Exosomes from IL-1β stimulated synovial fibroblasts induce osteoarthritic changes in articular chondrocytes.
Exosomes from IL-1β stimulated synovial fibroblasts induce osteoarthritic changes in articular chondrocytes.
复制标题
来自IL-1β的外泌体刺激滑膜成纤维细胞诱导关节软骨细胞的骨关节炎变化。
DOI：
10.1186/ar4679
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发表时间：
2014-08-04
影响因子：
4.9
通讯作者：
Ochi M
中科院分区：
文献类型：
--
作者：
Kato T;Miyaki S;Ishitobi H;Nakamura Y;Nakasa T;Lotz MK;Ochi M
骨关节炎（OA）是一种累及整个关节的疾病，其特征是关节软骨进行性退变、滑膜增生、骨重塑以及各种关节组织中的血管生成。外泌体是一种微泡（MVs），可能在稳态和疾病状态下的组织 - 组织和细胞 - 细胞通讯中发挥作用。我们假设外泌体在一个新的调节网络中发挥作用，该网络有助于骨关节炎的发病机制，并研究了外泌体在关节组织细胞间通讯中的功能。
从正常膝关节获取人滑膜成纤维细胞（SFB）和关节软骨细胞。对从SFB条件培养基中分离出的外泌体进行大小、数量、标志物和功能分析。用来自SFB的外泌体处理正常关节软骨细胞，并用白细胞介素 - 1β（IL - 1β）刺激SFB。使用实时定量PCR对骨关节炎相关基因的表达进行定量。为了分析外泌体对软骨组织的影响，我们进行了糖胺聚糖释放试验。在迁移和管形成试验中测试这些外泌体的血管生成活性。通过Bio - Plex多重检测和NanoString分析对外泌体中的细胞因子和微小RNA（miRNAs）进行分析。
与源自SFB的外泌体相比，来自白细胞介素 - 1β刺激的SFB的外泌体显著上调关节软骨细胞中基质金属蛋白酶 - 13（MMP - 13）和含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶 - 5（ADAMTS - 5）的表达，并下调Ⅱ型胶原蛋白α1链（COL2A1）和聚集蛋白聚糖（ACAN）的表达。用人白细胞介素 - 1β刺激的SFB的外泌体处理的人脐静脉内皮细胞（HUVECs）的迁移和管形成活性显著更高，其还显著诱导软骨外植体释放更多的蛋白聚糖。外泌体中的炎症细胞因子白细胞介素 - 6（IL - 6）、基质金属蛋白酶 - 3（MMP - 3）和血管内皮生长因子（VEGF）仅能在低水平检测到。白细胞介素 - 1β、肿瘤坏死因子α（TNFα）、基质金属蛋白酶 - 9（MMP - 9）和基质金属蛋白酶 - 13（MMP - 13）在外泌体中无法检测到。NanoString分析显示，与未受刺激的SFB相比，来自白细胞介素 - 1β刺激的SFB的外泌体中50种微小RNA的水平存在差异表达。
来自白细胞介素 - 1β刺激的SFB的外泌体在体外和离体模型中均诱导类似骨关节炎的变化。外泌体代表了一种新的机制，通过该机制致病信号在受骨关节炎影响的关节中的不同细胞类型之间传递。
Osteoarthritis (OA) is a whole joint disease, and characterized by progressive degradation of articular cartilage, synovial hyperplasia, bone remodeling and angiogenesis in various joint tissues. Exosomes are a type of microvesicles (MVs) that may play a role in tissue-tissue and cell-cell communication in homeostasis and diseases. We hypothesized that exosomes function in a novel regulatory network that contributes to OA pathogenesis and examined the function of exosomes in communication among joint tissue cells. Human synovial fibroblasts (SFB) and articular chondrocytes were obtained from normal knee joints. Exosomes isolated from conditioned medium of SFB were analyzed for size, numbers, markers and function. Normal articular chondrocytes were treated with exosomes from SFB, and Interleukin-1β (IL-1β) stimulated SFB. OA-related genes expression was quantified using real-time PCR. To analyze exosome effects on cartilage tissue, we performed glycosaminoglycan release assay. Angiogenic activity of these exosomes was tested in migration and tube formation assays. Cytokines and miRNAs in exosomes were analyzed by Bio-Plex multiplex assay and NanoString analysis. Exosomes from IL-1β stimulated SFB significantly up-regulated MMP-13 and ADAMTS-5 expression in articular chondrocytes, and down-regulated COL2A1 and ACAN compared with SFB derived exosomes. Migration and tube formation activity were significantly higher in human umbilical vein endothelial cells (HUVECs) treated with the exosomes from IL-1β stimulated SFB, which also induced significantly more proteoglycan release from cartilage explants. Inflammatory cytokines, IL-6, MMP-3 and VEGF in exosomes were only detectable at low level. IL-1β, TNFα MMP-9 and MMP-13 were not detectable in exosomes. NanoString analysis showed that levels of 50 miRNAs were differentially expressed in exosomes from IL-1β stimulated SFB compared to non-stimulated SFB. Exosomes from IL-1β stimulated SFB induce OA-like changes both in vitro and in ex vivo models. Exosomes represent a novel mechanism by which pathogenic signals are communicated among different cell types in OA-affected joints.
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DOI：
10.1186/1758-907x-1-7
发表时间：
2010-03-01
期刊：
Silence
影响因子：
--
作者：
Kosaka N;Izumi H;Sekine K;Ochiya T
通讯作者：
Ochiya T
影响因子：
21.3
作者：
Hergenreider, Eduard;Heydt, Susanne;Dimmeler, Stefanie
通讯作者：
Dimmeler, Stefanie
影响因子：
7
作者：
Kirsch, T;Swoboda, B;Nah, HD
通讯作者：
Nah, HD
骨关节炎的炎症。
影响因子：
5.1
作者：
Goldring MB;Otero M
通讯作者：
Otero M
影响因子：
3.8
作者：
King HW;Michael MZ;Gleadle JM
通讯作者：
Gleadle JM