摘要
目的观察β-连环蛋白在骨折愈合期间变化,探讨其在骨折愈合期间发挥信号传导作用机制。方法对32只鼠进行随机分组,制作胫骨骨折模型。应用Westernbtot分析对骨折愈合不同时间点β-连环蛋白水平进行测定,逆转录-聚合酶链反应(RT—PCR)技术检测骨折愈合期间Wnts和它们受体表达,组织形态学测量观察β-连环蛋白信号传导拮抗剂Dkk.1对骨折愈合影响。结果β-连环蛋白在骨折愈合期间高表达。Dkk-1腺病毒处理骨痂样本显示β-连环蛋白水平低于未用Dkk1骨痂样本。Wnt-4、Wnt5b以及受体Fz-1和Lrp-6骨折愈合期间呈高表达。Ad—Dkk1组和对照组的骨和软骨占整个骨痂组织量比例、小梁骨直径(μm)、数量(每毫米)、小梁骨间隔(μm)分另0为(11.33±2.52)%/(35.67±3.06)%、(14.33±2.51)%/(47.33±4.04)%、(720.00±77.70)μm/(189.33±83.36)pm、13.67±2.05/32.33±2.62、(1.25±0.96)μm/(4.00±0.82)μm。两组差异有统计学意义(P〈0.01)。结论β-连环蛋白在骨折愈合中发挥重要作用,激活β-连环蛋白的治疗措施可能有助于骨折的愈合。
Objective To observe the changes of β-catenin during fracture repair and explore the mechanism of signaling transduetion through β-eatenin during fracture repair. Methods Mice were randomly divided into three groups, and the animal models of tibia fracture were established. Western blot analysis was employed for detecting the β-catenin expression at different time points during fracture repair. Quantitative RT-PCR was used to detect Wnt ligmants and their receptors during fracture repair to understand whether signaling transduction through β-catenin during fracture healing was directly regulated by Wnts. Histomorphometrie measurements were used to observe the effect of Dkk-1 on fracture repair for further understanding function of β-catenin. Results β-catenin level was low in intact bone tissue,but highly expressed during the entire period of fracture repair. Callus samples from animals treated with Ad-Dkkl exhibited a greater reduction of β-catenin level. Wnt-4, WntSb and their receptors Fz-1 and Lrp-6 were upregulated during fracture healing. In Dkk-1 and eontror groups, using histomorphometric measurements, there was substantial reduction of callus parameters. Bone and cartilage volume as a percentage of total callus tissue volume, trabecular thickness, trabecular number ( per ram) and trabecular separation (OLin) in Dkk-1 and contror groups were ( 11.33± 2.52 ) %/( 35.67 ± 3.06 ) %, ( 14.33 ± 2 51 ) %/( 47.33 ± 4.04) %, (720.00 ± 77.70) μm/( 189.33 ± 83.36) μm, 13.67 ± 2.05/32.33 ± 2.62, ( 1.25 e 0.96 ) μm/( 4.130 ± 0.82) μm respectively ( P 〈 0.01 ). Conclusion β-catenin may play a key signaling role during fracture healing. The therapy to activate β-catenin could be used to enhance fracture repair.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2009年第12期1699-1701,共3页
Chinese Journal of Experimental Surgery