[1]刘 方,李文庆,申 晟,等.激活素A在小鼠股骨骨折愈合中的作用实验研究[J].陕西医学杂志,2024,(12):1617-1623.[doi:DOI:10.3969/j.issn.1000-7377.2024.12.006]
 LIU Fang,LI Wenqing,SHEN Sheng,et al.Role of activin A in femoral fracture healing in mice[J].,2024,(12):1617-1623.[doi:DOI:10.3969/j.issn.1000-7377.2024.12.006]
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激活素A在小鼠股骨骨折愈合中的作用实验研究
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
期数:
2024年12期
页码:
1617-1623
栏目:
基础研究
出版日期:
2024-12-05

文章信息/Info

Title:
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
(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)
Author(s):
LIU FangLI WenqingSHEN ShengZHANG Yun
(Department of Hand Surgery,Luoyang Orthopedic-Traumatological Hospital of Henan Province,Luoyang 471002,China)
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
Keywords:
Activin A Fracture healing Angiogenesis Vascular endothelial growth factor A Bone morphogenetic protein 2 Mouse
分类号:
R -33
DOI:
DOI:10.3969/j.issn.1000-7377.2024.12.006
文献标志码:
A
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。
Abstract:
Objective:To investigate the role of activin A in femoral fracture healing in mice.Methods:A total of 60 8-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 by μCT.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,CD31 and FGF2 wwere increased(all P<0.05).Conclusion:Activin A can up-regulate the expression of BMP2 and VEGFA genes in BMSCs and HUVECs,promote cell osteogenic differentiation and angiogenesis,and thus accelerate fracture healing.

参考文献/References:

[1] WILDEMANN B,IGNATIUS A,LEUNG F,et al.Non-union bone fractures[J].Nat Rev Dis Primers,2021,7(1):57.
[2] EKEGREN C L,EDWARDS E R,DE STEIGER R,et al.Incidence,costs and predictors of non-union,delayed union and mal-union following long bone fracture[J].Int J Environ Res Public Health,2018,15(12):2845.
[3] SUN X,LI P,QU X,et al.Isovitexin alleviates acute gouty arthritis in rats by inhibiting inflammation via the TLR4/MyD88/NF-κB pathway[J].Pharm Biol,2021,59(1):1326-1333.
[4] DING Z C,LIN Y K,GAN Y K,et al.Molecular pathogenesis of fracture nonunion[J].J Orthop Translat,2018,14:45-56.
[5] EINHORN T A,GERSTENFELD L C.Fracture healing:Mechanisms and interventions[J].Nat Rev Rheumatol,2015,11(1):45-54.
[6] STEGEN S,VAN GASTEL N,CARMELIET G.Bringing new life to damaged bone:The importance of angiogenesis in bone repair and regeneration[J].Bone,2015,70:19-27.
[7] 卢承印,罗志强,简功辉,等.H型血管促进成血管-成骨耦联在骨折愈合中作用机制的研究进展[J].华中科技大学学报(医学版),2024,53(1):133-139.
[8] 冯亮,樊力.骨髓间充质干细胞通过增加血管生成和血管内皮生长因子表达影响大鼠随意型皮瓣存活实验研究[J].陕西医学杂志,2022,51(3):298-302.
[9] 李润媛,黄于庭.胃癌患者化疗前后血管内皮生长因子动态变化与临床获益关系研究[J].陕西医学杂志,2023,52(8):1042-1046.
[10] TAKAGI K,URIST M R.The reaction of the dura to bone morphogenetic protein(BMP)in repair of skull defects[J].Ann Surg,1982,196(1):100-109.
[11] CHEN X,TAN B,BAO Z,et al.Enhanced bone regeneration via spatiotemporal and controlled delivery of a genetically engineered BMP-2 in a composite hydrogel[J].Biomaterials,2021,277:121117.
[12] 崔书伟,高小康,刘晓宁.沉默微小RNA-378靶向调节骨形态发生蛋白2/母亲DPP同源物4信号通路对大鼠胫骨骨折愈合的影响实验研究[J].陕西医学杂志,2024,53(6):748-753.
[13] SAKURAI T,ABE Y,KASUYA Y,et al.Activin A stimulates mitogenesis in Swiss 3T3 fibroblasts without activation of mitogen-activated protein kinases[J].J Biol Chem,1994,269(19):14118-14122.
[14] CENTRELLA M,MCCARTHY T L,CANALIS E.Activin A binding and biochemical effects in osteoblast-enriched cultures from fetal-rat parietal bone[J].Mol Cell Biol,1991,11(1):250-258.
[15] FERGUSON V L,AYERS R A,BATEMAN T A,et al.Bone development and age-related bone loss in male C57BL/6J mice[J].Bone,2003,33(3):387-398.
[16] YUASA M,SAITO M,MOLINA C,et al.Unexpected timely fracture union in matrix metalloproteinase 9 deficient mice[J].PLoS One,2018,13(5):e0198088.
[17] KHAN S N,LANE J M.The use of recombinant human bone morphogenetic protein-2(rhBMP-2)in orthopaedic applications[J].Expert Opin Biol Ther,2004,4(5):741-748.
[18] LO K W,ULERY B D,ASHE K M,et al.Studies of bone morphogenetic protein-based surgical repair[J].Adv Drug Deliv Rev,2012,64(12):1277-1291.
[19] YAO L,LU J,ZHONG L,et al.Activin A marks a novel progenitor cell population during fracture healing and reveals a therapeutic strategy[J].Elife,2023,12:e89822.
[20] ZHU D,FANG H,YU H,et al.Alcohol-induced inhibition of bone formation and neovascularization contributes to the failure of fracture healing via the miR-19a-3p/FOXF2 axis[J].Bone Joint Res,2022,11(6):386-397.
[21] KLEINHEINZ J,STRATMANN U,JOOS U,et al.VEGF-activated angiogenesis during bone regeneration[J].J Oral Maxillofac Surg,2005,63(9):1310-1316.
[22] STREET J,BAO M,DEGUZMAN L,et al.Vascular endothelial growth factor stimulates bone repair by promoting angiogenesis and bone turnover[J].Proc Natl Acad Sci USA,2002,99(15):9656-9661.
[23] GERBER H P,VU T H,RYAN A M,et al.VEGF couples hypertrophic cartilage remodeling,ossification and angiogenesis during endochondral bone formation[J].Nat Med,1999,5(6):623-628.
[24] WEI X,WANG J,DENG Y Y,et al.Tubiechong patching promotes tibia fracture healing in rats by regulating angiogenesis through the VEGF/ERK1/2 signaling pathway[J].J Ethnopharmacol,2023,301(1):115851.
[25] DE OLIVEIRA C E,DOURADO M R,SAWAZAKI-CALONE I,et al.Activin A triggers angiogenesis via regulation of VEGFA and its overexpression is associated with poor prognosis of oral squamous cell carcinoma[J].Int J Oncol,2020,57(1):364-376.
[26] LI Y,ZHU H,KLAUSEN C,et al.Vascular endothelial growth factor-A(VEGF-A)mediates activin A-induced human trophoblast endothelial-like tube formation[J].Endocrinology,2015,156(11):4257-4268.

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备注/Memo

备注/Memo:
基金项目:河南省中医药科学研究专项课题(2024ZY1019)
更新日期/Last Update: 2024-12-05