Wnt经典信号通路在兔激素性股骨头坏死中的作用  

The role of Wnt signaling pathway in the femoral head of steroid-induced osteonecrosis in rabbit

作　　者：刘刚[1] 罗高斌[1] 梁晓南[1] 黄捷[1] 李东辉[1] 薄占东[1] Liu Gang Luo Gaobin Liang Xiaonan Huang Jie Li Donghui Bo Zhandong(Department of Bone and Jiont Surgery, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China)

机构地区：[1]广西医科大学第一附属医院骨关节外科,南宁530021

出　　处：《中华实验外科杂志》2016年第11期2472-2476,共5页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（81460348）;广西教育厅一般项目（2013YB314）

摘　　要：目的 观察Wnt经典信号通路相关蛋白和基因在兔激素性股骨头坏死模型中的表达.方法 健康新西兰大白兔72只,雌雄各半,随机分为对照组和模型组各36只.模型组采用内毒素（10 μg/kg）联合甲基强的松龙（20mg/kg）造模,对照组注射等量生理盐水,两组兔于造模后2、4、8周行核磁共振成像（MRI）检查,苏木素-伊红（HE）染色确定模型建立成功;碱性磷酸酶（ALP）染色检测ALP原位表达情况;Western blot和实时荧光定量聚合酶链反应（FQ-PCR）检测Dickkopf相关蛋白-1（DKK-1）、β-连环蛋白（β-catenin）和Runt相关转录因子2（Runx2）表达变化.结果 MRI检查和HE染色结果证实模型建立成功.造模后2、4、8周,模型组兔股骨头内ALP、Runx2和β-catenin表达较对照组降低,而DKK-1较对照组表达增加：模型组ALP和Runx2 mRNA相对表达量为0.73±0.12、0.48±0.12、0.34 ±0.12和0.60±0.10、0.42 ±0.10、0.28±0.11;对照组为1.00 ±0.10、1.00±0.13、1.00±0.10和1.00±0.10、1.00 ±0.11、1.00 ±0.17.模型组ALP和Runx2蛋白表达量为79.80±5.42、68.47±3.90、52.27±4.76和0.58±0.06、0.35±0.04、0.24±0.05;对照组为88.67±6.03、86.67±4.93、93.33±8.50和0.85±0.02、0.81±0.03、0.90±0.07;模型组β-catenin和DKK-1mRNA相对表达量为0.79±0.11、0.44±0.13、0.29±0.10和4.85±0.91、8.08±1.02、12.28±1.51;对照组为1.00±0.07、1.00±0.11、1.00±0.11和1.00±0.10、1.00±0.11、1.00±0.17.模型组β-catenin和DKK-1蛋白表达量为0.18±0.03、0.13±0.03、0.06±0.02和0.12±0.03、0.18±0.02、0.28±0.05;对照组为0.26±0.03、0.25±0.07、0.27±0.05和0.02±0.01、0.02±0.01、0.01 ±0.01,两组比较差异均有统计学意义（P＜0.05）.结论 β-catenin低表达导致成骨分化障碍可能是激素性股骨头坏死发生发展的重要原因.Objective To study the dynamic changes of gene expression and protein synthesis of classical Wnt signaling pathway in the femoral head of steroid-induced osteonecrosis in rabbit.Methods 72 mature rabbits were randomly divided into model group （n =36） and control group （n =36）.Rabbit model of femoral head necrosis was made with lipopolysaccharide plus （LPS,10 μg/kg） methylprednisolone （MPS,20 mg/kg）,the establishment of the model was determined by magnetic resonance imaging （MRI） and hematoxylin-eosin （HE） staining after modeled.Total RNA and protein were extacted from the femoral head.Alkaline phosphatase （ALP） staining,real-time fluorescent quantitative polymerase chain reaction （FQ-PCR） and Western blotting were used to detect the dynamic change of Dickkopf-1 （DKK-1）,ALP,Runt-related transcription factor 2 （Runx2） and β-catenin.Results MRI showed low spot signal on T1 weight and high signal on T2 weight during 2 to 8 weeks,HE staining：bone trabecula became thin and sparse with disorder strcture,abundant apoptoic osteicytees and cells lining cancellous bone were found in model group.Model established successfully.At the post-modeling 2th,4th and 8th week,the expression of ALP,Runx2 and β-catenin gene,protein synthesis in model group were obviously lower than those in control group,while the expression of DKK-1 was extremely higher than those in control group.The volume of ALP and Runx2 mRNA expression were 0.73 ±0.12,0.48 ±0.12,0.34 ± 0.12 and 0.60 ±0.10,0.42 ±0.10,0.28 ±0.11 in model groups,1.00 ±0.10,1.00 ±0.13,1.00 ± 0.10 and 1.00 ±0.10,1.00 ±0.11,1.00 ±0.17 in control groups.The volume of ALP and Runx2 protein expression were 79.80 ± 5.42,68.47 ± 3.90,52.27 ± 4.76 and 0.58 ± 0.06,0.35 ± 0.04,0.24 ± 0.05 in model groups,88.67 ±6.03,86.67 ±4.93,93.33 ±8.50 and 0.85 ± 0.02,0.81 ±0.03,0.90 ± 0.07 in control groups.The volume of β-catenin and DKK-1 mRNA expression were 0.79 ±0.11,0.44±0.13,0.29±0.10 and4.85 ±0.91,8.08 ±1.02,12.28 ±1.51 in

关 键 词：股骨头坏死 WNT信号通路 成骨分化 

分 类 号：R681.8[医药卫生—骨科学]