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

作　　者：姑再阿依·买买提 唐卫东 蒲娟娟 张怀贵 GUZAIAYI Maimaiti;TANG Weidong(Xinjiang Uygur Autonomous Region People's Hospital,Xinjiang Urumqi 830000,China)

机构地区：[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

出　　处：《河北医学》2024年第9期1478-1483,共6页Hebei Medicine

基　　金：新疆维吾尔自治区自然科学基金资助项目,(编号:2023D01C417)。

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。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 healing 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 measuring 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.Surface markers CD9,CD63,and CD81 were prominently expressed.In the BMSCs-Exos group,miR-181 expression 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 

分 类 号：R683[医药卫生—骨科学]