持续压力对兔纤维环能量代谢相关基因的影响(英文)  

Continuous pressure regulates the expression of energy metabolism related genes in rabbit annulus fibrosus

作　　者：熊晓芊[1] 杨述华[1] 邵增务[1] 詹子睿[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院骨科,湖北省武汉市430022

出　　处：《中国组织工程研究与临床康复》2007年第2期393-396,F0003,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：湖北省自然科学基金资助项目(2004ABA193)~~

摘　　要：背景:近年的体外实验表明,能量代谢相关基因参与了椎间盘细胞对缺氧及营养物质缺乏等不良环境因素的适应机制,但这些基因在压力致椎间盘退变过程中的作用途径尚不十分明确。目的:观察持续压力损伤后,兔纤维环内能量代谢相关基因:低氧诱导因子1α、葡萄糖转运蛋白1和血管内皮生长因子的表达情况。设计:随机对照实验。单位:华中科技大学同济医学院附属协和医院中心实验室及骨科实验室。材料:实验于2005-05/10在华中科技大学同济医学院附属协和医院中心实验室及骨科实验室完成。选用4月龄清洁级日本大白兔25只,体质量2.5～3.0kg。方法:采用可控压力致兔椎间盘退变模型,将术后存活且加压成功的动物随机分为4组:①对照组(n=5):安装加压装置但不予加压。②24h组(n=5):以15kg轴向压力加压24h。③72h组(n=5):以15kg轴向压力加压72h。④恢复组(n=5):以15kg轴向压力加压24h后恢复48h。造模成功麻醉动物处死,取其纤维环标本,应用RT-PCR检测低氧诱导因子1α和葡萄糖转运蛋白1基因的表达,应用WesternBlot及免疫组化检测血管内皮生长因子的含量及分布。主要观测指标:各组低氧诱导因子1α、葡萄糖转运蛋白1基因表达的情况和血管内皮生长因子的含量及分布情况。结果:25只动物完成手术,20只进入结果分析,2只因椎体骨折导致加压失败,3只术后瘫痪并于1～3d内死亡而导致脱失。①低氧诱导因子1α表达情况:对照组仅见少量低氧诱导因子1α表达,24h组低氧诱导因子1α表达较对照组高20余倍(t=25.022,P<0.01),72h组及恢复组低氧诱导因子1α表达较24h组依次下降。②葡萄糖转运蛋白1表达情况:对照组葡萄糖转运蛋白1少量表达,24h组葡萄糖转运蛋白1表达较对照组显著增高(t=18.314,P<0.01),72h组葡萄糖转运蛋白1表达较24h组略增强(t=2.819,P<0.05),恢复组葡萄糖转运蛋白1表达与24h组接近�BACKGROUND： Recent studies have demonstrated that energy metabolism-related genes play important roles in intervertebral disc （IVD） cell adaptation to negative environmental factors, such as hypoxia and insufficiency of nutrient. But the effects of these genes in pressure-induced Intervertebral disc degeneration remain uncertain. OBJECTIVE： To investigate continuous pressure-induced expression of energy metabolism genes： hypoxia-inducible factor 1α（HIF-1α）, glucose transporter-1 （GLUT-1） and vascular endothelial growth factor （VEGF） in rabbit annulus fibrosus （AF）. DESIGN： A randomized controlled experiment SETTING： The Central Laboratory and the Laboratory of Department of Orthopaedics, Union Hospital Affiliated to Tongj Medical College, Huazhong University of Science and Technology. MATERIALS： The experiment was carded out in the Central Laboratory and the Laboratory of Department of Orthopaedics, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology from May to October 2005. Twenty-five Japanese white rabbits（about 4 months old, weighting 2.5-3.0 kg, provided by the Experimental Animal Center of Tongji Medical College, Huazhong University of Science and Technology） were selected to establish the animal model. METHODS ： A controllable pressure-induced rabbit fntervertebral disc degeneration model was adopted to impose vadous pressured on rabbit IVDs in vivo. The survived animals whose IVDs were compressed＇successfully were divided randomly into 4 groups. The IVDs were treated with no pressure as control （control group）, with 15 kg axial load for 24 hours （24 hours group）, 72 hours （72 hours group）, and 24 hours with 48 hours free for self-reparation （reparation group）. Reverse transcriptase-polymerase chain reaction was used to detect the expression of HIF-1α and GLUT-1. Western Blot and immunohistochemical test were carried out for the content and distribution of VEGF. MAIN OUTCOME MEASURES ： Exp

关 键 词：椎间盘 压力 单糖转运蛋白质类 血管内皮生长因子类 疾病模型 动物 

分 类 号：R349.6[医药卫生—基础医学]