Pang Honggang, Fu Bowen, Wang Panxing, Meng Yan, Xie Peng, Hu Xilong, Ma Qiang
Department of Peripheral Vascular Diseases, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China.
Hepatobiliary and Vascular Surgery, 521 Hospital of Norinco Group, Xi'an, China.
J Bioenerg Biomembr. 2025 Jul 11. doi: 10.1007/s10863-025-10065-y.
Abdominal aortic aneurysm (AAA) is a life-threatening vascular disease characterized by aortic wall degeneration and inflammation. The molecular mechanisms underlying AAA development remain unclear. Wilms tumor 1-associated protein (WTAP) has been implicated in various biological processes, but its role in AAA pathogenesis, particularly in cardiomyocyte regulation, has not been fully explored. Quantitative real-time PCR (qRT-PCR) was performed to detect the mRNA levels of WTAP and proprotein convertase subtilisin/kexin type 9 (PCSK9). Western blotting assay was used to analyze protein expression. Cell viability, proliferation, senescence, apoptosis, ferroptosis, and inflammation were assessed using cell counting kit-8 assay, 5-Ethynyl-2'-deoxyuridine assay, SA-β-gal staining, flow cytometry, fluorometric assay, colorimetric method, and enzyme-linked immunosorbent assay. The association among PCSK9, WTAP, and IGF2BP2 was analyzed using RNA immunoprecipitation assay and dual-luciferase reporter assay. WTAP expression was upregulated in AAA and angiotensin II (Ang II)-induced human aortic smooth muscle cells (HASMCs). Ang II treatment inhibited HASMC proliferation and induced senescence, apoptosis, ferroptosis, and NLRP3 inflammasome-mediated inflammation. However, these effects were mitigated by WTAP knockdown. In addition, PCSK9 expression was increased in AAA, and WTAP stabilized PCSK9 mRNA expression in an IGF2BP2-dependent manner. Moreover, WTAP overexpression promoted senescence, apoptosis, ferroptosis, and inflammation by regulating PCSK9 in Ang II-induced HASMCs. WTAP silencing protected HASMCs from Ang II-induced senescence, apoptosis, ferroptosis, and inflammation by regulating PCSK9, suggesting a potential therapeutic target for AAA treatment.
腹主动脉瘤(AAA)是一种危及生命的血管疾病,其特征为主动脉壁退变和炎症。AAA发生发展的分子机制仍不清楚。肾母细胞瘤1相关蛋白(WTAP)参与了多种生物学过程,但其在AAA发病机制中的作用,尤其是在心肌细胞调节方面,尚未得到充分研究。采用定量实时PCR(qRT-PCR)检测WTAP和前蛋白转化酶枯草溶菌素/kexin 9型(PCSK9)的mRNA水平。利用蛋白质免疫印迹法分析蛋白表达。使用细胞计数试剂盒-8法、5-乙炔基-2'-脱氧尿苷法、SA-β-半乳糖苷酶染色、流式细胞术、荧光法、比色法和酶联免疫吸附测定法评估细胞活力、增殖、衰老、凋亡、铁死亡和炎症。采用RNA免疫沉淀测定法和双荧光素酶报告基因测定法分析PCSK9、WTAP和IGF2BP2之间的关联。WTAP在AAA和血管紧张素II(Ang II)诱导的人主动脉平滑肌细胞(HASMCs)中表达上调。Ang II处理抑制HASMC增殖并诱导衰老、凋亡、铁死亡和NLRP3炎性小体介导的炎症。然而,WTAP敲低可减轻这些作用。此外,PCSK9在AAA中表达增加,WTAP以IGF2BP2依赖的方式稳定PCSK9 mRNA表达。此外,WTAP过表达通过调节Ang II诱导的HASMCs中的PCSK9促进衰老、凋亡、铁死亡和炎症。WTAP沉默通过调节PCSK9保护HASMCs免受Ang II诱导的衰老、凋亡、铁死亡和炎症,提示其可能是AAA治疗的潜在靶点。