持续激活β-连环蛋白对骨量及骨生长的影响  

Effect of constitutive β-catenin activation on bone mass and bone growth

作　　者：鲍全伟 李昂[1] 刘华渝[1] 李俊峰[1] 沈岳[1] 宗兆文[1] Bao Quanwei Li Ang Liu Huayu Li Junfeng Shen Yue Zong Zhaowen(Department of Trauma, Institute of Surgery Research, Daping Hospital, Third Military Medical University, State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing 400042, Chin)

机构地区：[1]第三军医大学大坪医院野战外科研究所创伤外科创伤、烧伤与复合伤国家重点实验室,重庆400042

出　　处：《中华实验外科杂志》2016年第11期2455-2459,共5页Chinese Journal of Experimental Surgery

基　　金：创伤、烧伤与复合伤国家重点实验室自主课题（SKLZZ201124）

摘　　要：目的 观察在成骨细胞中持续激活β-连环蛋白（β-catenin）对小鼠骨量及骨生长的影响.方法 选取出生后3d基因型为3.2 kb Cre;β-catenin exon 3 FX＋/＋的基因小鼠及同窝野生型小鼠各10只分别作为实验组和对照组,实验组采用腹腔注射他莫昔芬（TM）以持续激活β-catenin（CA-β-catenin）.持续激活1个月后收集胫骨行X线、微焦点X射线断层扫描技术（μCT）检查、苏木素-伊红（HE）染色、组织非特异性碱性磷酸酶染色（TNAP）、Runt相关基因2（Runx2）免疫组织化学染色,抗酒石酸酸性磷酸酶（TRAP）染色,5-溴脱氧尿嘧啶核苷（BrdU）增殖染色,原位缺口末端标记法（TUNEL）法凋亡染色、X-gal染色.结果 CA-β-catenin小鼠与对照组小鼠比较体长变短[（4.1±0.1）cm,P＜0.01],胫骨生长板肥厚但主要表现为肥大层百分比增宽[（77±5）％,P＜0.01]而增殖层变窄[（18 ±3）％,P＜0.01];μCT上表现为CA-β-catenin小鼠皮质骨体积（BV）/组织总体积（TV）[（39±2）％]较对照组小鼠[（9±1）％,P＜0.01]增多;免疫组织化学染色结果显示TNAP阳性细胞数[（22±2）个]及Runx2阳性细胞数增多[（32±4）个,P＜0.01],而TRAP染色阳性结果减少[（3±1）个,P＜0.01];X-gal染色显示3.2 kb-Cre阳性细胞主要分布于松质骨和皮质骨成骨细胞内而生长板软骨内无表达,BrdU染色结果显示增殖层染色阳性细胞数量减少[（9±1）个,P ＜0.01];TUNEL法凋亡染色显示生长板肥大层染色阳性数量减少[（3±1）个,P＜0.01].结论 成骨细胞中持续激活β-catenin导致小鼠骨量增多,但同时出现明显的生长受限.Objective To observe the effect of persistent activation of β-catenin on bone mass and bone growth.Methods A total of 20 mice at 3 days old including 3.2 kb-Cre;β-catenin exon 3 FX ＋/＋ mice and wild-type （as control） mice （n =10） were subjected in this study.X-ray radiology,micro computed tomography （μCT）,hematoxylin and eosin （HE） staining,tartrate resistant acid phosphatase staining （TRAP）,tissue-nonspecific alkaline phosphatase （TNAP）,runt related transcription factor-2 （RUNX2） immunohistochemical staining,5＇-bromo-2＇-deoxyuridine （BrdU） staining and In Situ Cell Death Detection based on TdT-mediated dUTP nick end labeling （TUNEL） technology were employed to observe the effects of CA-β-catenine on bone mass and bone growth after 1 mouth intraperitoneally injected with tamoxifen.Results CA-β-catenine mice was shorter than control mice [（4.1 ±0.1） cm,P 〈 0.01],the growth plates in CA-β-catenine mice [（11.35 ± 0.60） mm] was longer than that in control mice [（7.38 ±0.30） mm,P 〈0.01],and the hypertrophic layer of the growth plate was the most enlarged part [（77 ± 5） %,P 〈 0.01],but the more interesting thing was that the proliferation layer in CA-β-catenine mice [（18 ± 3） %,P 〈 0.01] was shorter than that in control mice [（57 ±6）%,P〈 0.01];μCT examination showed that the bone volume/tissue volume （BV/TV） of CA-β-catenine mice （39 ± 2） % was more than the control mice （9 ± 1） % （P 〈 0.01）.The immunohistochemical staning showed that the number of TNAP （22 ± 2,P 〈 0.01） and Runx2 （32 ± 4,P 〈 0.01）positive cells in CA-β-catenine mice significantly increased when compared with control mice.The TRAP staning showed that the number of osteoclasts in CA-β-catenine mice （3 ± 1,P 〈 0.01） significantly decreased when compared with control mice.The X-gal staning showed that the location of 3.2 kb-Cre positive signals expression only assembled in osteoblasts.The number of BrdU positive cells whi

关 键 词：Β-连环蛋白 成骨细胞 骨量 骨生长 

分 类 号：R735.9[医药卫生—肿瘤]