反复严重新生期低血糖大鼠模型的制备及其脑损伤与预后情况  

Establishment of repetitive and profound hypoglycemia model in neonatal rat and associated brain injury and prognosis

作　　者：刘锟[1] 张桂艳[1] 叶信健[1] 张文娟[1] 马炎旭[1] 胡文弋 高红昌[2] 严志汉[1] 

机构地区：[1]温州医科大学附属第二医院育英儿童医院放射科,325000 [2]温州医科大学药学院

出　　处：《中华围产医学杂志》2014年第2期114-118,共5页Chinese Journal of Perinatal Medicine

基　　金：国家自然科学基金（81171306）;浙江省温州市科技局社会发展科学研究项目（Y20100176）;浙江省大学生科技创新活动计划（新苗人才计划）项目（2013R413029）

摘　　要：目的 研究模拟早产儿反复严重新生期低血糖大鼠模型的制作方法,并观察其脑损伤及预后情况. 方法 24只2日龄Sprague-Dawley大鼠随机分为新生期低血糖组、新生期对照组、青春期随访组和青春期对照组,每组6只.研究组（包括新生期低血糖组及青春期随访组）大鼠在生后第2、4、6天腹腔注射胰岛素40 U/kg诱导严重低血糖（血糖≤1.4 mmol/L）,对照组（包括新生期对照组和青春期对照组）予等量生理盐水.对各组大鼠的海马和枕叶皮层行原位末端转移酶标记技术染色,观察反复严重新生期低血糖后新生期及青春期不同部位脑细胞的损伤情况.采用方差分析和t检验进行统计学处理. 结果 在生后第2、4、6天,研究组新生大鼠在给予胰岛素后2.5 h均达到严重低血糖水平（生后第2天＜1.3 mmol/L,生后第4、6天均＜1.4 mmol/L）.与新生期对照组相比,新生期低血糖组海马CAI、CA3、DG区和枕叶皮层内损伤神经元个数增多[新生期低血糖组分别为（23±5）、（28±5）、（234±26）和（187±17）个,新生期对照组分别为（14±3）、（16±3）、（26±4）和（30±6）个,t值分别为2.97、3.11、4.08和3.52,P值均＜0.05],损伤指数升高（新生期低血糖组分别为0.22±0.04、0.24±0.06、0.83±0.06和0.34±0.08,新生期对照组分别为0.12±0.02、0.13±0.02、0.16±0.03和0.17±0.04,t值分别为3.42、3.68、3.92和2.76,P值均＜0.05）.与青春期对照组相比,青春期随访组上述脑区损伤神经元亦增多[青春期随访组分别为（54±7）、（42±9）、（296±34）和（75±12）个,青春期对照组分别为（19±3）、（16±2）、（31±5）和（20±4）个,t值分别为3.47、3.24、3.80和3.92,P值均＜0.05],损伤指数亦升高（青春期随访组分别为0.86±0.12、0.89±0.14、0.92±0.09和0.54±0.11,青春期对照组分别为0.14±0.03、0.13±0.02、0.17±0.02和0.15±0.04,t值分别为3.05、3 60、3.52和3.33,P值均＜0.Objective To investigate the rat model establishment of repetitive and profound hypoglycemia of neonatal period （ RPHN ） , and understand related brain injury and its prognosis. Methods Twenty-four 2-day-old Sprague-Dawley rats were randomly divided into four groups with six in each： the insulin-treated neonatal rats group （ INS-N ） , control neonatal rats group （ CON-N ） , insulin-treated neonatal rats followed up to adolescent stage group （ INS-A ） and control neonatal rats followed up to adolescent stage group （ CON-A ） . Rats in INS-N group and INS-A group received intraperitoneal injections of insulin （ 40 U/kg ） on postnatal day 2, 4 and 6 to induce severe hypoglycemia （ ≤ 1.4 mmol/L ）. The rats in corresponding control group received the same amount of saline. Terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling assay （ TUNEL ） was applied to study the amounts of injured neurons in occipital cortex and hippocampus of the rats in their neonatal period and adolescent period. The difference between the two groups was compared by t test. Results In INS-N and INS-A group, neonatal rats reached severe hypoglycemia level at 2.5 h after insulin injection （ glucose at day 2〈1.3 mmol/L, at day 4 and day 6〈1.4 mmol/L ） . RPHN was observed to induce neuron injury in hippocampus CA1, CA3, DG, and occipital cortex in the INS-N group （numbers of neuron injury in INS-N group： 23±5, 28±5,234±26 and 187± 17, respectively; in CON-N group： 14±3, 16±3, 26±4 and 30±6 ） （ t=-2.97, 3.11, 4.08 and 3.52, all P〈0.05 ） . Compared with CON-N group, INS-N group had higher injured index in above brain regions （ INS-N group： 0.22±0.04, 0.24±0.06, 0.83±0.06 and 0.34±0.08, respectively; CON-N group： 0.12±0.02, 0.13±0.02, 0.16±0.03 and 0.17±0.04 ） （ t=3.42, 3.68, 3.92 and 2.76, all P〈0.05 ） . When the newborn rats reached their adolescent stage, there were more injured neurons in INS-A group in above brain regions （ INS-A group�

关 键 词：婴儿持续高胰岛素血症性低血糖症 脑疾病 疾病模型 动物 预后 

分 类 号：R722.6[医药卫生—儿科] R-332[医药卫生—临床医学]