微小 RNA-185 的过表达通过抑制 Wnt/-连环蛋白信号通路和下调半乳糖凝集素-3 来减轻椎间盘退变。
Overexpression of microRNA-185 alleviates intervertebral disc degeneration through inactivation of the Wnt/-catenin signaling pathway and downregulation of Galectin-3.
机构信息
Department of Colorectal and Anal Surgery, The First Hospital of Jilin University, Changchun, P. R. China.
Day Care Unit, The First Hospital of Jilin University, Changchun, P. R. China.
出版信息
Mol Pain. 2020 Jan-Dec;16:1744806920902559. doi: 10.1177/1744806920902559.
BACKGROUND
Recent studies have found that microRNAs (miRNAs) play a critical role in development and progression of intervertebral disc degeneration. In the present study, we examined the role of miR-185 in nucleus pulposus cell behavior in vitro and the histological changes of intervertebral disc tissue in intervertebral disc degeneration rat models in vivo.
METHODS
Intervertebral disc degeneration models were developed in Sprague-Dawley rats. Intervertebral disc tissue was collected for histological evaluation after miR-185 agomir/agomir transduction. Next, nucleus pulposus tissues were collected from lumbar intervertebral discs to isolate nucleus pulposus cells, which were treated with miR-185 mimic/inhibitor and an inhibitor of the Wnt signaling pathway to assess cell viability and apoptosis.
RESULTS
We observed a high expression of Galectin-3 in nucleus pulposus cells of rats with intervertebral disc degeneration. Bioinformatics prediction and dual-luciferase reporter assay confirmed that miR-185 specifically binds to and negatively regulates Galectin-3. Furthermore, we found that miR-185 inhibition resulted in increased expression of Galectin-3, pro-autophagy factors (LC3 and Beclin-1), and pro-apoptosis factors (caspase-3 and Bax), along with the activation of the Wnt/β-catenin signaling pathway. Moreover, the gain- and loss-of-function studies suggested that miR-185 overexpression promoted cell viability and inhibited nucleus pulposus cell apoptosis and autophagy inactivation of the Wnt/β-catenin signaling pathway. Moreover, miR-185 agomir alleviated the histological changes observed in intervertebral disc tissues in intervertebral disc degeneration rats, which helped us validate the results observed in vitro.
CONCLUSIONS
Overexpression of miR-185 promotes nucleus pulposus cell viability and reduces the histological changes observed in intervertebral disc tissues in rats with intervertebral disc degeneration inactivation of the Wnt/β-catenin signaling pathway and Galectin-3 inhibition. Our findings also highlight the potential of miR-185 as a promising novel therapeutic target to prevent and control intervertebral disc degeneration.
背景
最近的研究发现 microRNAs（miRNAs）在椎间盘退变的发生和发展中起着关键作用。在本研究中，我们研究了 miR-185 在体外核髓细胞行为以及体内椎间盘退变大鼠模型椎间盘组织组织学变化中的作用。
方法
建立 Sprague-Dawley 大鼠椎间盘退变模型。转染 miR-185 agomir/agomir 后，收集椎间盘组织进行组织学评价。然后，从腰椎椎间盘采集髓核组织，分离髓核细胞，用 miR-185 模拟物/抑制剂和 Wnt 信号通路抑制剂处理，评估细胞活力和细胞凋亡。
结果
我们观察到椎间盘退变大鼠髓核细胞中 Galectin-3 高表达。生物信息学预测和双荧光素酶报告基因实验证实，miR-185 特异性结合并负调控 Galectin-3。此外，我们发现 miR-185 抑制导致 Galectin-3、自噬前体（LC3 和 Beclin-1）和促凋亡因子（caspase-3 和 Bax）表达增加，同时激活 Wnt/β-catenin 信号通路。此外，功能获得和缺失研究表明，miR-185 过表达促进细胞活力，抑制髓核细胞凋亡和自噬，抑制 Wnt/β-catenin 信号通路。此外，miR-185 agomir 减轻了椎间盘退变大鼠椎间盘组织中观察到的组织学变化，这验证了我们在体外观察到的结果。
结论
miR-185 的过表达促进了核髓细胞的活力，减少了椎间盘退变大鼠椎间盘组织中观察到的组织学变化，同时抑制了 Wnt/β-catenin 信号通路和 Galectin-3 的抑制。我们的研究结果还强调了 miR-185 作为一种有前途的新型治疗靶点，预防和控制椎间盘退变的潜力。
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