关节滑膜液Exosomes激活YAP蛋白保护关节软骨的机制研究  

Mechanism of Synovial Fluid Exosomes to Protect Articular Cartilage Through Activating YAP Protein

作　　者：高坤 朱文秀[2] 李亨[1] 刘伟东[1] 马笃军[1] 余伟吉[1] 曹亚飞[1] Gao Kun;Zhu Wenxiu;Li Heng;Liu Weidong;Ma Dujun;Yu Weiji;Cao Yafei(Shenzhen Traditional Chinese Medicine Hospital,Shenzhen,518100;Shenzhen Hospital of Beijing University of Traditional Chinese Medicine,Shenzhen,518100)

机构地区：[1]深圳市中医院,深圳518000 [2]北京中医药大学深圳医院,深圳518000

出　　处：《基因组学与应用生物学》2019年第8期3781-3785,共5页Genomics and Applied Biology

摘　　要：为了研究膝关节炎滑膜液中外泌体(Exosomes)对软骨的作用,并探讨其通过何种信号通路发挥作用,本研究通过ExoQuick提取滑膜液中Exosomes并用电镜、Nanosight、Western blotting进行鉴定;采用四甲基偶氮唑盐比色法(MTT)检测软骨组织细胞活性;酶联免疫吸附试验(ELISA)法检测采用上清液中MMP-1、MMP-13浓度;Western blotting检测蛋白磷酸化水平;实时荧光定量PCR方法检测下游基因表达水平。研究结果显示成功提取大小在100 nm以下Exosomes颗粒。关节炎滑膜液来源Exosomes与空白对照及正常Exosomes相比较,能够明显促进软骨细胞的增殖,降低MMP-1、MMP-13水平。检测YAP蛋白发生明显磷酸化,与对照组比较有明显差异,YAP下游基因CTGF、Cyr61、ANKRD1均明显高表达。膝关节炎滑膜液分泌的Exosomes能够通过YAP的去磷酸化,抑制MMP-1、MMP-13的分泌,促进软骨细胞的增殖,具有保护关节软骨的作用。To study the effect of exosomes derived from synovial fluid of knee joint on cartilage and explore the signaling pathway.Exosomes were extracted from synovial fluid by Exo Quick and identified by electron microscopy,nanosight and Western blotting.Cartilage cells were assayed for cell viability by MTT assay.Enzyme-linked immunosorbent assay(ELISA)was used to detect the concentration of MMP-1 and MMP-13 in the supernatant.Western blot was used to detect the level of protein phosphorylation,and real-time fluorescence quantitative PCR was used to detect the level of downstream gene expression.The results showed that the exosomes particles below 100 nm were successfully extracted.Arthritis synovial fluid exosomes can significantly promote the proliferation of chondrocytes and reduce MMP-1,MMP-13 levels,compared with the blank control and normal.

关 键 词：关节炎 外泌体 YAP蛋白 关节软骨 保护机制 

分 类 号：R73[医药卫生—肿瘤]