清络通痹方对佐剂性关节炎大鼠破骨细胞分化相关miRNA表达的影响  

Effect of Qingluo Tongbi Compound on Osteoclast Differentiation-related miRNA Expressions in Adjuvant Induced Arthritis Rats

作　　者：彭孝武[1] 周玲玲[2] 朱亚梅[1] 周学平[1] 

机构地区：[1]南京中医药大学第一临床医学院,南京210023 [2]南京中医药大学药学院,南京210023

出　　处：《中国中西医结合杂志》2016年第10期1213-1218,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家基金预研基金资助项目(No.14XYY01)

摘　　要：目的观察清络通痹方(Qingluo Tongbi Compound,QTC)对佐剂性关节炎(adjuvant induced arthritis,AIA)大鼠骨破坏相关miRNA表达的影响,探讨其治疗类风湿关节炎的机制。方法建立AIA大鼠滑膜成纤维细胞和单核细胞共培养诱生破骨样细胞模型,应用miRNA芯片的方法初步筛选破骨细胞分化后期miRNA的异常表达谱,并采用实时定量PCR(RT-PCR)对芯片结果进行验证。制备QTC含药血清和空白血清,将破骨细胞随机分为空白组、空白血清组及QTC组,采用RT-PCR检测QTC对差异表达miRNA的影响,并应用生物信息学软件对关键性miRNA进行分析。结果与单核细胞比较,破骨样细胞分化过程中存在明显差异表达的miRNAs共211个,其中表达上调88个,表达下调123个。RT-PCR验证结果与芯片检测结果表达趋势一致。RT-PCR结果显示,QTC干预后miR-140-5p表达明显上调,生物信息学分析结果显示miR-140-5p靶基因显著富集在肌动蛋白细胞骨架的调节、Ras信号通路、c AMP信号通路和Rap1信号通路等信号通路上。结论破骨样细胞分化后期存在多种miRNAs失调,QTC可能通过影响miR-140-5p的表达参与调控破骨细胞的分化。Objective To observe the effect of Qingluo Tongbi Compound（ QTC） on osteoclast differentiation-related miRNA expressions in adjuvant induced arthritis（ AIA） rats,and to study its mechanism for treating rheumatoid arthritis（ RA）. Methods The synovial fibroblasts and monocytes of peripheral blood from AIA rats were co-cultured to induce osteoclast-like cells. Differently expressed miRNAs in the late stage osteoclasts differentiation were detected by miRCURY^TM Array. Real-time quantitative PCR（ RT-PCR） was applied to verify the reliability of miRNA array. QTC drug-containing sera and blank sera were prepared and added to the co-cultured system. The osteoclasts were randomly divided into three groups,the blank group,the blank serum group,and the QTC group. RT-PCR was applied to detect the effect of QTC on related differentially expressed miRNAs. Bioinformatics software was applied to analyze related differentially expressed miRNAs. Results miRNA array results showed that as compared with the monocytes group,there were 211 miRNAs differentially expressed in osteoclast-like cell differentiation,including 88up-regulated miRNAs and 123 down-regulated miRNAs. Results of RT-PCR were consistent with results of the array.RT-PCR showed that the expression level of miR-140-5p was obviously up-regulated after the intervention of QTC.Results of bioinformatics analyses showed that the target gene of miR-140-5p was significantly enriched in signaling pathways such as the regulation of actin cytoskeleton,Ras signaling pathways,c AMP signaling pathways,and Rap1 signaling pathways. Conclusions There were various dysregulated expressions of miRNAs in the anaphase of osteo-clast-like cells differentiation. QTC participated the regulation of osteoclast differentiation by effecting the expression of miR-140-5p.

关 键 词：清络通痹方 MICRORNA 类风湿关节炎 破骨细胞 

分 类 号：R285.5[医药卫生—中药学]