Department of Cardiac Surgery, The First Affiliated Hospital of Xinjiang Medical University, 137 Liyushan South Road, Xinshi District, Urumqi, 830054, Xinjiang, China.
Department of Cardiology, The First Affiliated Hospital of Xinjiang Medical University, 137 Liyushan South Road, Xinshi District, Urumqi, 830054, Xinjiang, China.
BMC Cardiovasc Disord. 2022 Feb 2;22(1):24. doi: 10.1186/s12872-022-02471-6.
Human aortic valve interstitial cells (hAVICs) are a key factor in the pathogenesis of calcific aortic valve disease (CAVD). This research examines the role and mechanism of microRNA miR-138-5p in osteogenic differentiation of hAVICs.
RT-qPCR analysis was applied for detecting miR-138-5p and RUNX2 expression in valve tissues of CAVD patients and controls. On completion of induction of osteogenic differentiation of hAVICs, and after overexpression or interference of miR-138-5p expression, the condition of osteogenic differentiation and calcification of hAVICs was confirmed using alkaline phosphatase staining and alizarin red staining. Subsequently, western blot was utilized to detect the expression of osteogenesis-related proteins OPN and ALP, and Wnt/β-catenin signaling pathway-related proteins. Finally, the relationship between miR-138-5p and RUNX2 was validated by dual-luciferase reporter assay and Pearson's correlation test.
Down-regulation of miR-138-5p was found in CAVD patients and during osteogenic differentiation of hAVICs. Overexpression of miR-138-5p contribute to the inhibition of osteoblast differentiation and calcium deposition in hAVICs, and of ALP and OPN protein expression. RUNX2 was a target gene of miR-138-5p, and it was negatively correlated with miR-138-5p in CAVD. Additionally, overexpression of RUNX2 could reverse the inhibitory effect of miR-138-5p on osteogenic differentiation of hAVICs.
miR-138-5p can act as a positive regulator of osteogenic differentiation in CAVD patients to involve in inhibiting valve calcification, which is achieved through RUNX2 and Wnt/β-catenin signaling pathway.
人主动脉瓣间质细胞（hAVICs）是钙化性主动脉瓣病（CAVD）发病机制中的关键因素。本研究探讨了 microRNA miR-138-5p 在 hAVICs 成骨分化中的作用和机制。
应用 RT-qPCR 分析检测 CAVD 患者和对照者瓣膜组织中 miR-138-5p 和 RUNX2 的表达。在 hAVICs 成骨诱导分化完成后，以及过表达或干扰 miR-138-5p 表达后，通过碱性磷酸酶染色和茜素红染色确认 hAVICs 的成骨分化和钙化情况。随后，采用 Western blot 检测成骨相关蛋白 OPN 和 ALP 的表达，以及 Wnt/β-catenin 信号通路相关蛋白的表达。最后，通过双荧光素酶报告基因检测和 Pearson 相关检验验证 miR-138-5p 与 RUNX2 的关系。
CAVD 患者和 hAVICs 成骨分化过程中 miR-138-5p 表达下调。过表达 miR-138-5p 可抑制 hAVICs 成骨分化和钙沉积，降低 ALP 和 OPN 蛋白表达。RUNX2 是 miR-138-5p 的靶基因，在 CAVD 中与 miR-138-5p 呈负相关。此外，过表达 RUNX2 可逆转 miR-138-5p 对 hAVICs 成骨分化的抑制作用。
miR-138-5p 可作为 CAVD 患者成骨分化的正调节剂，通过 RUNX2 和 Wnt/β-catenin 信号通路参与抑制瓣膜钙化。
