Exosomal STAT1 derived from high phosphorus‑stimulated vascular endothelial cells induces vascular smooth muscle cell calcification via the Wnt/β‑catenin signaling pathway

Abstract

Vascular calcification is commonly observed in chronic kidney disease. The mechanism of how the calcification signal from endothelial cells is transmitted to vascular smooth muscle cells (VSMCs) remains unknown. The aim of the present study was to investigate whether exosomes from HUVECs (HUVEC-Exos) could regulate VSMC calcification and its potential signaling pathway. HUVEC-Exos were isolated from HUVECs under no phosphorus (NP) and high phosphorus (HP) conditions. Alizarin Red S staining and calcium (Ca) content analysis were carried out to detect calcification in VSMCs. Proteomics analysis was carried out to detect the differential expression of exosomal proteins. Protein and mRNA levels were measured by western blot analysis and reverse transcription-quantitative PCR (RT-qPCR). Exosomes derived from HP-HUVECs promoted the calcification of VSMCs, as assessed by Alizarin Red S staining, alkaline phosphatase activity assays, Ca content measurements and the increased expression of runt-related transcription factor 2 and osteopontin. Proteomic analysis detected the upregulation of STAT1 in HP-exosomes from HUVECs (HUVEC-Exos) compared with NP-HUVEC-Exos, which was also confirmed by western blot analysis and RT-qPCR. Inhibition of STAT1 expression in VSMCs using fludarabine or knockdown of STAT1 expression using small interfering RNA alleviated the calcification of VSMCs. Furthermore, lithium chloride (Wnt activator) reversed the protective effect of STAT1 inhibition on VSMC calcification, while Dickkopf-1 (Wnt inhibitor) exerted the opposite effect, suggesting that activation of the Wnt/β-catenin signaling pathway was involved in STAT1-mediated VSMC calcification. In conclusion, the present results indicated that exosomal STAT1 derived from HP-treated HUVECs could promote VSMC calcification, and activation of the Wnt/β-catenin pathway may be a potential mechanism of the VSMC calcification promoted by exosomes.

Inhibition of complement C3 prevents osteoarthritis progression in guinea pigs by blocking STAT1 activation

“…To verify our findings, we used fludarabine, which has shown to be a specific STAT1 inhibitor in previous studies 30 , 31 . It is currently used clinically as chemotherapy to treat chronic lymphocytic leukemia 32 .…”

Adipose-Derived Stem Cell Exosomes Antagonize the Inhibitory Effect of Dihydrotestosterone on Hair Follicle Growth by Activating Wnt/β-Catenin Pathway

“…Deleted Deleted However, which component in ADSC-Exos may play a role in activating Wnt/βcatenin signaling pathway still remains unspecified and needs further exploration. Even though numerous previous studies have shown that a variety of effective substances (e.g., miR-424-5p, miR-1260b, miR-127-3p, miR-218-5p, miR-22-5p, miR-181a-5p, and miR-148b-3p [54,58]; protein Hic-5, UBR2, and STAT1 [59][60][61][62][63][64][65]) in exosomes from the different sources can regulate the Wnt/β-catenin signaling pathway, single miRNA or protein from exosomes might not completely recapitulate the regulatory effects. Therefore, in the present study, instead of exosomal miRNAs or proteins, we used exosomes as a whole to investigate the therapeutic effects on AGA.…”

Pathogenesis and Mechanism of Uremic Vascular Calcification

“…Qin et al demonstrated that STAT1-rich exosomes secreted by HP-treated HUVECs are delivered to VSMCs, thus promoting VSMC calcification through the activation of Wnt/β-catenin signaling. This result indicates that HP-HUVEC-Exo has calcium-promoting effects [ 87 ]. In addition to target proteins, miRNAs encapsulated in EC-EVs have been demonstrated to modulate blood vessels in CKD-MBD, with miR-143-3p and miR-145-5p inhibiting VC and miR-221-3p and miR-222-3p enhancing VC [ 88 ].…”