Abstract：
OBJECTIVE: Calcific aortic valve disease (CAVD) predominantly affects the elderly and currently lacks effective medical treatments. Nesfatin-1, a peptide derived from the cleavage of Nucleobindin 2, has been implicated in various calcification processes, both physiological and pathological. This study explores the impact of Nesfatin-1 on the transformation of aortic valve interstitial cells (AVICs) in CAVD.
RESULTS: In vitro experiments showed that Nesfatin-1 treatment mitigated the osteogenic differentiation of AVICs. Corresponding in vivo studies demonstrated a deceleration in the progression of CAVD. RNA-sequencing of AVICs treated with and without Nesfatin-1 highlighted an enrichment of the Ferroptosis pathway among the top pathways identified by the Kyoto Encyclopedia of Genes and Genomes analysis. Further examination confirmed increased ferroptosis in both calcified valves and osteoblast-like AVICs, with a reduction in ferroptosis following Nesfatin-1 treatment. Within the Ferroptosis pathway, ZIP8 showed the most notable modulation by Nesfatin-1. Silencing ZIP8 in AVICs increased ferroptosis and osteogenic differentiation, decreased intracellular Mn2+ concentration, and reduced the expression and activity of superoxide dismutase (SOD2). Furthermore, the silencing of SOD2 exacerbated ferroptosis and osteogenic differentiation. Nesfatin-1 treatment was found to elevate the expression of glutathione peroxidase 4 (GPX4) and levels of glutathione (GSH), as confirmed by Western blotting and GSH concentration assays.
CONCLUSIONS: In summary, Nesfatin-1 effectively inhibits the osteogenic differentiation of AVICs by attenuating ferroptosis, primarily through the GSH/GPX4 and ZIP8/SOD2 pathways.
RESULTS: In vitro experiments showed that Nesfatin-1 treatment mitigated the osteogenic differentiation of AVICs. Corresponding in vivo studies demonstrated a deceleration in the progression of CAVD. RNA-sequencing of AVICs treated with and without Nesfatin-1 highlighted an enrichment of the Ferroptosis pathway among the top pathways identified by the Kyoto Encyclopedia of Genes and Genomes analysis. Further examination confirmed increased ferroptosis in both calcified valves and osteoblast-like AVICs, with a reduction in ferroptosis following Nesfatin-1 treatment. Within the Ferroptosis pathway, ZIP8 showed the most notable modulation by Nesfatin-1. Silencing ZIP8 in AVICs increased ferroptosis and osteogenic differentiation, decreased intracellular Mn2+ concentration, and reduced the expression and activity of superoxide dismutase (SOD2). Furthermore, the silencing of SOD2 exacerbated ferroptosis and osteogenic differentiation. Nesfatin-1 treatment was found to elevate the expression of glutathione peroxidase 4 (GPX4) and levels of glutathione (GSH), as confirmed by Western blotting and GSH concentration assays.
CONCLUSIONS: In summary, Nesfatin-1 effectively inhibits the osteogenic differentiation of AVICs by attenuating ferroptosis, primarily through the GSH/GPX4 and ZIP8/SOD2 pathways.
摘要：
目的：钙化性主动脉瓣疾病（CAVD）主要影响老年人，目前缺乏有效的药物治疗。Nesfatin-1是一种来自核蛋白2裂解的肽，与各种钙化过程有关。生理和病理。本研究探讨Nesfatin-1对CAVD中主动脉瓣间质细胞（AVIC）转化的影响。
结果：体外实验表明Nesfatin-1处理减轻了AVIC的成骨分化。相应的体内研究表明CAVD的进展减速。用Nesfatin-1处理和不用Nesfatin-1处理的AVIC的RNA测序突出了由《京都基因和基因组百科全书》分析确定的顶级途径中铁凋亡途径的富集。进一步检查证实钙化瓣膜和成骨细胞样AVICs的铁性凋亡增加，Nesfatin-1治疗后铁性凋亡减少。在铁凋亡途径中，ZIP8显示了Nesfatin-1最显著的调节。在AVIC中沉默ZIP8可增加铁凋亡和成骨分化，细胞内Mn2+浓度降低，并降低超氧化物歧化酶(SOD2)的表达和活性。此外，SOD2的沉默加剧了铁性凋亡和成骨分化。Nesfatin-1治疗可提高谷胱甘肽过氧化物酶4（GPX4）的表达和谷胱甘肽（GSH）的水平，通过Western印迹和GSH浓度测定证实。
结论：总之，Nesfatin-1通过减弱铁性凋亡有效抑制AVICs的成骨分化,主要通过GSH/GPX4和ZIP8/SOD2途径。
结果：体外实验表明Nesfatin-1处理减轻了AVIC的成骨分化。相应的体内研究表明CAVD的进展减速。用Nesfatin-1处理和不用Nesfatin-1处理的AVIC的RNA测序突出了由《京都基因和基因组百科全书》分析确定的顶级途径中铁凋亡途径的富集。进一步检查证实钙化瓣膜和成骨细胞样AVICs的铁性凋亡增加，Nesfatin-1治疗后铁性凋亡减少。在铁凋亡途径中，ZIP8显示了Nesfatin-1最显著的调节。在AVIC中沉默ZIP8可增加铁凋亡和成骨分化，细胞内Mn2+浓度降低，并降低超氧化物歧化酶(SOD2)的表达和活性。此外，SOD2的沉默加剧了铁性凋亡和成骨分化。Nesfatin-1治疗可提高谷胱甘肽过氧化物酶4（GPX4）的表达和谷胱甘肽（GSH）的水平，通过Western印迹和GSH浓度测定证实。
结论：总之，Nesfatin-1通过减弱铁性凋亡有效抑制AVICs的成骨分化,主要通过GSH/GPX4和ZIP8/SOD2途径。
