大鼠发育过程中视网膜低氧诱导因子1α的变化(英文)

Alteration of hypoxia inducible factor-1 alpha in retina during rat development

背景:研究发现,低氧诱导因子1α不仅与缺氧的生理反应有关,而且还参与了正常的胚胎发育过程。目的:观察大鼠发育过程中视网膜低氧诱导因子1α的变化。设计、时间及地点:随机对照动物实验,于2007-01/09在青岛大学医学院山东省分子病毒重点实验室完成。材料:成年未经产清洁型Wistar大鼠,体质量200～250g。方法:雌鼠和雄鼠1∶1合笼,制成孕鼠。分别于孕12,16,20d时剖腹取胎,获得各阶段的鼠胚。出生后大鼠按日龄分别于1,5,10d和12个月龄麻醉处死后摘除眼球,分离视网膜及制作石蜡切片。主要观察指标:免疫组织化学方法、半定量反转录-聚合酶链反应检测大鼠视网膜在胚胎发育过程中各时间段低氧诱导因子1α蛋白及低氧诱导因子1αmRNA的表达。结果:胚胎期视网膜神经上皮层及色素上皮层均有低氧诱导因子1α阳性表达,生后发育早期视网膜神经上皮层及色素上皮层也可见低氧诱导因子1α阳性表达,以神经节细胞、内网层更明显,随着发育进展,其阳性表达主要位于视网膜节细胞层。大鼠视网膜低氧诱导因子1α蛋白及mRNA的表达为胚胎期最高、生后发育期逐渐下降、成年期最低,3期之间相比,差异有显著性意义(P<0.01)。结论:大鼠发育过程中视网膜低氧诱导因子1α随着发育的进展逐渐降低,并逐渐集中表达于视网膜节细胞层。

BACKGROUND： Hypoxia inducible factor-1α （HIF-1α ） is not only related to physiological reaction of hypoxia, but also takes part in normal embryonic development. OBJECTIVE： To study the alteration of HIF-1α in retina during rat development. DESIGN, TIME AND SETTING： Randomized contrast animal study, which was performed in the Shandong Provincial Molecular Virus Key Laboratory, Medical College of Qingdao University between January and September 2007. MATERIALS： Adult Wistar rats, nulliparity, clean grade, and weighing 200-250 g were used in this study. METHODS： Male and female rats were caged as the ratio of 1：1. Embryos were obtained at 12-day, 16-day, and 20-day pregnancy. Eyeballs were obtained from newborn rats by anesthesia at 1 -day, 5-day, 10-day, and 12-month birth. Retina was separated and made into paraffin section. MAIN OUTCOME MEASURES： Expressions of HIF-1 a protein and HIF-1 a mRNA in retina were measured by immunohistochemistry and semiquantitative reverse transcription-polymerase chain reaction at various time points of embryonic development. RESULTS： HIF-1α positively expressed at stratum neuroepitheliale retinae and purpurogenous membrane in the embryonic phase. Additionally, HIF-I a still positively expressed at stratum neuroepitheliale retinae and purpurogenous membrane, especially in ganglionic cells and inner plexiform layer, in early development. With the gradual development, the positive expression was mainly located at stratum ganglionare retinae. HIF-1α protein and mRNA expressions were the highest in the embryonic phase, lower in the development, and the lowest in the adult period. There were significantly differences among these three phases （P 〈 0.01）. CONCLUSION： HIF-1αdecreases gradually in retina and its expression is mainly located at stratum ganglionare retinae.