellen am uterus der ratter naunyn schmuderbergs.  Arch Pharm. 1964;  247 367
• 36 Schaefer-Graf UM, Buchanan TA, Xiang A. et al . Patterns of congenital anomalies and relationship to initial maternal fasting glucose levels in pregnancies complicated by type 2 and gestational diabetes.  Am J Obstet Gynecol. 2000;  182 313-320
• 37 Schwarz KB, Cox JM, Sharma S. et al . Prooxidant effects of maternal smoking and formula in newborn infants.  J Pediatr Gastroenterol Nutr. 1997;  24 68-72
• 38 Shafrir E, Desoye G. Pregnancy in diabetic animals. In: Hod M, Jovanovic L, Renzo CD, Levia A, Langer O (Eds) Textbook of diabetes and pregnancy. 1st edn London: Informa; 2006: 90-112
• 39 Soulimane-Mokhtari NA, Guermouche B, Yessoufou A. et al . Modulation of lipid metabolism by n-3 polyunsaturated fatty acids in gestational diabetic rats and their macrosomic offspring.  Clin Sci (Lond). 2005;  109 287-295
• 40 de Souza MD, Sinzato YK, Lima PH. et al . Oxidative stress status and lipid profiles of diabetic pregnant rats exposed to cigarette smoke.  Reprod Biomed Online. 2010;  20 547-552
• 41 Staples RE, Schnell VL. Refinements in rapid clearing technic in the koh-alizarin red s method for fetal bone.  Stain Technol. 1964;  39 61-63
• 42 Tian F, Luo R, Zhao Z. et al . Blockade of the RAS increases plasma adiponectin in subjects with metabolic syndrome and enhances differentiation and adiponectin expression of human preadipocytes.  Exp Clin Endocrinol Diabetes. 2010;  118 258-265
• 43 Triadou N, Portha B, Picon L. et al . Experimental chemical diabetes and pregnancy in the rat.  Diabetes. 1982;  31 75-79
• 44 Van Assche FA, Holemans K, Aerts L. Fetal growth and consequences for latter life.  J Perinat Med. 1998;  26 337-346
• 45 Vikram A, Tripathi DN, Ramarao P. et al . Evaluation of streptozotocin genotoxicity in rats from different ages using the micronucleus assay.  Regul Toxicol Pharmacol. 2007;  49 238-244
• 46 Volpato GT, Damasceno DC, Kempinas WG. et al . Effect of exercise on the reproductive outcome and fetal development of diabetic rats.  Reprod Biomed Online. 2009;  19 852-858
• 47 Volpato GT, Damasceno DC, Rudge MVC. et al . Effect of Bauhinia forficata aqueous extract on the maternal-fetal outcome and oxidative stress biomarkers of streptozotocin-induced diabetic rats.  J Ethnopharmacol. 2008;  116 131-137
• 48 Watanabe T, Miyagawa I. Caracteristics os ureteral boluts in diabetics rats.  Urol Res. 2002;  30 185-189
• 49 Wilson JG. Methods for administering agents and detecting malformations in experimental animal. In: Wilson JG, Warkany J (eds) Teratology: Principles and Techniques. 1st edn Chicago: University of Chicago Press; 1965: 47-74
• 50 Zangen SW, Yafee P, Kohen R. et al . The role of reactive oxygen species in diabetes-induced anomalies in embryos of Cohen diabetic rats.  Int J Exp Diabetes Res. 2002;  3 247-255
• 51 Zao Z, Reece EA. Experimental mechanisms of diabetic embryopathy and strategies for developing therapeutic interventions.  J Soc Gynecol Investig. 2005;  12 549-557
• 52 Zar JH. Biostatistical Analysis. New Jersey: Prentice-Hall; 1999

Correspondence