摘要
目的探讨腺相关病毒介导骨形态发生蛋白-9(BMP-9)对骨质疏松性骨折大鼠的治疗作用。方法60只SD大鼠购自上海维通利华实验动物有限公司,随机分为假手术组、模型组和BMP-9组。模型组和BMP-9组通过摘除双侧卵巢建立骨质疏松大鼠模型,8周后手术折断右侧股骨建立骨质疏松骨折模型。假手术组仅摘除卵巢周围脂肪组织。BMP-9组大鼠于骨折部位注射过表达BMP-9的腺相关病毒1×1010 vg,假手术组和模型组于骨折部位注射对照腺相关病毒1×1010 vg。分别取注射后第7周模型大鼠,X线摄片后用半定量评分法对骨折恢复情况进行评分;测定骨折处行骨密度检测;生物力学检测骨折愈合的力学参数;采用酶联免疫吸附试验(ELISA)检测骨代谢指标变化;采用荧光定量聚合酶链反应(PCR)分析骨折部位血管内皮生长因子(VEGF)和核因子-κB(NF-κB)受体激活物配体(RANKL)的mRNA水平。计量数据比较采用方差分析,组间比较采用LSD法。结果与模型组(1.15±0.21)比较,BMP-9组大鼠骨折部位BMP-9 mRNA水平(3.94±0.34)显著增加,差异有统计学意义(t=3.089,P<0.05)。与模型组X线评分和骨密度[(2.23±0.25)分、0.077±0.002]比较,BMP-9组大鼠X线评分和骨密度显著增加[(4.10±0.31)分、0.023±0.004],差异有统计学意义(t=2.917、3.091,P<0.05)。与模型组碱性磷酸酶(ALP)、骨保护素(OPG)和骨钙素(BGP)水平[(20.33±3.05)U/L、(1.12±0.07)μg/L、(2.21±0.36)μg/L]比较,BMP-9组大鼠血清ALP、OPG和BGP水平[(29.18±3.29)U/L、(1.39±0.09)μg/L、(4.48±0.51)μg/L]显著增加,差异有统计学意义(t=2.099、2.514、2.814,P<0.05)。与模型组比较,BMP-9组大鼠股骨生物力学参数显著改善,差异有统计学意义(t=2.471、2.104、2.810,P<0.05)。与模型组VEGF和RANKL水平(1.23±0.24、1.00±0.19)比较,BMP-9组大鼠VEGF和RANKL水平(2.58±0.31、3.29±0.22)显著增加,差异有统计学意义(t=2.034、2.914,P<0.05)。结论腺相关病毒介导B
Objective To investigate the effect of bone morphogenetic protein-9(BMP-9)mediated by adeno-associated virus on fracture healing in osteoporotic rats.Methods A total of 60 SD rats were randomly divided into sham group,model group and BMP-9 group.In the model group and BMP-9 group,osteoporosis model was established by removing bilateral ovaries,and osteoporotic fracture model was established by breaking the right femur 8 weeks later.In sham group,only the fat tissue around the ovary was removed.The rats in BMP-9 group were injected with 1×1010 vg of adeno-associated virus expressing BMP-9 at the fracture site,and those in sham group and model group were injected with 1×1010 vg of adeno-associated virus at the fracture site.After injection for 7 weeks.the recovery of fracture was scored by semi quantitative scoring method of X-ray and bone density was measured at fracture site.Mechanical parameters of fracture healing were measured by biomechanics.Changes of bone metabolism indexes were measured by enzyme linked immunosorbent assay(ELISA).The mRNA expression levels of vascular endothelial growth factor(VEGF)and nuclear factor-κB receptor activator ligand(RANKL)at fracture site were analyzed by fluorescence quantitative polymerase chain reaction(PCR).Results Compared with the model group(1.15±0.21),the level of BMP-9 mRNA in the fracture site of BMP-9 group significantly increased(3.94±0.34,t=3.089,P<0.05).Compared with the model group(2.23±0.25,0.077±0.002),the X-ray score and BMD in the BMP-9 group significantly increased(4.10±0.31,0.023±0.004,t=2.917,3.091,P<0.05).Compared with the model group,the biomechanical parameters of femur in BMP-9 group significantly improved(t=2.471,2.104,2.810,P<0.05).Compared with the model group(1.23±0.24,1.00±0.19),the level of VEGF and RANKL(2.58±0.31,3.29±0.22)in the BMP-9 group significantly increased(t=2.034,2.914,P<0.05).Conclusion The expression of BMP-9 mediated by adeno-associated virus can significantly improve bone metabolism,fracture healing and bone density.
作者
宋雨泽
李继峰
王晓
Song Yuze;Li Jifeng;Wang Xiao(Department of Orthopedics,Huaihe Hospital,Henan University,Kaifeng 475000,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2020年第6期1045-1048,共4页
Chinese Journal of Experimental Surgery
