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Abstract：
Aortic aneurysm and dissection (AD) represent a critical cardiovascular emergency with an alarmingly high mortality rate. Recent research has spotlighted the overexpression of genes associated with the m6A modification in AD patients, linking them to the presence of inflammatory M1-type macrophages. Moreover, glycolysis is widely recognized as a key feature of inflammatory M1-type macrophages, but biomarkers linking glycolysis and macrophage function to promote disease progression in AD have not been reported. We conducted an analysis of aortic immune cell infiltration, macrophages, and m6A-related biomarkers in AD patients using bioinformatics techniques. Subsequently, we employed a combination of RT-PCR, WB, and immunofluorescence assays to elucidate the alterations in the expression of M1- and M2-type macrophages, as well as markers of glycolysis, following the overexpression of key biomarkers. These findings were further validated in vivo through the creation of a rat model of AD with knockdown of the aforementioned key biomarkers. The findings revealed that the m6A-modified related gene RBM15 exhibited heightened expression in AD samples and was correlated with macrophage polarization. Upon overexpression of RBM15 in macrophages, there was an observed increase in the expression of M1-type macrophage markers CXCL9 and CXCL10, alongside a decrease in the expression of M2-type macrophage markers CCL13 and MRC1. Furthermore, there was an elevation in the expression of glycolytic enzymes GLUT1 and Hexokinase, as well as HIF1α, GAPDH, and PFKFB3 after RBM15 overexpression. Moreover, in vivo knockdown of RBM15 led to an amelioration of aortic aneurysm in the rat AD model. This knockdown also resulted in a reduction of the M1-type macrophage marker iNOS, while significantly increasing the expression of the M2-type macrophage marker CD206. In conclusion, our findings demonstrate that RBM15 upregulates glycolysis in macrophages, thus contributing to the progression of AD through the promotion of M1-type macrophage polarization. Conversely, downregulation of RBM15 suppresses M1-type macrophage polarization, thereby decelerating the advancement of AD. These results unveil potential novel targets for the treatment of AD.
摘要：
主动脉瘤和夹层（AD）是一种严重的心血管急症，死亡率高得惊人。最近的研究聚焦了AD患者中与m6A修饰相关的基因的过表达，将它们与炎性M1型巨噬细胞的存在联系起来。此外，糖酵解被广泛认为是炎性M1型巨噬细胞的关键特征，但是，将糖酵解和巨噬细胞功能联系起来以促进AD疾病进展的生物标志物尚未报道。我们对主动脉免疫细胞浸润进行了分析,巨噬细胞，使用生物信息学技术在AD患者中与m6A相关的生物标志物。随后，我们采用了RT-PCR的组合，WB,和免疫荧光分析以阐明M1型和M2型巨噬细胞表达的变化，以及糖酵解的标志，在关键生物标志物的过表达之后。通过创建具有前述关键生物标志物的敲低的AD大鼠模型,在体内进一步验证了这些发现。研究结果表明，m6A修饰的相关基因RBM15在AD样品中表达升高，并与巨噬细胞极化相关。在巨噬细胞中过表达RBM15后，观察到M1型巨噬细胞标志物CXCL9和CXCL10的表达增加,同时M2型巨噬细胞标志物CCL13和MRC1的表达减少.此外，糖酵解酶GLUT1和己糖激酶的表达升高，以及HIF1α，GAPDH,RBM15过表达后的PFKFB3。此外，体内敲低RBM15导致大鼠AD模型中主动脉瘤的改善。这种敲除还导致M1型巨噬细胞标记iNOS的减少，同时显着增加M2型巨噬细胞标志物CD206的表达。总之,我们的发现表明RBM15上调巨噬细胞的糖酵解，从而通过促进M1型巨噬细胞极化促进AD的进展。相反，RBM15的下调抑制M1型巨噬细胞极化，从而减缓了AD的发展。这些结果揭示了治疗AD的潜在新靶标。
