Zhang Yunhong, Zhang Zhen, Li Haoyang, Chu Chu, Liang Gang, Fan Nannan, Wei Ran, Zhang Tingting, Li Lihua, Wang Bin, Li Xia
Innovative Institute of Chinese Medicine and Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan, Shandong Province, China.
International Business School, Tianjin Foreign Studies University, Tianjin, China.
Front Cardiovasc Med. 2024 Mar 20;11:1356286. doi: 10.3389/fcvm.2024.1356286. eCollection 2024.
Deep vein thrombosis (DVT) is associated with aberrant gene expression that is a common peripheral vascular disease. Here, we aimed to elucidate that the epigenetic modification of forkhead box protein 3 (FOXP3) at the post-transcriptional level, which might be the key trigger leading to the down-regulation of FOXP3 expression in DVT.
In order to explore the relationship between microRNAs (miRNAs) and FOXP3, mRNA and microRNA microarray analysis were performed. Dual luciferase reporter assay was used to verify the upstream miRNAs of FOXP3. Quantitative real-time polymerase chain reaction, flow cytometry and Western blot were used to detect the relative expression of miR-6132 and FOXP3. Additionally, DVT models were established to investigate the role of miR-6132 by Murine Doppler Ultrasound and Hematoxylin-Eosin staining.
Microarray and flow cytometry results showed that the FOXP3 expression was decreased while miR-6132 level was increased substantially in DVT, and there was significant negative correlation between miR-6132 and FOXP3. Moreover, we discovered that overexpressed miR-6132 reduced FOXP3 expression and aggravated DVT formation, while miR-6132 knockdown increased FOXP3 expression and alleviated DVT formation. Dual luciferase reporter assay validated the direct binding of miR-6132 to FOXP3.
Collectively, our data elucidate a new avenue through which up-regulated miR-6132 contributes to the formation and progression of DVT by inhibiting FOXP3 expression.
深静脉血栓形成（DVT）与异常基因表达相关，是一种常见的周围血管疾病。在此，我们旨在阐明转录后水平上叉头框蛋白3（FOXP3）的表观遗传修饰，这可能是导致DVT中FOXP3表达下调的关键触发因素。
为了探究微小RNA（miRNA）与FOXP3之间的关系，进行了mRNA和miRNA微阵列分析。采用双荧光素酶报告基因检测法验证FOXP3的上游miRNA。运用定量实时聚合酶链反应、流式细胞术和蛋白质免疫印迹法检测miR-6132和FOXP3的相对表达。此外，建立DVT模型，通过小鼠多普勒超声和苏木精-伊红染色研究miR-6132的作用。
微阵列和流式细胞术结果显示，DVT中FOXP3表达降低，而miR-6132水平显著升高，且miR-6132与FOXP3之间存在显著负相关。此外，我们发现过表达miR-6132会降低FOXP3表达并加重DVT形成，而敲低miR-6132则会增加FOXP3表达并减轻DVT形成。双荧光素酶报告基因检测法验证了miR-6132与FOXP3的直接结合。
总体而言，我们的数据阐明了一条新途径，即上调的miR-6132通过抑制FOXP3表达促进DVT的形成和进展。
