Yin Shao-Wei, Wang Yuan, Meng Yi-Lin, Liu Cai-Xia
Department of Gynecology and Obstetrics, Shengjing Hospital of China Medical University; Key Laboratory of Maternal-Fetal Medicine of Liaoning Province, Shenyang, Liaoning Province, China.
Department of Anesthesiology, Shengjing Hospital of China Medical University, Shenyang, Liaoning Province, China.
Neural Regen Res. 2020 Nov;15(11):2082-2088. doi: 10.4103/1673-5374.282268.
Mild intrauterine hypoperfusion (MIUH) is a serious pathological event that affects the growth and development of fetuses and offspring. MIUH can lead to growth restriction, low birth weight, neurodevelopmental disorders, and other adverse clinical outcomes. To study the effects of MIUH on learning and memory function in offspring, a model of MIUH was established by placing a coil (length 2.5 mm, diameter 0.24 mm) on the uterine artery and ovarian uterine artery of Sprague-Dawley rats in the second trimester of pregnancy (day 17). Next, 120 mg/kg lithium chloride (the MIUH + Li group) or normal saline (the MIUH group) was injected intraperitoneally into these rats. In addition, 120 mg/kg lithium chloride (the Li group) or normal saline (the SHAM group) was injected intraperitoneally into pregnant rats without coil placement. The Morris water maze was used to detect changes in learning and memory ability in the offspring at 4 weeks after birth. In the MIUH group, the escape latency and journey length before reaching the platform were both increased, and the number of times that the platform was crossed and the activity time in the target quadrant within 90 seconds were both decreased compared with the SHAM group. Immunofluorescence double staining and western blot assays demonstrated that hippocampal nestin and Ki67 (both cell-proliferation-related proteins) expression was significantly downregulated in the MIUH group compared with the SHAM group. Furthermore, western blot assays were conducted to investigate changes in related signaling pathway proteins in the brains of offspring rats, and revealed that glycogen synthase kinase 3β (GSK3β) expression was upregulated and β-catenin expression was downregulated in the MIUH group compared with the SHAM group. In addition, compared with the MIUH group, the expression levels of p-GSK3β and β-catenin were upregulated in the MIUH + Li group. These results suggest that MIUH may affect learning and memory function in rat offspring by regulating the GSK3β signaling pathway. The experimental procedures were approved by Animal Ethics Committee of Shengjing Hospital of China Medical University (approval No. 2018PS07K) in June 2018.
轻度宫内灌注不足（MIUH）是一种严重的病理事件，会影响胎儿和子代的生长发育。MIUH可导致生长受限、低出生体重、神经发育障碍及其他不良临床结局。为研究MIUH对子代学习和记忆功能的影响，在妊娠中期（第17天），通过将一个线圈（长度2.5毫米，直径0.24毫米）置于Sprague-Dawley大鼠的子宫动脉和卵巢子宫动脉上，建立了MIUH模型。接下来，向这些大鼠腹腔注射120毫克/千克氯化锂（MIUH + Li组）或生理盐水（MIUH组）。此外，向未放置线圈的孕鼠腹腔注射120毫克/千克氯化锂（Li组）或生理盐水（假手术组）。采用Morris水迷宫检测子代出生后4周时学习和记忆能力的变化。与假手术组相比，MIUH组到达平台前的逃避潜伏期和路程长度均增加，穿过平台的次数以及90秒内在目标象限的活动时间均减少。免疫荧光双染和蛋白质印迹分析表明，与假手术组相比，MIUH组海马巢蛋白和Ki67（两者均为细胞增殖相关蛋白）的表达显著下调。此外，进行蛋白质印迹分析以研究子代大鼠大脑中相关信号通路蛋白的变化，结果显示，与假手术组相比，MIUH组糖原合酶激酶3β（GSK3β）表达上调，β-连环蛋白表达下调。此外，与MIUH组相比，MIUH + Li组中p-GSK3β和β-连环蛋白的表达水平上调。这些结果表明，MIUH可能通过调节GSK3β信号通路影响大鼠子代的学习和记忆功能。实验程序于2018年6月获得中国医科大学附属盛京医院动物伦理委员会批准（批准号：2018PS07K）。
