骨碎补总黄酮干预Notch信号通路影响骨重建过程中成血管-成骨耦联

Effects of total flavone of Rhizoma Drynariae on the coupling of angiogenesis and osteogenesis in bone remodeling through notch signaling pathway

背景:骨缺损是骨科临床的难题,骨碎补总黄酮对骨缺损治疗有确切疗效,但是其具体机制尚不明确。目的:基于Notch信号通路探讨骨碎补总黄酮在骨重建过程中对血管生成和骨形成的影响。方法:48只SD大鼠随机分为模型组、中药组、中药+DAPT组、DAPT组,每组12只。构建大鼠股骨缺损模型,分别给予中药骨碎补总黄酮和Notch通路阻滞剂DAPT干预,干预12周后通过X射线片、Micro-CT和苏木精-伊红染色观察骨缺损的成骨情况,免疫组化检测骨组织中血管内皮生长因子、CD31、Hes1、骨形态发生蛋白2、Notch1蛋白的表达。结果与结论:①X射线片、Micro-CT和苏木精-伊红染色显示中药组的成骨效果明显优于其他3组;②免疫组化检测显示中药组骨形态发生蛋白2、血管内皮生长因子、CD31、Hes1、Notch1平均吸光度值较模型组明显增多(P<0.05);中药+DAPT组血管内皮生长因子、Hes1、骨形态发生蛋白2的平均吸光度值大于DAPT组(P<0.05),但CD31和Notch1表达组间差异无显著性意义;③结果表明,骨碎补总黄酮通过激活Notch通路促进骨重建过程中成血管-成骨耦联,从而促进骨修复。

BACKGROUND: Bone defect is a clinical difficulty in orthopedics. Total flavone Rhizoma Drynariae has a definite therapeutic effect on bone defects, but thespecific mechanism is not clear.OBJECTIVE: To explore the effects of the total flavone of Rhizoma Drynariae on angiogenesis and bone formation during bone remodeling.METHODS: Forty-eight Sprague-Dawley rats were divided into four groups, 12 in each group, including model group, traditional Chinese medicine (TCM) group,TCM + DAPT group, and DAPT group. femoral bone defect model was established in each rat and interventions with the total flavone of Rhizoma Drynariae andthe Notch signaling pathway blocker DAPT were given in corresponding groups. Osteogenesis was evaluated by X-ray, Micro-CT, and hematoxylin-eosin stainingat 12 weeks of interventions. The levels of vascular endothelial growth factor, CD31, Hes1, bone morphogenetic protein 2, and Notch1 in bone tissue weredetected by immunohistochemistry.RESULTS AND CONCLUSION: X-ray, Micro-CT and hematoxylin-eosin staining showed that TCM group showed significantly better osteogenesis than the otherthree groups. Immunohistochemistry detection indicated that compared with the model group, the average absorbance values of vascular endothelial growthfactor, CD31, Hes1, bone morphogenetic protein 2, and Notch1 were significantly higher in the TCM group (P < 0.05). Compared with the DAPT group, the averageabsorbance values of vascular endothelial growth factor, Hes1, and bone morphogenetic protein 2 were significantly increased in the TCM+DAPT group, while therewas no significant difference in the expression of CD31 and Notch1 between the two groups. These findings indicate that the total flavone of Rhizoma Drynariaecan promote the coupling of angiogenesis-osteogenesis in bone remodeling by activating the Notch pathway, thereby promoting bone repair.