激素对大鼠股骨头骨微血管内皮细胞miRNA表达谱的生物信息学探索  

Bioinformatics study on the effect of glucocorticoid on miRNA expression in bone microvascular endothelial cells of femoral head in rats

作　　者：岳聚安 王冉东[1] 张启栋[2] 李兵 孙强[1] 刘忘言 陈蛟 李迎楠 郭晓忠 Yue Ju′an;Wang Randong;Zhang Qidong;Li Bing;Sun Qiang;Liu Wangyan;Chen Jiao;Li Yingnan;Guo Xiaozhong(Department of Orthopedic Surgery, Aviation General Hospital of China Medical University, Beijing 100012, China;Department of Orthopedic Surgery, China- Japan Friendship Hospital, Beijing 100029, China)

机构地区：[1]中国医科大学航空总医院关节外科,北京100012 [2]中日友好医院关节外科,北京100029

出　　处：《中华损伤与修复杂志（电子版）》2020年第2期103-109,共7页Chinese Journal of Injury Repair and Wound Healing(Electronic Edition)

基　　金：国家自然科学基金面上项目(81705896)。

摘　　要：目的探索激素对股骨头骨微血管内皮细胞(BMEC)miRNA表达谱的影响及相关的生物信息学分析。方法选取SPF级8周龄雌性SD大鼠14只,适应性饲养1周后,按照随机数字表法分为2组,模型组7只、空白组7只。模型组腹腔注射脂多糖(20μg/kg),每次间隔24 h,连续注射2次,24 h后肌内注射大剂量甲强龙(40 mg/kg),每次间隔24 h,连续注射3次。空白组与模型组相同的时间点给予等量0.9%氯化钠溶液腹腔注射、肌内注射。注射后4周随机从2组中各取2只大鼠行股骨头病理苏木精-伊红染色,评估造模是否成功,同时行股骨头BMEC培养,形态学观察与鉴定及生物信息学检测如使用Agilent GeneSpring软件对差异表达的miRNA进行筛选,采用miRWalk 2.0软件对差异表达的miRNA进行靶基因预测,Panther数据库被用来预测靶基因的信号通路,Gene Ontology(GO)对靶基因进行功能注释分析。结果通过联合采用脂多糖及甲强龙成功地建立早期股骨头坏死(ONFH)大鼠模型;分离的骨BMEC纯度高达98.5%;与空白组比较,模型组共检测到4个差异有统计学意义异常表达的miRNA(2个上调:miR-132-3p、miR-335,2个下调:miR-466b-2-3p、let-7c-1-3p),其靶基因共3808个;靶基因调控的信号通路可能包括JAK/STAT signaling pathway等,其富集的生物途径包括metabolic process等,富集在细胞定位中包括organelle、nucleoid等,富集在生物学功能中的包括nucleic acid binding transcription factor activity等。结论激素可诱导股骨头BMEC miRNA的表达谱发生变化。miR-132-3p和miR-335的异常表达可能在激素性股骨头坏死发生中起重要作用。Objective To explore the effect of glucocorticoid on miRNA expression profile of femoral head bone microvascular endothelial cell(BMEC)and bioinformatics analysis.Methods Fourteen SPF 8-week-old female SD rats were selected.After 1 week of adaptive feeding,they were divided into 2 groups by random number table method,7 in model group and 7 in control group.The model group was intraperitoneally injected with lipopolysaccharide(20μg/kg)for two consecutive injections 24 h apart,and intramuscular injection of high-dose methylprednisolone(40 mg/kg)for three consecutive injections 24 h apart.The control group was given intraperitoneal injection and intramuscular injection of the same amount of normal saline at the same time.After 4 weeks,2 of the rats from each group were sectioned randomly to confirm the establishment of the model.And the femoral head BMEC were cultured,identified and tested using bioinformatics methods.Such as the Agilent GeneSpring were used to identify the differentially expressed miRNA,miRWalk2.0 software were used to explore target genes,Panther database was used to predict signaling pathway of target genes,Gene ontology(GO)was used to analysis the functional annotation of target genes.Results The rat model of early osteonecrosis of the femoral head(ONFH)was successfully established by using lipopolysaccharide and methylamydron.The purity of BMEC was as high as 98.5%.Compared with control group,4 miRNA(2 up-regulated miRNA:miRNA-132-3p,miRNA-335,and 2 down-regulated miRNA:miRNA-466b-2-3p,let-7c-1-3p)were detected in the model group,3808 target genes of which were detected.The signaling pathways regulated by target genes may include JAK/STAT signaling pathway and so on,their enrichment biological pathways include metabolic process and so on,localization in cell include organelle,nucleoid and so on,enrichment includes this in biological function include nucleic acid binding transcription factor activity and so on.Conclusions Glucocorticoid can induce the changes of miRNA expression profile in femo

关 键 词：激素类 股骨头坏死 内皮细胞 生物信息 

分 类 号：R73[医药卫生—肿瘤]