miR-137 通过靶向抑制 TCF4 逆转 AMPK/NF-κB 信号通路促进骨关节炎进展。

miR-137 targets the inhibition of TCF4 to reverse the progression of osteoarthritis through the AMPK/NF-κB signaling pathway.

机构信息

Department of Spinal and Joint Surgery, Qingdao Chengyang District People's Hospital, Qingdao City 266109, Shandong Province, China.

Department of Radiology, Qingdao Chengyang District People's Hospital, Qingdao City 266109, Shandong Province, China.

出版信息

Biosci Rep. 2020 Jun 26;40(6). doi: 10.1042/BSR20200466.

PURPOSE

To explore the regulatory mechanism of miR-137 and transcription factor 4 (TCF4) in the progression of osteoarthritis (OA).

PATIENTS AND METHODS

The expressions of miR-137 and TCF4 were detected in OA cartilage tissue, chondrocytes and OA rat cartilage tissue. miR-137 and TCF4 were up-regulated or down-regulated and transfected into chondrocytes and OA rat cartilage tissue. The gene expression, protein level, cell proliferation, apoptosis and inflammatory factors were detected, respectively. LPS and anterior cruciate ligament transection (ACLT) on the right knee were used to induce chondrocyte inflammation and establish rat OA model, respectively.

RESULTS

miR-137 was low expressed in cartilage tissue of OA group, while TCF4 expression and protein level were significantly higher, showing significant negative correlation. In LPS group, chondrocyte activity was significantly inhibited, cell apoptosis ability was significantly enhanced, and the levels of inflammatory factors TNF-α, IL-1β, IL-6 were significantly increased. However, the above results were significantly improved after the up-regulation of miR-137 or down-regulation of TCF4. Double luciferase report revealed that miR-137 and TCF4 had targeted relationship. LPS induced activation of AMPK/NF-κB pathway and higher level of apoptosis. AMPK/NF-κB pathway inhibitor C could inhibit activation of this pathway, and up-regulation of miR-137 or down-regulation of TCF4 could significantly weaken the regulation of LPS on the pathway and apoptosis. Analysis of OA rat model showed that over-expression of miR-137 could inhibit up-regulation of inflammatory factors and activation of AMPK/NF-κB pathway.

CONCLUSION

miR-137 targets the inhibition of TCF4 to reverse the progression of OA through the AMPK/NF-κB signaling pathway.

目的

探索 microRNA-137（miR-137）和转录因子 4（TCF4）在骨关节炎（OA）进展中的调控机制。

方法

检测 OA 软骨组织、软骨细胞和 OA 大鼠软骨组织中 miR-137 和 TCF4 的表达。上调或下调 miR-137 和 TCF4 并转染至软骨细胞和 OA 大鼠软骨组织。分别检测基因表达、蛋白水平、细胞增殖、凋亡和炎症因子。分别采用 LPS 和前交叉韧带切断（ACLT）术处理右侧膝关节以诱导软骨细胞炎症和建立大鼠 OA 模型。

结果

OA 组软骨组织中 miR-137 表达水平较低，而 TCF4 表达和蛋白水平显著升高，呈显著负相关。在 LPS 组，软骨细胞活性明显受到抑制，细胞凋亡能力明显增强，TNF-α、IL-1β、IL-6 等炎症因子水平显著升高。然而，上调 miR-137 或下调 TCF4 后，上述结果均明显改善。双荧光素酶报告显示，miR-137 和 TCF4 具有靶向关系。LPS 诱导 AMPK/NF-κB 通路激活和凋亡水平升高。AMPK/NF-κB 通路抑制剂 C 可抑制该通路的激活，上调 miR-137 或下调 TCF4 可显著减弱 LPS 对该通路和凋亡的调节作用。OA 大鼠模型分析表明，miR-137 的过表达可抑制炎症因子的上调和 AMPK/NF-κB 通路的激活。