PIEZOl/miR-155-5p/GDF6/SMAD2/3 信号轴参与了在过度机械应力下诱导骨关节炎的发生和进展。

The PIEZO1/miR-155-5p/GDF6/SMAD2/3 signaling axis is involved in inducing the occurrence and progression of osteoarthritis under excessive mechanical stress.

机构信息

Nanjing First Hospital, Nanjing Medical University, China.

Nanjing First Hospital, Nanjing Medical University, China..

出版信息

Cell Signal. 2024 Jun;118:111142. doi: 10.1016/j.cellsig.2024.111142. Epub 2024 Mar 18.

DOI:10.1016/j.cellsig.2024.111142

PMID:38508350

Abstract

OBJECTIVE

To elucidate the molecular mechanism of overloading-induced osteoarthritis (OA) and to find a novel therapeutic target.

METHODS

We utilized human cartilage specimens, mouse chondrocytes, a destabilization of the medial meniscus (DMM) mouse model, and a mouse hindlimb weight-bearing model to validate the role of overloading on chondrocyte senescence and OA development. Then, we observed the effect of PIEZO1-miR-155-5p-GDF6-SMAD2/3 signaling axis on the preservation of joint metabolic homeostasis under overloading in vivo, in vitro and ex vivo by qPCR, Western blot, enzyme-linked immunosorbent assay (ELISA), immunohistochemistry, immunofluorescence, SA-β-gal staining, CCK8 assay, et al. Finally, we verified the therapeutic effects of intra-articular injection of miR-155-5p inhibitor or recombinant GDF6 on the murine overloading-induced OA models.

RESULTS

Chondrocytes sensesed the mechanical overloading through PIEZO1 and up-regulated miR-155-5p expression. MiR-155-5p mimics could copy the effects of overloading-induced chondrocyte senescence and OA. Additionally, miR-155-5p could suppress the mRNA expression of Gdf6-Smad2/3 in various tissues within the joint. Overloading could disrupt joint metabolic homeostasis by downregulating the expression of anabolism indicators and upregulating the expression of catabolism indicators in the chondrocytes and synoviocytes, while miR-155-5p inhibition or GDF6 supplementation could exert an antagonistic effect by preserving the joint homeostasis. Finally, in the in vivo overloading models, intra-articular injection of miR-155-5p inhibitor or recombinant GDF6 could significantly mitigate the severity of impending OA and lessened the progression of existing OA.

CONCLUSION

GDF6 overexpression or miR-155-5p inhibition could attenuate overloading-induced chondrocyte senescence and OA through the PIEZO1-miR-155-5p-GDF6-SMAD2/3 signaling pathway. Our study provides a new therapeutic target for the treatment of overloading-induced OA.

摘要

目的

阐明超负荷诱导骨关节炎（OA）的分子机制，寻找新的治疗靶点。

方法

我们利用人软骨标本、小鼠软骨细胞、内侧半月板不稳定（DMM）小鼠模型和小鼠后肢负重模型来验证超负荷对软骨细胞衰老和 OA 发展的作用。然后，我们通过 qPCR、Western blot、酶联免疫吸附试验（ELISA）、免疫组织化学、免疫荧光、SA-β-半乳糖苷染色、CCK8 测定等方法，观察 PIEZO1-miR-155-5p-GDF6-SMAD2/3 信号轴在体内、体外和离体超负荷状态下对关节代谢稳态的维持作用。最后，我们验证了关节内注射 miR-155-5p 抑制剂或重组 GDF6 对小鼠超负荷诱导 OA 模型的治疗效果。

结果

软骨细胞通过 PIEZO1 感知机械超负荷，并上调 miR-155-5p 表达。miR-155-5p 模拟物可以复制超负荷诱导的软骨细胞衰老和 OA 效应。此外，miR-155-5p 可以抑制关节内各种组织中 Gdf6-Smad2/3 的 mRNA 表达。超负荷通过下调软骨细胞和滑膜细胞中合成代谢标志物的表达，上调分解代谢标志物的表达，破坏关节代谢稳态，而 miR-155-5p 抑制或 GDF6 补充可以通过维持关节稳态发挥拮抗作用。最后，在体内超负荷模型中，关节内注射 miR-155-5p 抑制剂或重组 GDF6 可以显著减轻即将发生的 OA 的严重程度，并减缓现有的 OA 的进展。