间充质干细胞衍生的外泌体 microRNA-136-5p 通过靶向 ELF3 抑制创伤性骨关节炎中的软骨细胞退变。
Mesenchymal stem cell-derived exosomal microRNA-136-5p inhibits chondrocyte degeneration in traumatic osteoarthritis by targeting ELF3.
机构信息
Department of Orthopedics, The Second Hospital of Jilin University, No. 218, Ziqiang Street, Nanguan District, Changchun, 130041, Jilin Province, People's Republic of China.
Department of Nursing, The Second Hospital of Jilin University, Changchun, 130041, People's Republic of China.
出版信息
Arthritis Res Ther. 2020 Oct 27;22(1):256. doi: 10.1186/s13075-020-02325-6.
BACKGROUND
Emerging evidence suggests that microRNAs (miRs) are associated with the progression of osteoarthritis (OA). In this study, the role of exosomal miR-136-5p derived from mesenchymal stem cells (MSCs) in OA progression is investigated and the potential therapeutic mechanism explored.
METHODS
Bone marrow mesenchymal stem cells (BMMSCs) and their exosomes were isolated from patients and identified. The endocytosis of chondrocytes and the effects of exosome miR-136-5p on cartilage degradation were observed and examined by immunofluorescence and cartilage staining. Then, the targeting relationship between miR-136-5p and E74-like factor 3 (ELF3) was analyzed by dual-luciferase report assay. Based on gain- or loss-of-function experiments, the effects of exosomes and exosomal miR-136-5p on chondrocyte migration were examined by EdU and Transwell assay. Finally, a mouse model of post-traumatic OA was developed to evaluate effects of miR-136-5p on chondrocyte degeneration in vivo.
RESULTS
In the clinical samples of traumatic OA cartilage tissues, we detected increased ELF3 expression, and reduced miR-136-5p expression was determined. The BMMSC-derived exosomes showed an enriched level of miR-136-5p, which could be internalized by chondrocytes. The migration of chondrocyte was promoted by miR-136-5p, while collagen II, aggrecan, and SOX9 expression was increased and MMP-13 expression was reduced. miR-136-5p was verified to target ELF3 and could downregulate its expression. Moreover, the expression of ELF3 was reduced in chondrocytes after internalization of exosomes. In the mouse model of post-traumatic OA, exosomal miR-136-5p was found to reduce the degeneration of cartilage extracellular matrix.
CONCLUSION
These data provide evidence that BMMSC-derived exosomal miR-136-5p could promote chondrocyte migration in vitro and inhibit cartilage degeneration in vivo, thereby inhibiting OA pathology, which highlighted the transfer of exosomal miR-136-5p as a promising therapeutic strategy for patients with OA.
背景
新出现的证据表明，微小 RNA（miRs）与骨关节炎（OA）的进展有关。在这项研究中，研究了间充质干细胞（MSCs）衍生的外泌体 miR-136-5p 在 OA 进展中的作用，并探讨了其潜在的治疗机制。
方法
从患者中分离出骨髓间充质干细胞（BMMSCs）及其外泌体并进行鉴定。通过免疫荧光和软骨染色观察和检查软骨细胞的内吞作用以及外泌体 miR-136-5p 对软骨降解的影响。然后，通过双荧光素酶报告实验分析 miR-136-5p 与 E74 样因子 3（ELF3）的靶向关系。基于增益或功能丧失实验，通过 EdU 和 Transwell 测定检查外泌体和外泌体 miR-136-5p 对软骨细胞迁移的影响。最后，建立创伤后 OA 小鼠模型，以评估 miR-136-5p 在体内对软骨细胞退变的影响。
结果
在创伤性 OA 软骨组织的临床样本中，我们检测到 ELF3 表达增加，miR-136-5p 表达减少。BMMSC 来源的外泌体显示 miR-136-5p 水平丰富，可被软骨细胞内化。miR-136-5p 促进了软骨细胞的迁移，同时增加了胶原 II、聚集蛋白聚糖和 SOX9 的表达，减少了 MMP-13 的表达。miR-136-5p 被证实靶向 ELF3 并能下调其表达。此外，外泌体内化后软骨细胞中 ELF3 的表达减少。在创伤后 OA 小鼠模型中，发现外泌体 miR-136-5p 可减少软骨细胞外基质的退变。
结论
这些数据提供了证据表明，BMMSC 衍生的外泌体 miR-136-5p 可促进体外软骨细胞迁移，并抑制体内软骨退变，从而抑制 OA 病理，突出了外泌体 miR-136-5p 作为 OA 患者有前途的治疗策略的转移。
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