力学敏感性微RNA-337-3p调控骨髓间充质干细胞增殖和成骨分化介导脊柱废用性骨质疏松症发生的机制  

Mechanism of mechanosensitive microRNA-337-3p regulation of bone mesenchymal stem cell proliferation and osteogenic differentiation-mediated spinal disuse osteoporosis

作　　者：孙猛 董丰琴 廖腾 时国华 杨晓青 胡晓东 郭逸冰 于凤宾[1] Sun Meng;Dong Fengqin;Liao Teng;Shi Guohua;Yang Xiaoqing;Hu Xiaodong;Guo Yibing;Yu Fengbin(Department of Orthopaedics,72nd Group Army Hospital of Chinese PLA,Huzhou 313000,Zhejiang,China)

机构地区：[1]中国人民解放军陆军第72集团军医院骨科,湖州313000

出　　处：《脊柱外科杂志》2024年第6期379-385,391,共8页Journal of Spinal Surgery

基　　金：浙江省公益研究计划自然科学基金基础探索项目Y(LY20H060002);吴阶平医学基金会临床科研专项资助基金项目(320.6750.2021-04-54)。

摘　　要：目的探讨力学敏感性微RNA-337-3p(miR-337-3p)调控骨髓间充质干细胞(BMSC)增殖和成骨分化介导脊柱废用性骨质疏松症(DOP)发生的机制。方法分别给予人、小鼠、大鼠的BMSC机械牵张力学刺激并进行成骨诱导分化。采用聚合酶链式反应(PCR)检测各组BMSC中miR-337-3p的表达,通过抑制大鼠miR-337-3p的表达构建失重力模型。采用微计算机断层扫描技术(Micro-CT)检查大鼠骨量,评估miR-337-3p对DOP发生的影响。调控大鼠BMSC中miR-337-3p的表达,采用克隆增殖检测miR-337-3p对大鼠BMSC增殖的影响,采用碱性磷酸酶(ALP)染色、茜素红染色以及异位成骨分化检测miR-337-3p对大鼠BMSC成骨分化的影响。采用转录组测序和Western blotting寻找miR-337-3p的靶基因。结果机械牵张力学刺激抑制人、小鼠、大鼠来源的BMSC中miR-337-3p的表达,诱导BMSC成骨分化过程中各组miR-337-3p的表达显著降低。miR-337-3p敲除及功能抑制可缓解失重力引起的骨丢失,促进大鼠BMSC的增殖和成骨分化;过表达miR-337-3p可抑制大鼠BMSC的增殖和成骨分化。miR-337-3p抑制整合素β样蛋白1(ITGBL1)的表达。结论miR-337-3p通过抑制BMSC的增殖和分化介导脊柱DOP发生,抑制miR-337-3p的表达和功能可以缓解骨丢失。Objective To investigate the mechanism of mechanosensitive microRNA-337-3p(miR-337-3p)regulation of bone mesenchymal stem cell(BMSC)proliferation and osteogenic differentiation-mediated spinal disuse osteoporosis(DOP).Methods Mechanical traction stimulation was applied to BMSCs from humans,mice,and rats,and osteogenic differentiation was induced.Polymerase chain reaction(PCR)was used to detect the expression of miR-337-3p in BMSC of each group,and a gravity loss model was constructed by inhibiting the expression of miR-337-3p in rats.The bone mass of rats was examined by micro-computed tomography(Micro-CT)to evaluate the effect of miR-337-3p on the occurrence of DOP.The expression of miR-337-3p in rat BMSC was regulated,and the effect of miR-337-3p on the proliferation of rat BMSC detected by clone proliferation.The effect of miR-337-3p on osteogenic differentiation of rat BMSC was detected by alkaline phosphatase(ALP)staining,alizarin red staining and ectopic osteogenic differentiation.Target genes of miR-337-3p were identified by transcriptome sequencing and Western blotting.Results The expression of miR-337-3p from humans,mice,and rats BMSCs was inhibited by mechanical stretch stimulation,and induced a significant decrease in the expression of miR-337-3p in each group during the osteogenic differentiation process of BMSCs.Knockout and functional inhibition of miR-337-3p could alleviate bone loss caused by gravity loss,promote proliferation and osteogenic differentiation of rat BMSCs;overexpression of miR-337-3p could inhibit the proliferation and osteogenic differentiation of rat BMSCs.The expression of integrinβ-like protein 1(ITGBL1)was inhibited by miR-337-3p.Conclusion miR-337-3p mediates spinal DOP by inhibiting the proliferation and differentiation of BMSCs,and inhibition of the expression and function of miR-337-3p can alleviate bone loss.

关 键 词：微RNAS 间质干细胞 成骨细胞 细胞增殖 细胞分化 骨质疏松 

分 类 号：R681.4[医药卫生—骨科学]