Department of Vascular Surgery, Suzhou TCM Hospital Affiliated to Nanjing University of Chinese Medicine, Suzhou 215000, China.
Department of Orthopedics, The Second Affiliated Hospital of Soochow University, Suzhou 215006, China.
Exp Biol Med (Maywood). 2021 Nov;246(21):2259-2268. doi: 10.1177/15353702211023034. Epub 2021 Jun 30.
Deep venous thrombosis is one of the most common venous thromboembolic diseases and has a low cure rate and a high postoperative recurrence rate. Furthermore, emerging evidence indicates that microRNAs are involved in deep venous thrombosis. miR-296-5p is an important microRNA that plays a critical role in various cellular functions, and S100A4 is closely related to vascular function. miR-296-5p is downregulated in deep venous thrombosis patients, and its predicted target S100A4 is upregulated in deep venous thrombosis patients. Therefore, it was hypothesized that miR-296-5p may play a vital role in the development of deep venous thrombosis by targeting S100A4. An Ox-LDL-stimulated HUVEC and deep venous thrombosis mouse model was employed to detect the biological functions of miR-296-5p and S100A4. Dual luciferase reporter assays and pull-down assays were used to authenticate the interaction between miR-296-5p and S100A4. ELISA and Western blotting were employed to detect the protein levels of thrombosis-related factors and the endothelial-to-mesenchymal transition (EndMT)-related factors. The miR-296-5p levels were reduced, while the S100A4 levels were enhanced in deep venous thrombosis patients, and the miR-296-5p levels were negatively correlated with the S100A4 levels in deep venous thrombosis patients. miR-296-5p suppressed S100A4 expression by targeting the 3' UTR of S100A4. MiR-296-5p knockdown accelerated ox-LDL-induced HUVEC apoptosis, oxidative stress, thrombosis-related factor expression, and EndMT, while S100A4 knockdown antagonized these effects in ox-LDL-induced HUVECs. S100A4 knockdown reversed the effect induced by miR-296-5p knockdown. Moreover, the studies revealed that miR-296-5p knockdown in deep venous thrombosis mice exacerbated deep venous thrombosis formation, whereas S100A4 knockdown had the opposite effect. These results indicate that elevated miR-296-5p inhibits deep venous thrombosis formation by inhibiting S100A4 expression. Both miR-296-5p and S100A4 may be potential diagnostic markers and therapeutic targets for deep venous thrombosis.
深静脉血栓形成是最常见的静脉血栓栓塞疾病之一，其治愈率低，术后复发率高。此外，新出现的证据表明 microRNAs 参与深静脉血栓形成。miR-296-5p 是一种重要的 microRNA，在各种细胞功能中发挥关键作用，而 S100A4 与血管功能密切相关。深静脉血栓形成患者中 miR-296-5p 下调，其预测靶标 S100A4 在深静脉血栓形成患者中上调。因此，推测 miR-296-5p 可能通过靶向 S100A4 在深静脉血栓形成的发展中发挥重要作用。采用 Ox-LDL 刺激的 HUVEC 和深静脉血栓形成小鼠模型来检测 miR-296-5p 和 S100A4 的生物学功能。双荧光素酶报告基因检测和 pull-down 检测用于验证 miR-296-5p 和 S100A4 之间的相互作用。ELISA 和 Western blotting 用于检测血栓形成相关因子和内皮-间质转化 (EndMT) 相关因子的蛋白水平。深静脉血栓形成患者中 miR-296-5p 水平降低，而 S100A4 水平升高，深静脉血栓形成患者中 miR-296-5p 水平与 S100A4 水平呈负相关。miR-296-5p 通过靶向 S100A4 的 3'UTR 抑制 S100A4 表达。miR-296-5p 敲低加速 ox-LDL 诱导的 HUVEC 凋亡、氧化应激、血栓形成相关因子表达和 EndMT，而 S100A4 敲低拮抗 ox-LDL 诱导的 HUVEC 中的这些作用。S100A4 敲低逆转了 miR-296-5p 敲低引起的作用。此外，研究表明深静脉血栓形成小鼠中 miR-296-5p 敲低加剧深静脉血栓形成，而 S100A4 敲低则有相反的作用。这些结果表明，升高的 miR-296-5p 通过抑制 S100A4 表达抑制深静脉血栓形成。miR-296-5p 和 S100A4 均可能成为深静脉血栓形成的潜在诊断标志物和治疗靶点。
