摘要
目的探讨微小RNA(miR)-125b-5p对骨质疏松症(OP)大鼠骨丢失的影响及其分子机制。方法从公共基因表达数据库(GEO)获取OP相关数据集筛选差异基因,分析OP患者与健康人miR-125b-5p表达量的差异;通过切除大鼠双侧卵巢构建OP大鼠模型;6只健康清洁级Sprague-Dawley(SD)大鼠均分为OP组(切除大鼠双侧卵巢)和假手术组(只行背部切口,不切除卵巢;Sham组),术后3个月采用显微计算机断层扫描(micro-CT)评价OP模型构建是否成功;另取9只雌性SD大鼠切除双侧卵巢,随机均分为空载体组(agomiR-NC组,agomiR-NC干粉33μg+PBS溶液125μL)、磷酸缓冲溶液(PBS)组(PBS溶液125μL)及miR-125b-5p模拟剂(agomiR-125b-5b组,agomiR-125b-5p干粉33μg+PBS溶液125μL),尾静脉注射,1次/周、持续2个月,干预结束后采用micro-CT扫描各组大鼠胫骨,收集骨体积分数(BV/TV)、骨密度(BMD)、骨小梁数目(Tb.N)、骨小梁分离度(Tb.Sp)及结构模式指数(SMI)等骨参数;采用生物信息学技术对miR-125b-5p进行靶基因预测、基因本体(GO)功能和京都基因和基因基因组百科全书(KEGG)信号通路富集分析。结果与健康人群比较,OP患者血浆中有差异miRNA 956个,其中上调502个、下调454个,尤以miR-125b-5p表达升高明显;OP组大鼠胫骨BV/TV及BMD低于Sham组(P<0.05或P<0.01),提示骨质疏松模型造模成功;与agomiR-NC组比较,agomiR-125b-5p组大鼠胫骨BV/TV、Tb.N降低,Tb.Sp、SMI升高(P<0.05),但PBS组与agomiR-NC组大鼠胫骨上述骨参数比较、差异无统计学意义(P>0.05);4种软件及5种软件预测显示miR-125b-5p的靶基因分别为102个、8个,生物过程(BP)主要为pre-microrna的处理、负调控冷诱导产热及调节细胞生长的程度等,分子功能(MF)主要为金属胺肽酶活性、泛素结合酶活性等,KEGG信号通路主要为肾素-血管紧张素系统和泛素介导的蛋白质水解等。结论miR-125b-5p过表达可促进OP大鼠的骨丢失,其机制可能与调节BCL2拮抗剂/杀伤�
Objective To investigate the effects of microRNA(miR)-125b-5p on bone loss of rats with osteoporosis(OP)and its molecular mechanism.Methods Op-related data sets were obtained from the public gene expression database(GEO)to screen differential genes,and the differences of miR-125b-5p expression between OP patients and healthy people were analyzed.The OP rat model was constructed by resecting bilateral ovaries.Six healthy and clean Sprague-Dawley(SD)rats were divided into two groups:OP group(resection of bilateral ovaries)and the sham group(resection of back without ovaries).In Sham group,micro-computed tomography(micro-CT)was used 3 months after operation to evaluate the success of OP model construction.Other 9 female SD rats underwent bilateral ovaries resection and were randomly divided into the empty vector group(agomiR-NC group,agomiR-NC dry powder 33μg+PBS solution 125μL),phosphate buffer solution(PBS solution 125μL)group and miR-125b-5p simulator(agomiR-125b-5b group,agomiR-125b-5p dry powder 33μg+PBS solution 125μL)was injected intravenously once a week for 2 months.After the intervention,the tibia of the rats in each group was scanned by micro-CT.Bone volume fraction(BV/TV),bone density(BMD)trabecular number(Tb.N),trabecular separation(Tb.Sp)and structural pattern index(SMI)were collected.Target gene prediction,gene ontology(GO)function and Kyoto Gene and Genome Encyclopedia(KEGG)signaling pathway enrichment analysis of miR-125b-5p were performed using bioinformatics techniques.Results Compared with the healthy people,there were 956 different mirnas in the plasma of OP patients,including 502 up-regulated and 454 down-regulated mirnas,especially the increased expression of miR-125b-5p.The BV/TV and BMD of tibia in OP group were lower than those in Sham group(P<0.05 or P<0.01),suggesting that the osteoporosis model was successfully constructed.Compared with agomiR-NC group,BV/TV and Tb.N of tibia in agomiR-125b-5p group decreased,while Tb.Sp and SMI increased(P<0.05),but there was no statistical signifi
作者
王槐骏
杨城
李玮民
冉栋成
彭维艳
王春庆
WANG Huaijun;YANG Cheng;LI Weimin;RAN Dongcheng;PENG Weiyan;WANG Chunqing(School of Clinical Medicine,Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Emergency Surgery,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China)
出处
《贵州医科大学学报》
CAS
2023年第11期1292-1299,1306,共9页
Journal of Guizhou Medical University
基金
国家自然科学基金(NS20042)
贵州医科大学国家自然科学基金培育项目(学术新苗)(19NSP046)。
