盘龙七片调节Wnt/β-catenin通路对大鼠胫骨骨折愈合的影响

Effect of Panlongqi Tablet regulating Wnt/β-catenin pathway on the healing of tibial fracture in rats

目的探讨盘龙七片对大鼠胫骨骨折愈合的促进作用及可能作用机制。方法选用58只8周龄、SPF级SD雄性大鼠为实验大鼠,分为造模大鼠(n=48)与Sham组大鼠(n=10)。建立胫骨骨折模型大鼠,并将模型大鼠分为Model组、盘龙七片组(Treat组)、β-catenin抑制剂组(XAV-939)及盘龙七片+β-catenin抑制剂组(Treat+XAV-939组)4组,每组10只。X光片观察大鼠胫骨骨折愈合状况;骨密度测定仪检测胫骨骨矿物质密度值(BMD);HE染色观察大鼠骨折部位结构变化;酶联免疫吸附法检测血清中ALP、bFGF、TNF-α、IL-1β;免疫印迹法检测骨组织中p-糖原合成酶激酶3β(GSK-3β)、Wnt3、β-链蛋白(β-Catenin)及淋巴增强因子1(LEF1)蛋白表达。结果与Sham组相比,Model组骨组织影像学愈合评分、BMD、ALP、bFGF水平、Wnt3、β-Catenin、LEF1蛋白表达降低,TNF-α、IL-1β水平升高;与Model组相比,Treat组骨组织影像学愈合评分、BMD、ALP、bFGF水平、Wnt3、β-Catenin、LEF1蛋白表达升高,TNF-α、IL-1β水平降低;Treat+XAV-939组骨组织影像学愈合评分、BMD、ALP、bFGF水平、Wnt3、β-Catenin、LEF1蛋白表达高于XAV-939组,TNF-α、IL-1β水平低于XAV-939组,差异有统计学意义(均P<0.05)。结论盘龙七片可促进大鼠胫骨骨折愈合,其机制与抑制Wnt/β-catenin通路、缓解骨折大鼠机体炎症反应有关。

Objective To explore the promoting effect of Panlongqi Tablet on the healing of tibial fracture in rats and its possible mechanism.Methods The tibial fracture model rats were established and divided into Model group,Panlongqi Tablet group(Treat group),β-catenin inhibitor group(XAV-939),treat+XAV-939 group,with 10 rats in each group.The healing of tibial fracture was observed by X-ray.Bone mineral density(BMD)of tibia was measured by dual energy x-ray absorptiometry.HE staining was used to observe the structural changes of fracture site.ALP,bFGF,TNF-αand IL-1βin serum were detected by ELISA.Western blotting was used to detect the protein expressions of p-glycogen synthetase kinase 3β(GSK-3β),Wnt3,β-catenin and lymphoid enhancer 1(LEF1)in bone tissue.Results Compared with Sham group,the healing score of bone tissue imaging,BMD,the levels of ALP,bFGF,and the protein expressions of Wnt3,β-catenin and LEF1 in Model group were decreased,and the levels of TNF-αand IL-1βincreased.Compared with the Model group,the healing score of bone tissue imaging,BMD,the levels of ALP,bFGF,and the protein expressions of Wnt3,β-catenin and LEF1 in Model group were increased,and the levels of TNF-αand IL-1βdecreased.The healing score of bone tissue imaging,BMD,the levels of ALP,bFGF,and the protein expressions of Wnt3,β-catenin and LEF1 in Treat+XAV-939 group were higher than those in XAV-939 group,and the levels of TNF-αand IL-1βwere lower than those in XAV-939 group(P<0.05).Conclusion Panlongqi Tablet can promote the healing of tibial fracture in rats.Its mechanism is related to the inhibition of Wnt/β-catenin pathway and the alleviation of inflammatory response in rats.