微小RNA-126对大鼠骨痂组织血管生成的影响  

Effects of microRNA-126 on angiogenesis in rat callus tissue

作　　者：廉檬檬 聂云飞[1] 李放 田翔宇[1] Lian Mengmeng;Nie Yunfei;Li Fang;Tian Xiangyu(Department of Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院外科医学部,450052

出　　处：《中华实验外科杂志》2022年第4期697-700,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探究微小RNA(miR)-126对骨折愈合中血管生成的影响。方法构建股骨骨折大鼠模型,局部注射miR-126激动剂(agomiR-126)、miR-126拮抗剂(antagomiR-126)设立实验组,同时设立对照组(agomiR-NC),于术后4、8、12周通过显微计算机断层扫描(micro-CT)检测各组股骨骨折愈合,通过免疫组织化学标记骨痂内血管内皮标志物CD31,并计算微血管密度(MVD)值,采用蛋白质印迹法(Western blot)及反转录-聚合酶链反应(RT-PCR)分别检测骨痂中血管生成相关因子出芽相关蛋白(SPRED1)、血管内皮生长因子A(VEGFA)的蛋白及mRNA水平,两组间比较采用t检验。结果micro-CT结果显示,在术后12周agomiR-126组右股骨骨折线基本消失,其骨折局部的骨密度[(732.1±16.3)mg/cm]及BV/TV比值[(72.4±2.1)%]明显高于对照组[(684.2±13.2)mg/cm、(70.3±2.3)%],差异有统计学意义(t=11.517,t=9.261,P<0.01);而antagomiR-126组骨折间隙仍较为明显,其骨密度[(635.2±12.6)mg/cm]及BV/TV比值[(65.1±3.4)%]明显下降(t=8.174,t=6.838,P<0.05)。免疫组织化学结果显示,agomiR-126组骨痂中MVD值(7.6±2.1)明显高于对照组(5.6±1.2),差异有统计学意义(t=4.180,P<0.01),而antagomiR-126组中MVD值(3.6±1.1)则明显下降(t=3.725,P<0.01)。Western blot及RT-PCR结果显示,agomiR-126组SPRED-1蛋白及mRNA表达水平(0.47±0.04、0.68±0.36)均明显下降(t=5.412、4.712,P<0.01),其VEGFA蛋白及mRNA表达水平(0.68±0.02、2.53±0.79)则明显上升(t=5.284、11.625,P<0.01);相反,antagomiR-126组SPRED-1蛋白及mRNA表达水平(0.87±0.06、1.65±0.37)均明显上升(t=7.223、6.795,P<0.01),其VEGFA蛋白及mRNA表达水平(0.43±0.04、0.75±0.29)则明显下降(t=4.226、2.169,P<0.05)。结论miR-126可以通过SPRED-1/VEGFA通路促进大鼠股骨骨痂组织中的血管生成,并加快骨折的愈合。Objective To explore the effect of microRNA(miR)-126 on angiogenesis in fracture healing.Methods A rat model of femoral fracture was constructed,and an experimental group was established by local injection of agomiR-126 and antagomiR-126,respectively,and agomiR-NC served as a control group.The femoral fracture healing of each group was detected by micro-computed tomography(micro-CT)at 4,8 and 12 weeks after operation.The vascular endothelial marker CD31 in the callus was marked by immunohistochemistry,and the microvessel density(MVD)value was calculated.Western blotting and reverse transcriptase-polymerase chain reaction(RT-PCR)were used to detect the protein and mRNA levels of angiogenesis-related factors SPRED1 and vascular endothelial growth factor A(VEGFA),respectively.T test was used for comparison between two groups.Results The micro-CT results showed that at 12 weeks after operation,the fracture line of the right femur of the agomiR-126 group basically disappeared,and the bone mineral density[(732.1±16.3)mg/cm]and BV/TV ratio[(72.4±2.1)%]of the fracture site were significantly higher than those of the control group[(684.2±13.2)mg/cm,(70.3±2.3)%],with statistical differences(t=11.517,t=9.261,P<0.01).However,the fracture gap was still obvious in the antagomiR-126 group,and its bone mineral density[(635.2±12.6)mg/cm]and BV/TV ratio[(65.1±3.4)%]were significantly lower(t=8.174,t=6.838,P<0.05).The immunohistochemical results showed that the MVD value(7.6±2.1)in bone callus was significantly higher than that in the control group(5.6±1.2),with statistical significance(t=4.180,P<0.01).The MVD value in the antagomiR-126 group(3.6±1.1)was significantly lower(t=3.725,P<0.01).Western blotting and RT-PCR results showed that the expression levels of SPRED-1 protein and mRNA in agomiR-126 group(0.47±0.04,0.68±0.36)were significantly decreased(t=5.412,t=4.712,P<0.01),while the expression levels of VEGFA protein and mRNA(0.68±0.02,2.53±0.79)were significantly increased(t=5.284,11.625,P<0.01).On the contrary,th

关 键 词：微小RNA 血管生成 骨折愈合 

分 类 号：R683[医药卫生—骨科学]