BMSCs外泌体miR-181通过靶向调控ERK5促进股骨骨折小鼠的成骨分化
BMSCs Exosomes-Derived miR-181 Promotes Osteogenic Differentiation in Mice with Femoral Fracture by Targeting ERK5
姑再阿依·买买提 1唐卫东 1蒲娟娟 2张怀贵1
作者信息
- 1. 新疆维吾尔自治区人民医院整形外科,新疆 乌鲁木齐 830000
- 2. 新疆维吾尔自治区米东区人民医院儿科,新疆 乌鲁木齐 831400
- 折叠
摘要
目的:探讨BMSCs外泌体(BMSCs-Exos)对其体外成骨分化的调控作用,以及 miR-181通过靶向ERK5 实现的机制,并研究BMSCs-Exos和过表达 miR-181 对小鼠股骨骨折愈合的影响.方法:通过动态光散射(DLS)技术和Western blot法表征BMSCs-Exos,qRT-PCR和Western blot检测miR-181 和ERK5 表达,双荧光素酶报告基因实验验证 miR-181 对 ERK5 的调控.使用 Western blot 和ELISA检测蛋白表达和ALP 活性.在小鼠股骨骨折模型中注射 miR-181 mimic 或 BMSCs-Exos,通过骨痂体积(CV)、骨体积分数(BV/TV)和相关蛋白评估骨折愈合效果.结果:BMSCs-Exos 的粒径范围为50～150nm,Zeta 电势为-25.69±2.88mV,表面标志物 CD9、CD63、CD81 显著表达.BMSCs-Exos 组中miR-181 表达上调(P<0.05),ERK5 表达下调(P<0.05);mimic 上调了 miR-181 表达并降低 ERK5蛋白表达(P<0.05).BMSCs-Exos 转运 miR-181 通过靶向调控 ERK5 促进 BMSCs 体外成骨分化(P<0.05),且BMSCs-Exos和过表达miR-181 均促进小鼠股骨骨折愈合(P<0.05).结论:BMSCs-Exos 通过转运miR-181、靶向调控ERK5 促进BMSCs体外成骨分化,并有助于小鼠股骨骨折愈合.
Abstract
Objective:To investigate the regulatory effects of BMSCs-derived exosomes(BMSCs-Exos)on osteogenic differentiation in vitro,to elucidate the mechanism by which miR-181 targets ERK5 to mediate this regulation,and to study the impact of BMSCs-Exos and overexpressed miR-181 on femoral fracture heal-ing in mice.Methods:BMSCs-Exos were characterized using dynamic light scattering(DLS)and Western blot techniques.The expression levels of miR-181 and ERK5 were assessed via qRT-PCR and Western blot.Dual-luciferase reporter assays were performed to validate the targeting relationship between miR-181 and ERK5.Protein expression and ALP activity were analyzed using Western blot and ELISA.In a mouse femoral fracture model,miR-181 mimic or BMSCs-Exos were injected,and fracture healing was evaluated by meas-uring callus volume(CV),trabecular bone volume fraction(BV/TV),and relevant protein levels.Results:The particle size of BMSCs-Exos ranged from 50 to 150 nm,with a Zeta potential of-25.69±2.88 mV.Sur-face markers CD9,CD63,and CD81 were prominently expressed.In the BMSCs-Exos group,miR-181 ex-pression was upregulated(P<0.05),while ERK5 expression was downregulated(P<0.05).The miR-181 mimic increased miR-181 expression and decreased ERK5 protein levels(P<0.05).BMSCs-Exos delivering miR-181 promoted osteogenic differentiation of BMSCs in vitro by targeting ERK5(P<0.05).Both BMSCs-Exos and overexpressed miR-181 enhanced femoral fracture healing in mice(P<0.05).Conclusion:BMSCs-Exos facilitate osteogenic differentiation of BMSCs in vitro by transporting miR-181 and targeting ERK5,and they also aid in femoral fracture healing in mice.
关键词
骨髓间充质干细胞外泌体/微小RNA-181/细胞外信号调节激酶5/股骨骨折小鼠/成骨分化Key words
Bone marrow mesenchymal stem cell-derived exosomes/microRNA-181/Extracel-lular signal-regulated kinase 5/Mouse femur fracture/Osteogenic differentiation引用本文复制引用
基金项目
新疆维吾尔自治区自然科学基金资助项目(2023D01C417)
出版年
2024
国家科技期刊平台
NSTL
万方数据