PDF(Pubmed)
Abstract：
UNASSIGNED: Cellular senescence, macrophages infiltration, and vascular smooth muscle cells (VSMCs) osteogenic transdifferentiation participate in the pathophysiology of vascular calcification in chronic kidney disease (CKD). Senescent macrophages are involved in the regulation of inflammation in pathological diseases. In addition, senescent cells spread senescence to neighboring cells via Interferon-induced transmembrane protein3 (IFITM3). However, the role of senescent macrophages and IFITM3 in VSMCs calcification remains unexplored.
UNASSIGNED: To explore the hypothesis that senescent macrophages contribute to the calcification and senescence of VSMCs via IFITM3.
UNASSIGNED: Here, the macrophage senescence model was established using Lipopolysaccharides (LPS). The VSMCs were subjected to supernatants from macrophages (MCFS) or LPS-induced macrophages (LPS-MCFS) in the presence or absence of calcifying media (CM). Senescence-associated β-galactosidase (SA-β-gal), Alizarin red (AR), immunofluorescent staining, and western blot were used to identify cell senescence and calcification.
UNASSIGNED: The expression of IFITM3 was significantly increased in LPS-induced macrophages and the supernatants. The VSMCs transdifferentiated into osteogenic phenotype, expressing higher osteogenic differentiation markers (RUNX2) and lower VSMCs constructive makers (SM22α) when cultured with senescent macrophages supernatants. Also, senescence markers (p16 and p21) in VSMCs were significantly increased by senescent macrophages supernatants treated. However, IFITM3 knockdown inhibited this process.
UNASSIGNED: Our study showed that LPS-induced senescence of macrophages accelerated the calcification of VSMCs via IFITM3. These data provide a new perspective linking VC and aging, which may provide clues for diagnosing and treating accelerated vascular aging in patients with CKD.
UNASSIGNED: To explore the hypothesis that senescent macrophages contribute to the calcification and senescence of VSMCs via IFITM3.
UNASSIGNED: Here, the macrophage senescence model was established using Lipopolysaccharides (LPS). The VSMCs were subjected to supernatants from macrophages (MCFS) or LPS-induced macrophages (LPS-MCFS) in the presence or absence of calcifying media (CM). Senescence-associated β-galactosidase (SA-β-gal), Alizarin red (AR), immunofluorescent staining, and western blot were used to identify cell senescence and calcification.
UNASSIGNED: The expression of IFITM3 was significantly increased in LPS-induced macrophages and the supernatants. The VSMCs transdifferentiated into osteogenic phenotype, expressing higher osteogenic differentiation markers (RUNX2) and lower VSMCs constructive makers (SM22α) when cultured with senescent macrophages supernatants. Also, senescence markers (p16 and p21) in VSMCs were significantly increased by senescent macrophages supernatants treated. However, IFITM3 knockdown inhibited this process.
UNASSIGNED: Our study showed that LPS-induced senescence of macrophages accelerated the calcification of VSMCs via IFITM3. These data provide a new perspective linking VC and aging, which may provide clues for diagnosing and treating accelerated vascular aging in patients with CKD.
摘要：
■细胞衰老，巨噬细胞浸润，血管平滑肌细胞(VSMCs)成骨转分化参与慢性肾脏病(CKD)血管钙化的病理生理过程。衰老巨噬细胞参与病理疾病中炎症的调节。此外,衰老细胞通过干扰素诱导的跨膜蛋白3（IFITM3）将衰老传播到相邻细胞。然而,衰老巨噬细胞和IFITM3在VSMC钙化中的作用仍有待探索。
■探索衰老巨噬细胞通过IFITM3促进VSMC钙化和衰老的假设。
■这里，采用脂多糖(LPS)建立巨噬细胞衰老模型。在存在或不存在钙化培养基(CM)的情况下,使VSMC经受来自巨噬细胞(MCFS)或LPS诱导的巨噬细胞(LPS-MCFS)的上清液。衰老相关β-半乳糖苷酶（SA-β-gal），茜素红（AR），免疫荧光染色，用免疫印迹法鉴定细胞衰老和钙化。
■在LPS诱导的巨噬细胞和上清液中，IFITM3的表达显着增加。VSMC转分化为成骨表型，当与衰老的巨噬细胞上清液一起培养时，表达较高的成骨分化标记（RUNX2）和较低的VSMC建设性标记（SM22α）。此外，经衰老巨噬细胞上清液处理，VSMC中的衰老标记（p16和p21）显着增加。然而,IFITM3敲低抑制了该过程。
■我们的研究表明，LPS诱导的巨噬细胞衰老通过IFITM3加速了VSMC的钙化。这些数据提供了连接VC和老龄化的新视角，为CKD患者加速血管老化的诊断和治疗提供线索。
■探索衰老巨噬细胞通过IFITM3促进VSMC钙化和衰老的假设。
■这里，采用脂多糖(LPS)建立巨噬细胞衰老模型。在存在或不存在钙化培养基(CM)的情况下,使VSMC经受来自巨噬细胞(MCFS)或LPS诱导的巨噬细胞(LPS-MCFS)的上清液。衰老相关β-半乳糖苷酶（SA-β-gal），茜素红（AR），免疫荧光染色，用免疫印迹法鉴定细胞衰老和钙化。
■在LPS诱导的巨噬细胞和上清液中，IFITM3的表达显着增加。VSMC转分化为成骨表型，当与衰老的巨噬细胞上清液一起培养时，表达较高的成骨分化标记（RUNX2）和较低的VSMC建设性标记（SM22α）。此外，经衰老巨噬细胞上清液处理，VSMC中的衰老标记（p16和p21）显着增加。然而,IFITM3敲低抑制了该过程。
■我们的研究表明，LPS诱导的巨噬细胞衰老通过IFITM3加速了VSMC的钙化。这些数据提供了连接VC和老龄化的新视角，为CKD患者加速血管老化的诊断和治疗提供线索。
