补肾活血方对大鼠骨折模型的愈合作用及对BMP-2、VEGF及TGF-β1表达的影响

Effect of Bushen Huoxue Decoction on Healing Function of Rat Fracture Model and Its Influence on Expression of BMP-2,VEGF and TGF-β1

目的观察补肾活血方对大鼠骨折模型的愈合作用及对骨形态发生蛋白-2(BMP-2)、血管内皮生长因子(VEGF)及转化生长因子β1(TGF-β1)表达的影响。方法将60只SPF级雄性SD骨折模型大鼠随机分为模型组、中药组和阳性组,模型组给予0.9%氯化钠注射液灌胃,中药组给予补肾活血方灌胃,阳性组给予伤科接骨片灌胃。分别于2周和4周后比较各组大鼠骨组织HE染色、血液流变学指标、ELISA法检测血清炎症因子表达水平和BMP-2、VEGF及转化生长因子β1表达水平,RT-PCR检测BMP-2、VEGF及转化生长因子β1mRNA表达水平。结果模型组骨组织可见破裂、不完整的软骨、纤维性骨痂、炎性细胞浸润、血管生成增多,少量骨小梁形成,大量成骨细胞和成软骨细胞,干预后中药组和阳性组炎性细胞减少,软骨形态有所恢复,纤维性骨痂、软骨细胞减少。干预后中药组和阳性组血浆黏度、全血黏度高切和红细胞聚集指数明显低于模型组,且第4周,中药组显著低于阳性组(P <0.05)。干预后中药组和阳性组血清IL-6和TNF-α水平明显低于模型组,IL-10水平明显高于模型组,且第4周,中药组显著低于阳性组(P <0.05)。干预后中药组和阳性组血清BMP-2、VEGF和TGF-β1及其骨组织mRNA水平明显高于模型组,中药组明显高于阳性组,第4周中药组和阳性组血清BMP-2和TGF-β1及其骨组织mRNA水平明显低于第2周,血清VEGF及其骨组织mRNA水平明显高于第2周(P <0.05)。结论补肾活血方可促进大鼠骨折模型的愈合,其机制可能与改善血液流变学指标,减轻炎症反应,提高BMP-2、VEGF和TGF-β1水平表达有关。

Objective:To investigate the healing effect of Bushen Huoxue Decoction on rat fracture model and the expression of BMP-2,VEGF and TGF-β1. Methods:60 male SPF male SD fracture model rats were randomly divided into the model group,TCM group and the positive group. The model group was given 0.9% normal saline;TCM group was given Bushen Huoxue Decoction,and the positive group was given Shangke Jiegu Tablets by stomach. HE staining,hemorheology and ELISA were used to compare the expression of serum inflammatory cytokines and BMP-2,VEGF and TFG-β1 in bone tissue at 2 and 4 weeks respectively. RT-PCR was used to detect mRNA expression levels of BMP-2,VEGF and TFG-β1. Results:The fractures of the model group showed rupture,incomplete cartilage,fibrous epiphysis,inflammatory cell infiltration,increased angiogenesis,formation of a small amount of trabecular bone,a large number of osteoblasts and chondroblasts. After intervention in TCM group and the positive group,the inflammatory cells were reduced;the cartilage morphology was restored,and fibrous epiphyses and chondrocytes were reduced;plasma viscosity,whole blood viscosity high cut and erythrocyte aggregation index were significantly lower than the model group,and in the fourth week,TCM group was significantly lower than the positive group(P < 0.05). The levels of IL-6 and TNF-α in TCM group were significantly lower than those in the model group;IL-10 level was significantly higher than that in the model group,and in the fourth week,TCM group was significantly lower than the positive group(P < 0.05). The levels of serum BMP-2,VEGF and TGF-β1 and their bone tissue in TCM group and the positive group were significantly higher than those in the model group. TCM group was significantly higher than the positive group. In the 4 th week,those in TCM group and the positive group were significantly lower than those of the second week,and the mRNA levels of serum VEGF and its bone tissue were significantly higher than those of the second week(P < 0.05). Conclusion:Bushen Huoxue Decoction can promote the healing of rat fracture model,and its mechanism may be related to improving blood rheology,reducing inflammation and increasing the expression of BMP-2,VEGF and TGF-β1.