基于BMP-2/SMAD/Runx2通路探究骨化三醇胶囊对大鼠胫骨骨折愈合的影响
Effect of calcitriol capsule on tibial fracture healing in rats based on BMP-2/SMAD/Runx2 pathway
  
DOI:10.3969/j.issn.1006-7108.2022.01.010
中文关键词:  骨化三醇胶囊  胫骨  骨折愈合  骨形态发生蛋白2  SMAD  Runt相关转录因子2
英文关键词:calcitriol capsule  tibia  fracture healing  bone morphogenetic protein 2  SMAD  Runt-related transcription factor 2
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王皓* 凃峰 赵文斌 张麟 武汉市第一医院骨科湖北 武汉 430022 
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中文摘要:
      目的 探讨骨化三醇胶囊(Cal)通过骨形态发生蛋白2(BMP-2)/SMAD/Runt相关转录因子2(Runx2)通路对大鼠胫骨骨折愈合的影响及相关机制。方法 将52只SD大鼠随机分为对照组(Cont)、胫骨骨折组、Cal组(灌胃10 ng/mL Cal,连续4周)、Cal+Noggin(BMP特异抑制剂)组(灌胃10 ng/mL Cal+经皮注射50 ng/mL Noggin,连续4周)。采用X射线、全自动生化分析仪、micro CT、HE染色、Western blot分析治疗效果及可能机制。结果 4周后,胫骨骨折组骨折线明显,Cal组骨折线几乎不可见,断端有大量骨痂,Cal+Noggin组骨折愈合较差。胫骨骨折组骨密度、骨体积分数、ALP、OCN、PINP水平及BMP-2、Runx2、p-SMAD1/5/8/SMAD1/5/8表达高于Cont组,骨小梁分离度低于Cont组(P<0.05)。较于胫骨骨折组,Cal组骨密度、骨体积分数、ALP、OCN、PINP水平及BMP-2、Runx2、p-SMAD1/5/8/SMAD1/5/8表达增高,骨小梁分离度降低(P<0.05)。较于Cal组,Cal+Noggin组骨密度、骨体积分数、ALP、OCN、PINP水平及BMP-2、Runx2、p-SMAD1/5/8/SMAD1/5/8表达降低,骨小梁分离度增加(P<0.05)。结论 Cal可能通过激活BMP-2/SMAD/Runx2通路,改善新生骨痂骨密度及微结构,促进胫骨骨折愈合。
英文摘要:
      Objective To explore the effect of calcitriol capsule (Cal) on tibial fracture healing in rats through bone morphogenetic protein-2 (BMP-2)/SMAD/Runt-related transcription factor 2 (Runx2) signaling pathway. Methods Fifty-two SD rats were randomly divided into control group (Cont), tibial fracture group, Cal group (gavage 10 ng/mL Cal for 4 consecutive weeks), and Cal + Noggin (BMP specific inhibitor) group (gavage 10 ng/mL Cal+percutaneous injection of 50 ng/mL Noggin for 4 consecutive weeks). X-ray, automatic biochemical analyzer, micro CT, HE staining and Western blotting were used to analyze the treatment effect and possible mechanism. Results After 4 weeks of the fractures, the fracture line in the tibial fracture group was obvious. The fracture line in the Cal group was almost invisible and there was a lot of callus at the broken end. The fracture healing in the Cal+Noggin group was poor. Bone mineral density, bone volume fraction, levels of ALP, OCN, PINP and the protein levels of BMP-2, Runx2, and p-SMAD1/5/8/SMAD1/5/8 in the tibial fracture group were higher than those in the Cont group (P < 0.05), while trabecular separation was lower than that in the Cont Group (P < 0.05). Compared to those in the tibial fracture group, bone mineral density, bone volume fraction, levels of ALP, OCN, PINP, and the protein levels of BMP-2, Runx2, and p-SMAD1/5/8/SMAD1/5/8 in the Cal group increased, and trabecular separation decreased (P < 0.05). Compared to those in the Cal group, bone mineral density, bone volume fraction, levels of ALP, OCN, PINP, and the protein levels of BMP-2, Runx2, and p-SMAD1/5/8/SMAD1/5/8 in the Cal + Noggin group decreased, and trabecular separation increased (P<0.05). Conclusion Cal may improve bone mineral density and microstructure of new callus, and promote the healing of tibial fractures by activating BMP-2/SMAD/Runx2 pathway.
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