目的研究健脾补肾方促进骨质疏松性骨折愈合的疗效机制,探讨"脾肾相关"、"先后天之本"中医理论在骨质疏松性骨折防治中的运用。方法建立去卵巢骨质疏松性骨折小鼠模型,随机分为7、14、28d 3个时间点,每个时间点分生理盐水、健脾补肾方2组。于骨折术后第2天给予相应药物灌胃治疗,分别于治疗3个时间点后处死小鼠,取材,进行影像学观察和病理组织学染色。结果 X-ray、Micro-CT结果显示,健脾补肾组7、14、28d均可促进骨痂形成及骨折愈合;Micro-CT定量分析显示,健脾补肾组7、14d骨痂BV/TV、Conn.D、mean/density of TV、mean/density of BV均明显高于生理盐水组(P〈0.05);有限元分析结果提示,健脾补肾组28d骨折部位刚度、弹性模量结果均高于生理盐水组(P〈0.05);HE染色、ABH/OG染色结果显示,健脾补肾方可以促进骨折部位早、中期膜内成骨和软骨内成骨,加速骨折愈合;免疫组化染色结果显示,健脾补肾方能增加骨痂部位β-catenin、Runx2的表达,促进骨折愈合。结论健脾补肾方可以促进骨质疏松性骨折的愈合,其机制可能与增加β-catenin、Runx2的表达相关。
Objective To study the curative mechanism of the invigorating spleen and nourishing kidney formula on promoting osteoporotic fracture healing,and to discuss the traditional Chinese medicine theories of the correlation between spleen and kidney and the origin of the innateness and acquirement in the application of the osteoporotic fracture prevention. Methods The osteoporotic fracture model was established in ovariectomized mice. The mice were randomly divided into normal saline-treated group and invigorating spleen and nourishing kidney formula-treated group at three time points(7,14,and 28 days). The mice were administered with corresponding drugs by gastric lavage on the second day after the fracture surgery. They were executed at the three time points after the treatment. Samples were collected for image observation and histopathological staining. Results The result of X-ray and micro-CT showed that the formation of callus and fracture healing were promoted in the invigorating spleen and nourishing kidney formula group at 7,14,and 28 days. Micro-CT analysis showed that BV/TV,Conn. D,mean/density of TV,and mean/density of BV in the invigorating spleen and nourishing kidney formula group were significantly higher than those in normal saline group(P〈0. 05) at 7 and 14 days. The finite element analysis suggested that the stiffness of fracture position and the elasticity modulus in invigorating spleen and nourishing kidney formula group were significantly higher than those in normal saline group at 7 days(P〈0. 05). The results of HE and ABH/OG staining showed that the invigorating spleen and nourishing kidney formula promoted intramembranous and endochondral ossification to accelerate fracture healing. The results of immunohistochemistry showed that the invigorating spleen and nourishing kidney formula increased the expression of β-catenin and Runx2 to promote osteoporotic fracture healing. Conclusion The invigorating spleen and nourishing kidney formula effectively promotes osteoporotic fracture h