以IGF-1、NOS、FGF23和硬骨素为靶基因的骨细胞力学响应微小RNA的筛选  

Screening of IGF-1,NOS,FGF23 and Sclerost-Targeted Mechanoresponsive MicroRNAs

作　　者：秦祎雄 曹振 杨焕 韩标 虞芳梅 胡婷婷[1] 郭勇[1] QIN Yixiong;CAO Zhen;YANG Huan;HAN Biao;YU Fangmei;HU Tingting;GUO Yong(Guangxi Higher Education key Laboratory of Biochemistry and Molecular Biology,College of Biotechnology,Guilin Medical University,Guilin 541100,China)

机构地区：[1]桂林医学院生物技术学院,广西高校生物化学与分子生物学重点实验室,桂林541100

出　　处：《医用生物力学》2021年第6期970-977,共8页Journal of Medical Biomechanics

基　　金：国家自然科学基金项目(31660261,32071309);广西自然科学基金项目(2018JIA140050)。

摘　　要：目的筛选骨细胞分泌因子相关力学响应微小RNA(microRNA,miRNA)。方法分别向体外培养骨细胞和成骨细胞施加周期性张应变(ε=2.5,f=0.5 Hz),采用miRNA芯片筛选出仅在骨细胞中差异表达的miRNA。通过生物信息学技术,从这些miRNA中进一步筛选出靶基因为胰岛素样生长因子1(insulin-like growth factor-1)、NO合成酶(nitric oxide synthesase,NOS)、成纤维细胞生长因子23(fibroblast growth factor 23,FGF23)和硬骨素(sclerostin,SOST)等分泌因子的miRNA,与小鼠跑台锻炼后芯片检测出的小鼠股骨组织差异表达miRNA进行比较,然后随机选4个miRNA进行定量PCR验证。结果仅在体外经力学刺激的骨细胞中差异表达的77个miRNA中,筛选出22个靶基因为4种分泌因子(IGF-1、NOS、FGF23、SOST)的miRNA,并进一步筛选出11个在跑台锻炼后的小鼠骨组织中以同样趋势差异表达的miRNA,其中随机选出的miR-361-3p、miR-3082-5p、miR-6348和miR-706,也在骨细胞和骨组织中以同样趋势同样差异表达。结论诸如miR-361-3p、miR-3082-5p、miR-6348和miR-706仅在骨细胞中差异表达的力学响应miRNA,很可能通过调节分泌因子影响成骨分化或骨代谢。Objective To screen the secretory factor-related,mechanoresponsive microRNAs(miRNA)of osteocytes.Methods Cyclic mechanical tensile strain(ε=2.5,f=0.5 Hz)was applied to osteocytes and osteoblasts cultured in vitro respectively,and the differentially expressed miRNAs only in the osteocytes were screened out by using miRNA chip.Through bioinformatics technology,in these differentially expressed miRNAs,the target genes of secretory factors including insulin-like growth factor-1(IGF-1),nitric oxide synthesase(NOS),fibroblast growth factor 23(FGF23)and sclerostin(SOST)were further screened out.Then the selected miRNAs were compared with the biochip detected,differentially expressed miRNAs in femur bone of the mice which were trained on treadmill,and four of these miRNAs were randomly selected for quantitative PCR verification.Results For the 77 differentially expressed miRNAs only in the mechanically strained osteocytes in vitro,22 miRNAs whose target genes were the 4 secreted factors(IGF-1,NOS,FGF23 and SOST),were screened out.Moreover,a total of 11 miRNAs in the 22 miRNAs were differentially expressed in femur bone of the treadmill trained mice with the same trend as those in osteocytes in vitro,and the randomly selected miR-361-3p,miR-3082-5p,miR-6348 and miR-706 were confirmed to be differentially expressed with the same trend in femur bone and osteocytes.Conclusions These mechanoresponsive miRNAs differentially expressed only in osteocytes,such as miR-361-3p,miR-3082-5p,miR-6348 and miR-706,probably influence osteoblastic differentiation or bone metabolism through regulating the secretory factors.

关 键 词：力学载荷 骨细胞 微小RNA 骨代谢 跑台运动 

分 类 号：R318.01[医药卫生—生物医学工程]