TNF-α对人骨髓间充质干细胞sncRNAs表达的调控研究

A study on the regulation of tumor necrosis factorαon the expression of small non-coding ribonucleic acids in human bone marrow mesenchymal stem cells

目的探讨肿瘤坏死因子α(TNF-α)对人骨髓间充质干细胞(hBMSCs)中非编码小RNA(sncRNAs)表达的调控作用。方法选择2021年8月至2021年10月于上海交通大学医学院附属新华医院骨科接受手术治疗的先天性髋关节发育不良患者5例。通过全骨髓血贴壁法分离hBMSCs。TNF-α组采用含有10 ng/ml TNF-α的完全培养基进行干预,对照组加入等体积的完全培养基。干预24 h后提取hBMSCs总RNA,应用small RNA测序法检测sncRNAs的表达情况,并应用实时荧光定量聚合酶链反应(PCR)进行验证。结果经TNF-α干预,hBMSCs中5个miRNA(hsa-novel-170-mature、hsa-miR-155-5p、hsa-miR-1260a、hsa-novel-202-mature、hsa-novel-116-mature)、4个piRNA(DQ592932、DQ570992、DQ593325、DQ582827)的表达上调(P<0.05),4个miRNA(hsa-miR-141-3p、hsa-miR-3065-5p、hsa-novel-155-mature、hsa-miR-27a-5p)、13个snoRNA(SNORD14D、SNORD116-3、SNORA21、SNORD60、SNORD81、SNORD32A、SNORD117、SNORD2、SNORD1C、SNORD36B、SNORD4B、SNORA58、SNORA64)和1个snRNA(NR_003137.2)的表达下调(P<0.05)。结论TNF-α可以调控hBMSCs中多种sncRNAs的表达,进而影响hBMSCs成骨分化过程。

Objective To investigate the regulatory effect of tumor necrosis factorα(TNF-α)on the expression of small non-coding ribonucleic acids(sncRNAs)in human bone marrow mesenchymal stem cells(hBMSCs).MethodsFive patients with congenital hip dysplasia who received surgical treatment in Department of Orthopedics,Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from August 2021 to October 2021 were selected.hBMSCs were isolated by whole bone marrow blood adherence method.The TNF-αgroup was intervened with complete medium containing 10 ng/ml TNF-α,and the control group was added with an equal volume of complete medium.The total RNA of hBMSCs was extracted after 24 hours of intervention,and the expression of sncRNAs was detected by small RNA sequencing,and verified by real-time quantitative polymerase chain reaction(PCR).Results After intervention with TNF-α,the expressions of 5 micro-ribonucleic acids(miRNAs)(hsa-novel-170-mature,hsa-miR-155-5p,hsa-miR-1260a,hsa-novel-202-mature,hsa-novel-116-mature)and 4 PIWI-interacting RNAs(piRNAs)(DQ592932,DQ570992,DQ593325,DQ582827)in hBMSCs were up-regulated(P<0.05),and 4 miRNAs(hsa-miR-141-3p,hsa-miR-3065-5p,hsa-novel-155-mature,hsa-miR-27a-5p),13 small nucleolar RNAs(snoRNAs)(SNORD14D,SNORD116-3,SNORA21,SNORD60,SNORD81,SNORD32A,SNORD117,SNORD2,SNORD1C,SNORD36B,SNORD4B,SNORA58,SNORA64)and 1 small nuclear RNA(snRNA)(NR_003137.2)were down-regulated(P<0.05).Conclusion TNF-αcan regulate the expressions of various sncRNAs in hBMSCs,thereby affecting the osteogenic differentiation of hBMSCs.