Inhibitory Effects of the Slit Guidance Ligand 1-3’ Untranslated Region on the Fibrotic Phenotype of Cardiac Fibroblasts
10.13471/j.cnki.j.sun.yat-sen.univ（med.sci）.2025.0311
- VernacularTitle:狭缝引导配体1-3’非翻译区抑制心肌成纤维细胞纤维化表型的作用
- Author:
Ya WANG
1
;
Huayan WU
1
;
Yuan GAO
2
;
Rushi WU
1
;
Peiying GUAN
1
;
Hui LI
3
;
Juntao FANG
4
;
Zhixin SHAN
1
Author Information
1. School of Medicine， South China University of Technology， Guangzhou 510006， China
2. Guangdong Provincial People’s Hospital//Guangdong Academy of Medical Sciences//Southern Medical University， Guangzhou 510080， China
3. Department of Clinical Laboratory， Guangdong Provincial People’s Hospital//Guangdong Academy of Medical Sciences//Southern Medical University， Guangzhou 510080， China
4. Department of Cardiology， Guangdong Cardiovascular Institute//Guangdong Provincial People’s Hospital//Guangdong Academy of Medical Sciences//Southern Medical University， Guangzhou 510080， China
- Publication Type:Journal Article
- Keywords:
myocardial fibrosis;
slit guidance ligand 1（Slit1）;
3’untranslated region （3’UTR）;
microRNA;
cardiac fibroblasts
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2025;46(3):466-474
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo study the regulatory effect of the partial sequence within the 3’ untranslated region （3’UTR） of slit guidance ligand 1 （Slit1） （Slit1-3’UTR） on the fibrotic phenotypes of cardiac fibroblasts （CFs） and its potential mechanism. MethodsThe adenovirus vector was used to overexpress the 1526nt sequence of Slit1-3’UTR in ICR neonatal mouse CFs （mCFs）. The expression of fibrosis-related genes in mCFs， such as collagen type 1 alpha1（COL1A1）， collagen type 3 alpha3 （COL3A1） and alpha smooth muscle actin （α-SMA） were detected by Western blot assay. The effect of Slit1-3’UTR 1526nt on the proliferation and migration of mCFs was assessed by EdU staining and Trans-well assays. Angiotensin Ⅱ （Ang Ⅱ） was used to treat mCFs， and the impact of Slit1-3’UTR 1526nt on the fibrotic phenotypes of Ang Ⅱ-induced mCFs was evaluated. After overexpression of Slit1-3’UTR 1526nt， miR-34a-5p mimic was transfected into mCFs， followed by actinomycin D treatment to detect the mRNA stability of Slit1-3’UTR 1526nt， and the levels of miR-34a-5p and its target gene SIRT1（si-SIRT1） in mCFs were determined. The effects of miR-34a-5p and small interfering RNA targeting SIRT1 on the Slit1-3’UTR 1526nt-mediated regulation of fibrotic phenotypes were also determined. ResultsAdenovirus-mediated overexpression of Slit 1-3’UTR 1526nt was achieved in mCFs. Overexpression of Slit 1-3’UTR 1526nt markedly inhibited the expression of the fibrosis-related genes， proliferation and migration of mCFs and fibrotic phenotypes of Ang Ⅱ. The results of actinomycin D assay showed that miR-34a-5p inhibited the stability of Slit1-3’UTR 1526nt in mCFs， while the level of miR-34a-5p was reduced in mCFs with overexpression of Slit1-3’UTR 1526nt. Transfection of miR-34a-5p promoted the fibrotic phenotypes， and reversed the inhibitory effect of Slit1-3’UTR 1526nt on the fibrotic phenotypes of mCFs. Overexpression of Slit1-3’UTR 1526nt significantly increased the level of miR-34a-5p target gene SIRT1 in mCFs. Transfection of miR-34a-5p and si-SIRT1 consistently reversed the inhibitory effects of Slit1-3’UTR 1526nt on the fibrotic phenotypes of mCFs. ConclusionSlit1-3’UTR1526nt inhibits the fibrotic phenotypes of mCFs by binding to miR-34a-5p and increasing the expression of its target gene of SIRT1.
