激活素A在小鼠股骨骨折愈合中的作用实验研究  

Role of activin A in femoral fracture healing in mice

作　　者：刘方[1] 李文庆[2] 申晟[2] 张耘[1] LIU Fang;LI Wenqing;SHEN Sheng;ZHANG Yun(Department of Hand Surgery,Luoyang Orthopedic-Traumatological Hospital of Henan Province,Luoyang 471002,China)

机构地区：[1]河南省洛阳正骨医院手外科,河南洛阳471002 [2]河南省洛阳正骨医院脊柱科,河南洛阳471002

出　　处：《陕西医学杂志》2024年第12期1617-1623,共7页Shaanxi Medical Journal

基　　金：河南省中医药科学研究专项课题(2024ZY1019)。

摘　　要：目的:探讨激活素A在小鼠股骨骨折愈合中的作用。方法:选取8周龄骨发育成熟的雄性C57BL/6小鼠60只进行骨折造模后,随机分为对照组和激活素A组(给予激活素A治疗),每组30只。通过X线检查评估骨折愈合情况,采用微型计算机断层扫描(μCT)分析骨痂形成情况。采用激活素A处理人脐静脉内皮细胞(HUVECs)和骨髓间充质干细胞(BMSCs),通过细胞迁移实验、碱性磷酸酶(ALP)染色、茜素红染色和实时定量聚合酶链反应(RT-qPCR)分析激活素A对成骨分化和血管生成的促进作用。结果:在15、18、21 d,激活素A组骨折愈合评分高于对照组,且在21 d时激活素A组评分最高(均P<0.05)。术后1、2、3周,与对照组比较,激活素A组骨痂骨体积和连接密度增加,骨小梁分离度减小(均P<0.05)。激活素A组血管表面定量、血管数量和血管体积分数在术后第2周高于对照组(均P<0.05)。术后1、2周,激活素A组血管内皮生长因子A(VEGFA)阳性率高于对照组,术后3周则低于对照组(均P<0.05)。两组术后1、2、3周成骨分化基因骨形态发生蛋白2(BMP2)阳性率比较差异无统计学意义(均P>0.05)。ALP染色结果显示,成骨标志物ALP表达在激活素A处理1周后增加。茜素红染色结果显示,激活素A组矿化区域较对照组多。RT-qPCR结果显示,处理1、2、3周后,激活素A组BMSCs细胞ALP、Sp7转录因子(OSX)、BMP2 mRNA表达高于对照组(均P<0.05)。与对照组比较,激活素A组HUVECs细胞12、24 h细胞迁移率增加,VEGFA、CD31、成纤维细胞生长因子2(FGF2)mRNA表达增加(均P<0.05)。结论:激活素A可以上调BMSCs和HUVECs细胞中BMP2和VEGFA基因的表达,促进细胞成骨分化和血管生成,从而加快骨折骨愈合速度。Objective:To investigate the role of activin A in femoral fracture healing in mice.Methods:A total of 608-week-old male C57BL/6 mice with mature bones were selected for fracture modeling and randomly divided into control group and activin A group(treated with activin A),with 30 mice in each group.Fracture healing was analyzed by X-ray,and bone scab formation was analysed byμCT.HUVECs and BMSCs were treated with activin A,and the promoting effects of activin A on osteogenic differentiation and angiogenesis were analyzed by cell migration assay,ALP staining,alizarin red staining and RT-qPCR.Results:At 15,18 and 21 days,the fracture healing scores of activin A group were higher than those of the control group,and the score of activin A group was the highest at 21 days(all P<0.05).Compared with the control group,the volume and connection density of bone scab were increased,and the separation of trabecular bone was decreased in the activin A group at 1,2 and 3 weeks after operation(all P<0.05).Vessel surface quantification,vessel number and vessel volume fraction in activin A group were higher than those in control group at 2 weeks(all P<0.05).The positive rate of VEGFA in activin A group was higher than that in control group at 1 and 2 weeks after operation,and lower than that in control group at 3 weeks after operation(all P<0.05).There was no significant difference in the positive rate of BMP2 between the two groups at 1,2 and 3 weeks after operation(all P>0.05).ALP staining results showed that the expression of osteogenic marker ALP increased after 1 week of activin A treatment.Alizarin red staining showed that the activin A group had more mineralized areas than the control group.RT-qPCR results showed that the mRNA expressions of ALP,OSX and BMP2 in activin A group were higher than those in control group in BMSCs after 1,2 and 3 weeks of treatment(all P<0.05).Compared with the control group,the migration rate of HUVECs cells in activin A group was increased at 12 and 24 hours,and the mRNA expressions of VEGFA,CD

关 键 词：激活素A 骨折愈合 血管生成 血管内皮生长因子A 骨形态发生蛋白2 小鼠 

分 类 号：R-33[医药卫生]