Lima P H O, Sinzato Y K, Gelaleti R B, Calderon I M P, Rudge M V C, Damasceno D C
Laboratory of Experimental Research in Gynecology and Obstetrics, Department of Gynecology and Obstetrics, Botucatu Medical School, Univ Estadual Paulista, Botucatu, São Paulo State, Brazil.
Exp Clin Endocrinol Diabetes. 2012 May;120(5):303-7. doi: 10.1055/s-0031-1299766. Epub 2012 Mar 15.
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.
本研究旨在评估患有重度或轻度糖尿病的怀孕Wistar大鼠淋巴细胞样本及其新生仔鼠全血样本中的遗传毒性（DNA损伤水平）。使用了Wistar雌性大鼠（1日龄和90日龄）和雄性大鼠（约90日龄）。实验包括2个实验组（每组n = 8只动物）：1）重度糖尿病大鼠，2）轻度糖尿病大鼠。对于轻度糖尿病诱导，大鼠在出生当天皮下注射链脲佐菌素（STZ）（100 mg/kg体重），纳入成年后血糖在120至300 mg/dL之间的大鼠。对于重度糖尿病诱导，成年大鼠接受40 mg/kg STZ（静脉途径），纳入血糖> 300 mg/dL的大鼠。在妊娠第21天，将大鼠麻醉并安乐死，以采集母鼠和胎鼠的血液样本，通过彗星试验应用Endo III和Fpg来测定氧化DNA损伤。因此，轻度糖尿病大鼠及其后代显示出较高的Fpg敏感位点，反映了高血糖导致的损伤。重度糖尿病大鼠及其后代显示出通过Fpg和Endo III敏感位点检测到的较高氧化DNA损伤，表明与糖尿病相关的全身性影响。对DNA损伤的酶处理证明，糖尿病的母体影响与其新生仔鼠的氧化DNA损伤有关，仅使用无酶的DNA损伤分析无法反映这一点。
