摘要
背景:椎间盘终板软骨在一定的循环压力作用下会发生形态学的退变,重要的细胞外基质降低,而同时Wnt-5a基因的表达随着加力时间的延长逐渐降低,Wnt-5a基因与椎间盘退变存在某种重要的内在联系。目的:检测循环压力诱导下Wnt-5a基因在兔椎间盘退变器官模型中的表达变化,探讨其与椎间盘退变之间的关系。方法:选取6月龄新西兰大白兔,取出腰椎间盘制备椎间盘退变器官模型,随机分为加力组和对照组,对照组不加力。利用苏木精-伊红染色及番红O固绿染色观察2组大体组织变化;运用Real time PCR法检测各组软骨中Wnt-5a、Ⅱ型胶原以及蛋白多糖基因的表达,Western-blotting检测Wnt-5a蛋白在各组中表达量的变化。结果与结论:(1)对照组及加力组椎间盘器官培养至第7天,其组织形态学在一定程度上发生改变,大体结构完整,Wnt-5a、Ⅱ型胶原以及蛋白多糖等基因的表达与0 d对照组相比呈现出差异化,14 d加力组以及对照组较0 d组在组织形态学上差异更加明显;(2)7 d加力组、14 d加力组及对照组中的Wnt-5a、Ⅱ型胶原以及蛋白多糖等基因表达下降,Wnt-5a蛋白的表达变化趋势与基因水平检测结果一致,但加力组较未加力各组基因表达降低更加显著,加力组中随加压时间的延长各组基因表达进行性降低;(3)结果提示,通过调节Wnt-5a基因的表达,可能会影响终板软骨细胞的退变进程,进而为椎间盘退变的防治提供新思路。
BACKGROUND:Important extracellular matrixes are reduced with the prolongation of duration of cyclic pressure in the endplate of the intervertebral disc.Meanwhile,the expression of Wnt-5a gene is significantly decreased.There is an important relationship between Wnt-5a gene and intervertebral disc degeneration(IDD).OBJECTIVE:To investigate the expression of Wnt-5a gene under cyclic pressure in a rabbit model of IDD and to explore its role in IDD progress.METHODS:Lumbar intervertebral discs were removed from the 6-month-old New Zealand white rabbits to prepare IDD models and were then randomly divided into experimental(cyclic pressure)and control(no intervention)groups.The morphological changes of intervertebral discs were observed by hematoxylin-eosin staining and safranin O-fast green staining.The mRNA expression levels of proteoglycan,collagen type II,and Wnt-5a were detected by real-time PCR.The protein expression level of Wnt-5a was detected by western blot assay.RESULTS AND CONCLUSION:The morphology of intervertebral discs cultured for 7 days in the experimental and control groups showed a certain change,but was still intact;expression levels of aggrecan,type II collagen,Wnt-5a showed differences from the intervertebral discs cultured for 0 day.On day 14,the damage to the histomorphology was severer in the experimental group than the 0-day control group.The mRNA expression levels of proteoglycan,collagen type II,and Wnt-5a were decreased in both groups,especially the experimental group,at 7 and14 days.The mRNA and protein expression levels of Wnt-5a revealed the same change trend with time.To conclude,regulation of Wnt-5a expression may alter the process of endplate cartilage degeneration,and thus providing new ideas for the prevention and treatment of IDD.
作者
杨晓明
赵泉来
高智
徐宏光
王弘
刘平
Yang Xiao-ming;Zhao Quan-lai;Gao Zhi;Xu Hong-guang;Wang Hong;Liu Ping(Department of Spine Surgery,Yijishan Hospital Affiliated to Wannan Medical University,Wuhu 241001,Anhui Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2018年第4期570-575,共6页
Chinese Journal of Tissue Engineering Research
基金
皖南医学院中青年基金项目(WK2014F02)~~
