贵州医科大学学报

2021, v.46;No.254(11) 1309-1315

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印度刺猬蛋白基因敲除对腓骨骨折小鼠愈合的影响
The effect of indian hedgehog gene knockout on the healing of fibula fracture in mice

刘超;张焱祥;王显勋;李勇光;
LIU Chao;ZHANG Yanxiang;WANG Xianxun;LI Yongguang;Department of Orthopedics,Hubei No.3 People's Hospital of Jianghan University;

摘要(Abstract):

目的探讨印度刺猬蛋白(Ihh)基因敲除对腓骨骨折小鼠骨愈合的影响。方法将40只刚出生、基因型为Ihhf1/f1和Col2a1-Cre ERT2的小鼠随机均分为观察组和对照组,观察组小组诱导敲除Ihh基因,对照组不敲除;2组小鼠均制作腓骨骨折模型,于造模后第1、2、3周末时,X线观察小数腓骨骨折线,骨折部位行Micro-CT扫描,同时取血检测血清血管内皮生长因子(VEGF)、转化生长因子(TGF-β1)、碱性磷酸酶(ALP)、骨钙素(BGP)、钙与磷离子浓度、钙磷乘积,取同时点腓骨样本行ABH染色。结果造模后第2周开始,2组小鼠均有模糊骨折线、且骨性骨痂连接影增多,观察组小鼠的骨折线非常模糊的比例较对照组降低(P<0.05);造模后第2周和第3周时、2组小鼠的结构模型指数(SMI)、组织体积(TV)、骨体积(BV)、骨体积分数(BV/TV)、骨小梁数量(Tb.N)及血清VEGF、TGF-β1、ALP、BGP水平均较第1周降低(P<0.05),而骨小梁模式因子(Tb.Pf)、骨小梁厚度(Tb.Th)及血清钙离子浓度均升高(P<0.05);与造模后第1周和第2周时比较,造模3周时观察组仅出现少量骨性骨痂,骨小梁小于对照组,且形状不规则,而对照组的骨小梁形状较观察组规则、粗大。结论 Ihh基因敲除会抑制腓骨骨折小鼠血管增生,降低钙磷水平,进而影响骨性骨痂的生成,延缓骨折的愈合。
Objective To investigate the effect of Indian hedgehog(Ihh) gene knockout on bone healing in mice with fibula fractures.Methods Forty newborn mice with genotypes of Ihh(fl/fl) and Col2 a1-Cre ERT2 were randomly divided into observation group and control group.The observation group was induced to knock out the Ihh gene while the control group was not.In both groups of mice,the fibula fracture model was made.On the 1(fl/fl) and Col2 a1-Cre ERT2 were randomly divided into observation group and control group.The observation group was induced to knock out the Ihh gene while the control group was not.In both groups of mice,the fibula fracture model was made.On the 1(st),2(st),2(nd),and 3(nd),and 3(rd) weekend after the model establishment,X-ray was used to observe the fibula fracture line and Micro-CT parameters at the fracture site.Get blood sample to check serum vascular endothelial growth factor(VEGF),transforming growth factor(TGF-β1),alkaline phosphatase(ALP),osteocalcin(BGP),calcium and phosphorus ion concentration,and calcium-phosphorus product.The fibula samples were taken for ABH staining.Results Two weeks after the modeling,the mice in the two groups had fuzzy fracture lines and the bony callus was more visible.The fuzzy degree of fracture line in the observation group was lower than that in the control group.On 2(rd) weekend after the model establishment,X-ray was used to observe the fibula fracture line and Micro-CT parameters at the fracture site.Get blood sample to check serum vascular endothelial growth factor(VEGF),transforming growth factor(TGF-β1),alkaline phosphatase(ALP),osteocalcin(BGP),calcium and phosphorus ion concentration,and calcium-phosphorus product.The fibula samples were taken for ABH staining.Results Two weeks after the modeling,the mice in the two groups had fuzzy fracture lines and the bony callus was more visible.The fuzzy degree of fracture line in the observation group was lower than that in the control group.On 2(nd) and 3(nd) and 3(rd) week after modeling,the structural model index(SMI),tissue volume(TV),bone volume(BV),bone volume fraction(BV/TV),Trabecular Number(Tb.N),as well as serum VEGF,TGF-β1,ALP,and BGP levels in both groups of mice decreased,compared with those in the 1(rd) week after modeling,the structural model index(SMI),tissue volume(TV),bone volume(BV),bone volume fraction(BV/TV),Trabecular Number(Tb.N),as well as serum VEGF,TGF-β1,ALP,and BGP levels in both groups of mice decreased,compared with those in the 1(st) week(P <0.05).In contrast,trabecular bone model factor(Tb.Pf),trabecular bone thickness(Tb.Th) and serum calcium ion concentration increased(P <0.05).Compared with 1(st) week(P <0.05).In contrast,trabecular bone model factor(Tb.Pf),trabecular bone thickness(Tb.Th) and serum calcium ion concentration increased(P <0.05).Compared with 1(st) and 2(st) and 2(nd) week after modeling,the 3(nd) week after modeling,the 3(rd) week saw only a small amount of bony callus in the observation group,and the trabecular bone was irregularly shaped and was significantly smaller than that of the control group.The shape of trabecular bone in the control group,by contrast,was regular and thicker than that of the observation group.Conclusion Ihh gene knockout inhibits vascular proliferation in mice with fibula fractures,reduces calcium and phosphorus levels,thereby affecting the formation of bony callus and delaying the healing of fractures.

关键词(KeyWords): 腓骨骨折;小鼠;Ihh基因敲除;骨折线;骨性骨痂;愈合
fibula fracture;mice;Ihh gene knockout;fracture line;bony callus;heal

Abstract:

Keywords:

基金项目(Foundation): 武汉市卫生和计划生育委员会科研项目(WX18C33);; 湖北省卫生健康委员会科研项目(WJ2019M248)

作者(Authors): 刘超;张焱祥;王显勋;李勇光;
LIU Chao;ZHANG Yanxiang;WANG Xianxun;LI Yongguang;Department of Orthopedics,Hubei No.3 People's Hospital of Jianghan University;

DOI: 10.19367/j.cnki.2096-8388.2021.11.011

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