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妥布霉素激活Wnt/β-catenin信号通路上调ALP和RUNX2蛋白表达促进骨折愈合的实验研究
Hits: 1644   Download times: 473   Received:November 06, 2020    
作者Author单位UnitE-Mail
崔宏刚 CUI Hong-gang 郑州市第七人民医院骨三科, 河南 郑州 450000 The Third Department of Orthopaedics, the 7 th People's Hospital of Zhengzhou, Zhengzhou 450000, Henan, China chg201909@163.com 
期刊信息:《中国骨伤》2021年34卷,第9期,第866-869页
DOI:10.12200/j.issn.1003-0034.2021.09.015


目的:探究妥布霉素(tobramycin,TOB)对大鼠股骨骨折愈合的影响。

方法:选取32只4~6周龄清洁级雄性SD大鼠,体重200~220 g,分为假手术组(A组)、骨折组(B组)、骨折+TOB组(C组)和骨折+TOB+IWR-1组(D组),每组8只。B、C、D组大鼠经闭合性股骨骨折造模法建立骨折模型;A组为假手术组,不做处理;D组大鼠于造模前1 d腹腔注射100 μl (8 μM) Wnt通路抑制剂(IWR-1-endo,IWR-1)。造模1 d后,C、D两组大鼠腹腔注射100 μl (100 μM) TOB,每天1次,共7 d。造模7周后,拍摄X线片观察B、C、D组大鼠的骨折愈合情况;Western-blotting法检测4组大鼠碱性磷酸酶(alkaline phosphatase,ALP),Runt相关转录因子2(Runt-related transcription factor 2,RUNX2)和Wnt信号通路中β-catenin蛋白的表达。

结果:X线片结果显示,C组与造模初期相比,骨折线消失,骨痂形成和骨折愈合良好;B、D两组仍可见少量骨折线,骨痂形成和骨折愈合不良。Western-blotting结果显示,B、C、D组的ALP、RUNX2、β-catenin蛋白表达均高于A组(P<0.05);C组ALP、RUNX2、β-catenin蛋白表达水平较B、D两组升高(P<0.05);应用通路抑制剂IWR-1后,D组ALP、RUNX2、β-catenin蛋白与B组比较差异无统计学意义(P>0.05)。

结论:妥布霉素可以通过激活Wnt/β-catenin信号通路,上调ALP、RUNX2蛋白的表达,促进成骨细胞的分化,进而促进骨折愈合。
[关键词]:妥布霉素  成骨  骨折愈合  Wnt/β-catenin信号通路
 
Tobramycin promotes fracture healing by upregulating expressions of ALP and RUNX2 proteins through activating Wnt/β-catenin pathway
Abstract:

Objective: To explore effect of tobramycin (TOB) on healing of femoral fractures in rats.

Methods: Totally 32 male sprague-dawley (SD) rats were selected and randomly divided into sham group (group A),fracture group (group B),fracture with TOB group (group C) and fracture + TOB + IWR-1 group (group D),8 rats in each group. Close femoral fracture model in rats were established in group B,C and D,group A was sham operation without otherwise process. Group D was intraperitoneal injected 100 μl (8 μM) of Wnt pathway inhibitor IWR-1-endo (IWR-1) before molding at 1 day. At 1 day after molding,100 μl (100 μM) of TOB was intraperitoneally injected into group C and D at once a day for 7 days. At 7 weeks after modling,fracture healing of group B,C and D were observed by X-ray,Western-blotting was appilied to detect alkaline phosphatase(ALP) and Runt-related transcription factor 2 (RUNX2) and β-catenin of Wnt passway.

Results: X-ray results showed fracture line disappeared,callus formation and fracture healing well in group C compared with begning of molding; while a little fracture line,callus formation and fracture malunion in group B and d could be seen. Western-blotting results showed ALP,RUNX2 and expression of β-catenin in group B,C and D were higher than that of group A (P<0.05),while ALP,RUNX2 and β-catenin expression in group C was significantly higher than that of group B and D (P<0.05). After administration of pathway inhibitor IWR-1,there were no significance difference in ALP,RUNX2 and β-catenin protein expression between group D and group B(P>0.05).

Conclusion: Tobramycin could promote osteoblast differentiation and fracture healing by stimulating Wnt/β-catenin signaling pathway,up regulating expression of ALP and RUNX2.
KEYWORDS:Tobramycin  Osteogenesis  Fracture healing  Wnt/β-catenin signaling pathway
 
引用本文,请按以下格式著录参考文献:
中文格式:崔宏刚.妥布霉素激活Wnt/β-catenin信号通路上调ALP和RUNX2蛋白表达促进骨折愈合的实验研究[J].中国骨伤,2021,34(9):866~869
英文格式:CUI Hong-gang.Tobramycin promotes fracture healing by upregulating expressions of ALP and RUNX2 proteins through activating Wnt/β-catenin pathway[J].zhongguo gu shang / China J Orthop Trauma ,2021,34(9):866~869
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