益气健骨补肾法对股骨骨折模型大鼠的作用及机制研究

Therapeutic Effect and Mechanism of Yiqi Jiangu Bushen Method on Femoral Fracture Model Rats

目的:研究益气健骨补肾法对股骨骨折模型大鼠的治疗作用,并初步探讨其作用机制。方法:将60只SD大鼠随机分为空白组、模型组、阿仑膦酸钠组(5g·kg^(-1))及益气健骨补肾方低(2.5g·kg^(-1))、中(5g·kg^(-1))、高(10g·kg^(-1))剂量组,每组10只。除空白组外,其余组大鼠采用闭合骨折法制备股骨骨折模型。造模完成第2天,采用相应药物进行干预,空白组和模型组给予等体积生理盐水,每天1次,连续15d。ELISA法检测大鼠血清Ⅰ型胶原、碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)、转化生长因子-β1(transforming growth factor-β1,TGF-β1)、血管内皮生长因子(vascular endothelial growth factor,VEGF)、抗酒石酸酸性磷酸酶(tartrate resistant acid phosphatase,TRAP)、碱性磷酸酶(alkaline phosphatase,ALP)、Ⅰ型胶原羧基端交联肽(typeⅠcollagen cross-linked C-telopeptide,CTX)及骨钙素(osteocalcin,OC)的水平;双能X线骨密度仪检测大鼠股骨骨密度(bone mineral density,BMD)及组织矿物密度(tissue bone mineral density,TMD);骨组织形态计量仪检测大鼠骨小梁数量(number of trabecular bone,Tb.N)、骨小梁厚度(trabecular thickness,Tb.Th)、骨小梁面积百分数(bone trabecular area percentage,TbAr%)及骨小梁分离度(bone trabecular separation,Tb.Sp);三点弯曲实验检测大鼠股骨最大应力、最大载荷和最大挠度;HE染色观察股骨组织病理变化。结果:与模型组比较,各给药组大鼠血清Ⅰ型胶原、bFGF、TGF-β1、VEGF、ALP、OC的水平及BMD、TMD、Tb.N、Tb.Th、TbAr%、最大应力、最大载荷和最大挠度显著升高(P<0.05),血清TRAP、CTX的水平及Tb.Sp显著降低(P<0.05),股骨组织病理损伤显著改善,且益气健骨补肾方高剂量组的作用效果明显优于阳性对照药阿仑膦酸钠。结论:益气健骨补肾方可显著改善股骨骨折大鼠骨转化指标,增加骨密度,提升骨生物力学性能,其作用机制可能与调控血清中bFGF、TGF-β1、VEGF的水平有关。

Objective:To study the therapeutic effect of Yiqi Jiangu Bushen method on femoral fracture model rats and its mechanism.Methods:Sixty SD rats were randomly divided into blank group,model group,alendronate group(5 mg·kg^(-1))and Yiqi Jiangu Bushen Prescription low(2.5 mg·kg^(-1)),medium(5 mg·kg^(-1))and high(10 mg·kg^(-1))dose groups,with 10 rats in each group.Except the blank group,the femoral fracture model was prepared by closed fracture method in other groups.On the second day after modeling,corresponding drugs were used for intervention.The blank group and model group were given equal volume normal saline once a day for 15 days.ELISA method was used to detect the level of rat serum type I collagen,basic fibroblast growth factor(bFGF),transforming growth factor-β1(TGF-β1),vascular endothelial growth factor(VEGF),tartrate resistant acid phosphatase(TRAP),alkaline phosphatase(ALP),type I collagen cross-linked C-telopeptide(CTX)and osteocalcin(OC).Dual-energy X-ray bone densitometer was used to detect femur bone bone mineral density(BMD)and tissue mineral density(TMD)in rats;the number of trabecular bone(Tb.N),trabecular thickness(Tb.Th),bone trabecular area percentage(TbAr%)and bone trabecular separation(Tb.Sp)in rats;.Three-point bending test was used to detect the maximum stress,maximum load and maximum deflection of rat femur;HE staining was used to observe the pathological changes of femur tissue.Results:Compared with the model group,the serum levels of type I collagen,bFGF,TGF-β1,VEGF,ALP,OC and BMD,TMD,Tb.N,Tb.Th,TbAr%,maximum stress,maximum Load and maximum deflection were significantly increased(P<0.05),serum TRAP,CTX levels and Tb.Sp were significantly reduced(P<0.05),femoral histopathological changes were significantly improved,and the high-dose effect of Yiqi Jiangu Bushen Prescription is significantly better than the positive control drug alendronate sodium.Conclusion:Yiqi Jiangu Bushen Prescription can significantly improve bone transformation indexes,increase bone mineral density and improve bone biomechanical properties in rats with femoral fracture.Its mechanism may be related to the regulation of the level of serum bFGF,TGF-β1 and VEGF.