参芪补肾活血汤改善兔股骨骨折延迟愈合的机制初探

Study on the mechanism of Shenqi Bushen Huoxue Decoction in improving delayed femoral fracture healing in rabbits

关键词:  骨延迟愈合  参芪补肾活血汤  BMP-2  TGF-β1  VEGF

Key words:  delayed union of bone  Shenqi Bushen Huoxue Decoction  BMP-2  TGF-β1  VEGF

摘要:

[目的] 初步探讨参芪补肾活血汤对兔股骨骨折延迟愈合的作用效果。[方法] 采用外科手术截除30只新西兰大白兔左侧股骨中段10mm皮质骨的方法构建股骨愈合延迟模型，将模型动物随机分为模型组、参芪补肾活血汤高剂量组（10mL/kg）、参芪补肾活血汤低剂量组（5mL/kg），每组10只，另取10只健康动物作为空白组。参芪补肾活血汤组按剂量每天灌胃1次，持续4周，空白组与模型组同法给予生理盐水。动物给药完成后收集血清样本与愈伤处骨组织样本，逆转录实时定量聚合酶链反应（RT-qPCR）法检测血清中骨形态发生蛋白（BMP）-2、BMP-7、转化生长因子-β1（TGF-β1）和血管内皮生长因子（VEGF）基因相对表达水平，苏木精-伊红染色法观察骨组织愈合情况，免疫组织化学法检测愈伤处骨组织中BMP-2、BMP-7、TGF-β1和VEGF蛋白表达水平。[结果] 病理学显示，模型组骨组织愈合缓慢，损伤区域明显，参芪补肾活血汤高剂量组则出现明显骨痂与骨组织形成。与空白组比较，模型组血清与组织中BMP-2、BMP-7、TGF-β1和VEGF表达水平均降低（P<0.01），与模型组比较，参芪补肾活血汤高剂量血清与组织中BMP-2、BMP-7、TGF-β1和VEGF表达水平均升高（P<0.05）。[结论] 参芪补肾活血汤具有促进兔股骨愈合的效果，其机制可能是促进BMP-2、BMP-7、TGF-β1表达，诱导骨缺损区域血管形成。

Abstract:

[Objective] To explore the effect of Shenqi Bushen Huoxue Decoction on delayed femoral union in rabbits. [Methods] A delayed femoral union model was established by surgically removing 10 mm of cortical bone from the left middle femur of New Zealand white rabbits. The model animals were divided into model group,Shenqi Bushen Huoxue Decoction high-dose group (10 mL/kg) and Shenqi Bushen Huoxue Decoction low-dose group(5 mL/kg),with 10 animals in each group. Another 10 healthy animals were selected as control group. Shenqi Bushen Huoxue Decoction was intragastric for 4 weeks,intragastric administration was given once a day. The control group and model group were given normal saline with the same method. Serum samples and bone tissue samples of callus were collected from the sacrificed animals after drug administration. The relative expression levels of BMP-2,BMP-7,TGF-β1 and VEGF in serum were detected by RT-qPCR. The bone tissue healing was observed by hematoxylin-eosin staining. The expression levels of BMP-2, BMP-7,TGF-β1 and VEGF were detected by immunohistochemistry. [Results] Pathological results showed that the bone tissue in the model group healed slowly and the injured area was obvious,while in the Shenqi Bushen Huoxue Decoction high-dose group,there were obvious callus and bone tissue formation. Compared with control group,the expression levels of BMP-2,BMP-7,TGF-β1 and VEGF in serum and tissues of model group were decreased(P<0.01),compared with model group,the expression levels of BMP-2,BMP-7,TGF-β1 and VEGF in serum and tissues of Shenqi Bushen Huoxue Decoction at high dose were increased(P<0.05). [Conclusion] Shenqi Bushen Huoxue Decoction can promote the healing of rabbits femur,and its mechanism may be to promote the expression of BMP-2,BMP-7 and TGF-β1,and induce the formation of blood vessels in the bone defect area.