微小RNA-151-5p通过转化生长因子-β通路参与骨折愈合的早期调控过程  

MicroRNA-151-5p involves in the early healing of fractures via regulating the transforming growth factor-β signaling pathway

作　　者：赵亮[1] 李金峰[1] 杨浩[1] 乔志[1] 宋瑞鹏[1] 王卫东[1] 刘子汭 王利民[1] Zhao Liang;Li Jinfeng;Yang Hao;Qiao Zhi;Song Ruipeng;Wang Weidong;Liu Zirui;Wang Limin(Department of Orthopedics,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 45052,China)

机构地区：[1]郑州大学第一附属医院骨科,郑州450052

出　　处：《中华实验外科杂志》2023年第1期115-118,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨微小RNA(miRNA,miR)-151-5p在骨折愈合方面的作用。方法建立标准的大鼠股骨干骨折愈合模型(郑州大学动物中心),基因芯片分析血浆miRNA的表达。应用生物信息学方法模拟miRNA与靶基因配对,并寻找相关的靶通路。细胞实验验证miR-151-5p对成骨细胞(大鼠颅骨提取)的影响。最后,再次建立骨折愈合模型,给予miR-151-5p模仿物和抑制物,利用酶联免疫吸附试验(ELISA)检测转化生长因子-β(TGF-β)通路主要蛋白的表达。正态分布数据应用student-t检验,miRNA数据应用Benjamini&Hochberg方法进行比对。结果在大鼠骨折模型血浆结果显示miR-151-5p在骨折愈合7 d时显著高于对照组(对照比7 d为58.16±8.17比201.40±7.23,F=25.09,P<0.01),GSE93083数据集中骨折患者miRNA-151-5p表达量较健康组明显升高(骨折患者比健康人为12.25±0.17比12.00±0.19,t=2.446,P<0.01)。通过生物信息学分析,miR-151-5p的靶基因参与丝裂原活化蛋白激酶(MAPK)信号通路,胰岛素信号通路,上皮细胞间质转型(EMT)信号通路,Ras信号通路以及TGF-β信号通路的调控。文献分析富集结果提示,miR-151-5p的靶基因主要参与TGF-β信号通路的调控。细胞实验提示miR-151-5p可以抑制成骨细胞增值活性,并抑制成骨细胞TGF-β通路表达。骨折愈合模型中,miR-151-5p模仿物PLCG1(miR-151-5p模仿物比抑制物组为3.99±2.06比10.36±2.87,q=4.596,P<0.05)和MAPK8(miR-151-5p模仿物比抑制物组为4.93±1.543比17.49±5.328,q=4.533,P<0.01)较抑制物组显著增高。结论miR-151-5p是通过TGF-β参与骨折愈合的早期调控过程。Objective To explore the effect and mechanism of microRNA(miRNA,miR)-151-5p on early fracture healing.Methods A rat femoral shaft fracture healing model was established and the expression of miRNA in plasma was analyzed.The bioinformatics analysis was used to simulate the pairing of miRNA and target gene.Cell experiments verified the effect of miR-151-5p on osteoblasts(extracted from rat skull).Finally,the fracture healing model was established and miR-151-5p mimics and inhibitors were given to detect the expression of major proteins in transforming growth factor-β(TGF-β)pathway.Student-t test and Benjamini&Hochberg were used for data analysis.Results Animal experiments showed that miR-151-5p in rat plasma was significantly higher at the 7th day of fracture healing(control vs.7 d:58.16±8.17 vs.201.40±7.23,F=25.09,P<0.01),and the expression of miR-151-5p in fracture patients from GEO database was significantly higher than that in healthy group(fracture patients vs.healthy people:12.25±0.17 vs.12.00±0.19,t=2.446,P<0.01).The target genes of miR-151-5p were involved in mitogen activated protein kinase,insulin signaling pathway,epithelial mesenchymal transition signaling pathway,and Ras signaling pathway.The enrichment results of literature analysis suggest that the target gene of miR-151-5p was mainly involved in the regulation of TGF-β signaling pathway.Cell experiments suggested that miR-151-5p inhibited the proliferation of osteoblasts and down-regulated the expression of TGF-β pathway in osteoblasts.In the fracture healing model,the miR-151-5p mimic PLCG1(miR-151-5p mimic vs.inhibitor group:3.99±2.06 vs.10.36±2.87,q=4.596,P<0.05)and MAPK8(miR-151-5p mimics vs.inhibitors group:4.93±1.543 vs.17.49±5.328,q=4.533,P<0.01)were significantly higher than inhibitors group.Conclusion MiR-151-5p participates in the early regulatory process of fracture healing through the TGF-β signaling pathway.

关 键 词：骨折愈合 微小RNA 转化生长因子-β信号通路 

分 类 号：R683[医药卫生—骨科学]