Diabetes Mellitus and Teratogenicity - Embryotoxic Effects of Insulin-induced Hypoglycemic Serum during Early Organogenesis in Rat Embryo Culture

我们已经表明，在器官发生的早期阶段暴露于胰岛素诱导的低血糖血清可以不利地影响大鼠胚胎培养物中的胚胎发育，并且这些影响是通过中断糖酵解通量介导的，糖酵解通量构成胚胎该阶段的主要途径（Diabetologia 30：791，1987）。进行进一步的实验以确定在关键时期即使短暂暴露于低血糖血清是否对胚胎发生具有不利影响。孕9.5天的大鼠胚胎在早期头折叠阶段在体外生长48小时，直到神经管关闭。基础培养基由含1.2mg/ml葡萄糖（75%正常大鼠血清）的对照培养基和向对照培养基等渗补充6.0mg/ml葡萄糖的高血糖培养基组成，其中未引起畸形形成性病变。降血糖培养基（0.4mg/ml葡萄糖）是从腹膜内给予胰岛素的大鼠获得的血清。着床后胚胎 ...更多信息 在基础培养期间，在妊娠第10.3天的早期神经管形成中短暂地（1小时）暴露于胰岛素诱导的低血糖血清。在与对照培养基培养期间，短暂暴露于低血糖培养基1小时，胚胎显示轻微的生长迟缓[顶臀（C-R）3.24 <plus-minus>0.05 VS 3.46 <plus-minus>0.05 mm，P&lt;0.01;体节数27.3 <plus-minus>0.4 VS 28.2 <plus-minus>0.1，P&lt;0.05]和小但显著的畸形性病变（7%开放神经孔，13%微脑畸形，11%胚胎畸形）。在添加有低致畸剂量葡萄糖（6.0mg/ml）的高血糖培养基中，低血糖血清暴露1h，生长迟缓（C-R 3.12 <plus-minus>± 0.03 VS 3.35 <plus-minus>± 0.06，P&lt;0.01;体节数26.9 <plus-minus>± 0.02 VS 28.27 <plus-minus>± 0.18，P&lt;0.01）和明显的形态学异常（神经孔开放24%，微脑25%，脑畸形14%，眼畸形10%，心脏畸形5%）。这些结果表明，即使是短暂的低血糖也会对关键发育期的胚胎发生产生不利影响，当胚胎在高血糖环境中培养时，这些影响会增强。

We have shown that exposure to insulin induced-hypoglycemic serum during early phase of organogenesis can adversely affect the embryonic development in rat embryo culture and these effects are mediated through interruption of glycolytic flux that constitutes principal pathway at this stage of embryo (Diabetologia 30: 791, 1987). Further experiments were performed to determine whether or not even brief exposure to hypoglycemic serum during the critical periods has adverse effects on embryogenesis. Rat embryos of 9.5 days of gestation at early head-fold stage were grown in vitro for 48-hr until neural tube closure. Basal culture media consisted of control media containing 1.2mg/ml glucose (75% normal rat serum) and hyperglycemic media supplemented isosmotically with 6.0mg/ml glucose to control media, in which dysmorphogenic lesions were not elicited. Hypoglycemic media (0.4mg/ml glucose) were sera that were obtained from rats given insulin intraperitoneally. Postimplantation embryos were … More briefly (for 1-hr) exposed to the insulin-induced hypoglycemic serum at day 10.3 of gestation in early neural tube formation during the basal culture. On brief exposure to hypoglycemic media for 1-hr during culture with control media, embryos showed minor growth retardation [crown-rump (C-R) 3.24 <plus-minus> 0.05 VS 3.46 <plus-minus> 0.05mm, P<0.01; somite number 27.3 <plus-minus> 0.4 VS 28.2 <plus-minus> 0.1, P<0.05] and small but significant dysmorphogenic lesions (7% open neural pore, 13% microencephaly, 11% skeltal abnormality). Exposure to hypoglycemic serum for 1h during the culture with hyperglycemic media supplemented with subteratogenic dosis of glucose (6.0mg/ml) resulted in greater growth retardation (C-R 3.12 <plus-minus> 0.03 VS 3.35 <plus-minus> 0.06, P<0.01; somite number 26.9 <plus-minus> 0.02 VS 28.27 <plus-minus> 0.18, P<0.01) and marked dysmorphogenic lesions (24% open neural pore, 25% microencepaly, 14% skeltal abnormality, 10% eye abnormality, 5% heart abnormality). These results indicate that even brief hypoglycemia can adversely affect the embryogenesis at critical developing periods and these effects are enhanced when embryogs were cultured in hype Less