Highlights
- •Calcific aortic valve disease is an important and common cardiovascular disease.
- •long noncoding TSI expression was upregulated in calcified aortic valves.
- •lncTSI alleviated aortic valve calcification through negative regulation of the TGF-β/Smad3 pathway.
- •These data suggest that long noncoding TSI is a potential therapeutic target.
Abstract
Introduction
Calcific aortic valve disease (CAVD) is an active and cellular-driven fibrocalcific
process characterised by differentiation of valve interstitial cells (VICs) towards
an osteogenic-like phenotype. A recently identified lncRNA, lncTSI, has been reported to inhibit fibrogenesis through transforming growth factor (TGF)-β/Smad3
pathway. Here, the present study aimed to investigate the role of lncTSI in CAVD.
Methods
The effect of TGF-β1 on lncTSI of VICs was measured. TGF-β1, RUNX2 and collagen I expression between calcified aortic
valve tissue and normal samples by immunohistochemistry and western blotting. Human
VICs were cultured and treated with TGF-β1. SiRNA and pcDNA3.1-lncTSI plasmid transfection
were used to silence and overexpress lncTSI in VICs for 48 h, Smads phosphorylation, RUNX2 and collagen I expression were then
verified by western blotting. In ApoE−/− mice fed with 0.25 % high-cholesterol diet, AAV2-lncTSI were injected intravenously to observe their effect on the formation of aortic valve
calcification.
Results
lncTSI was highly expressed in VICs treated with TGF-β1. lncTSI was transcriptionally regulated by Smad3 and reversely inhibited TGF-β1-induced Smad3
phosphorylation and downregulated profibrotic gene expression. Silencing lncTSI increased TGF-β1–induced Smad3 phosphorylation, and subsequently, upregulated RUNX2
and collagen I expressions in VICs. While overexpression of lncTSI reversed the production of RUNX2 and collagen I in VICs. In a mouse CAVD model of
24 week 0.25 % high-cholesterol diet feeding, overexpression of lncTSI significantly reduced calcium deposition, RUNX2, pSmad3, and collagen I expression
in aortic valve leaflets, with less aortic valve stenosis.
Conclusions
The novel findings of present study suggested that lncTSI alleviated aortic valve calcification through negative regulation of the TGF-β/Smad3
pathway. The results may help elucidate new diagnostic and therapeutic targets to
prevent CAVD progression.
Abbreviations:
CAVD (calcific aortic valve disease), VICs (valve interstitial cells), TGF-β (transforming growth factor β), lncTSI (long noncoding TSI), IL (interleukin), COL1A1 (type I collagen), RUNX2 (Runt-related Transcription Factor 2), FBS (fetal bovine serum), GAPDH (glyceraldehyde-3-phosphate dehydrogenase), H&E staining (haematoxylin-eosin staining), PBS (phosphate buffer solution), ApoE−/− (apolipoprotein E–deficient), ROS (reactive oxygen species)Keywords
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Article info
Publication history
Published online: October 20, 2022
Accepted:
October 17,
2022
Received:
August 18,
2022
Identification
Copyright
© 2022 Published by Elsevier Inc.
