Research on the differential expression profiles of LncRNA and the calcification mechanism in human aortic smooth muscle cells induced by DPP4
10.3760/cma.j.cn311282-20250407-00177
- VernacularTitle:DPP4诱导人主动脉血管平滑肌细胞LncRNA差异表达谱和钙化机制研究
- Author:
Tongjie XU
1
;
Weidan LUO
1
;
Hao CHEN
1
;
Junlong ZHU
1
;
Hao YU
1
;
Huqiang HE
1
;
Yong LIU
1
Author Information
1. 西南医科大学附属医院血管外科，泸州 646000
- Publication Type:Journal Article
- Keywords:
Diabetes;
Dipeptidyl peptidase-4;
Vascular calcification;
Long non-coding RNA;
Matrix metalloproteinase-1
- From:
Chinese Journal of Endocrinology and Metabolism
2025;41(10):844-854
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the differential expression profiles of long non-coding RNAs(LncRNAs) and messenger RNAs(mRNAs) regulated by soluble dipeptidyl peptidase-4(sDPP4) during vascular smooth muscle cell calcification, and to explore the potential underlying calcification mechanisms.Methods:DPP4 levels in blood vessels and peripheral blood of diabetic patients were measured using Western blotting(WB) and real-time quantitative PCR(RT-qPCR). A cellular calcification model was established by treating human aortic vascular smooth muscle cells(HASMCs) with sDPP4. The effects of sDPP4 on HASMCs were assessed by WB, RT-qPCR, alizarin red staining, and calcium content determination. High-throughput sequencing was performed to analyze the differential expression profiles of LncRNA and mRNA following sDPP4 treatment. Among them, LncRNA ENST00000540293, which exhibited the most pronounced downregulation and was located adjacent to the matrix metalloproteinase-1(MMP-1) gene, was selected for further investigation. The osteogenic transdifferentiation of HASMCs after silencing LncRNA ENST00000540293 was evaluated using WB, RT-qPCR, alizarin red staining, and immunofluorescence-based cytoskeletal staining.Results:DPP4 expression was significantly elevated in both blood vessels and peripheral blood of diabetic patients. sDPP4 stimulation upregulated the protein levels of osteopontin(OPN) and runt-related transcription factor 2(RUNX2) in HASMCs, enhanced alizarin red staining, and increased intracellular calcium deposition. RNA sequencing revealed significant downregulation of LncRNA ENST00000540293 following sDPP4 exposure, while GO and pathway analysis indicated a marked increase in extracellular matrix binding activity(GO: 0050840). Silencing LncRNA ENST00000540293 suppressed α-smooth muscle actin(α-SMA) expression, promoted OPN and RUNX2 expression, increased calcification as shown by positive alizarin red staining, and cytoskeletal staining demonstrated osteogenic transdifferentiation of HASMCs, accompanied by a significant rise in MMP-1 protein level.Conclusion:sDPP4 promotes osteogenic transdifferentiation of HASMCs, potentially by downregulating LncRNA ENST00000540293. MMP-1 may be a potential target regulated by LncRNA ENST00000540293.
