Mechanism Study of CTHRC1 Promoting Pathological Myocardial Remodeling via Oxidative Stress Pathway
LI Nan
FU Heqing
CHU Shiqi
ZHANG Shu
NIE Huijuan
DONG Yu
LIU Tianlong
Abstract：Objective:To observe the effect of overexpression of collagen triple helix repeat containing-1(CTHRC1)in vitro on pathological myocardial remodeling induced by angiotensin Ⅱ(Ang Ⅱ)and the related mechanisms.Methods:The rat H9c2 cardiomyocytes cultured in vitro were divided into the GFP group(transfected with negative control GFP-AAV9 type gene adenovirus 4.8×1010 PFU/mL)and the CTHRC1 group(transfected with CTHRC1-AAV9 type gene adenovirus 2.4×1010 PFU/mL).The GFP group was further divided into the Vehicle-GFP group(transfected with negative control GFP-AAV9 type gene adenovirus 4.8×1010 PFU/mL)and the GFP-AngⅡ group(transfected with negative control GFP-AAV9 type gene adenovirus 4.8×1010 PFU/mL,AngⅡ 10 μmol/L).the CTHRC1 group was divided into the Vehicle-CTHRC1 group(transfected with CTHRC1-AAV9 type gene adenovirus 2.4×1010 PFU/mL)and the CTHRC1-AngⅡ group(transfected with CTHRC1-AAV9 type gene adenovirus 2.4×1010 PFU/mL,AngⅡ 10 μmol/L).The expression of CTHRC1 protein was detected by Western Blot.The surface area of H9c2 cells was evaluated by phalloidin staining.After 10％sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE),the gel was stained with Coomassie Brilliant blue,cut,and protein identification was performed using high-resolution mass spectrometry.Gene Ontology(GO)gene,and set enrichment analysis(GSEA)functional enrichment analysis was conducted for the differentially expressed genes.The activities of superoxide dismutase(SOD),catalase(CAT),and malondialdehyde(MDA)were detected using kits.Results:Compared with the Vehicle-GFP group,the surface area of H9c2 cells in the GFP-AngⅡ group increased(P<0.001).Compared with the Vehicle-CTHRC1 group,the surface area of H9c2 cells in the CTHRC1-AngⅡgroup increased(P<0.001).Compared with the GFP-AngⅡ group,the surface area of H9c2 cells in the CTHRC1-AngⅡ group increased(P<0.001).Compared with the Vehicle group,the expression of H9c2 cardiomyocytes in the AngⅡ group decreased around 75 kDa.Compared with the GFP group,the CTHRC1 group showed 31 proteins significantly downregulated and 30 proteins significantly upregulated(P<0.05).For the significantly downregulated proteins,the GO analysis showed that signaling pathways such as glutathione metabolism and protein stability were significantly enriched.For the significantly downregulated proteins,the GSEA analysis showed that the glutathione metabolism pathway was significantly enriched.Compared with the GFP-AngⅡ group,the H9c2 cardiomyocytes in the Vehicle-GFP group showed lower SOD and CAT activities and higher MDA activity(P<0.05 or P<0.001).Compared with the Vehicle-CTHRC1 group,the H9c2 cardiomyocytes in the CTHRC1-AngⅡ group showed lower SOD and CAT activities and higher MDA activity(P<0.01 or P<0.001).Compared with the GFP-AngⅡ group,the H9c2 cardiomyocytes in the CTHRC1-AngⅡ group showed lower SOD and CAT activities and higher MDA activity(P<0.05 or P<0.001).Conclusion:Overexpression of CTHRC1 in vitro could aggravate pathological myocardial remodeling induced by AngⅡ,and its function was related to the inhibition of the glutathione metabolism pathway.
Keywords：myocardial remodelingcollagen triple helix repeat containing-1oxidative stressenrichment analysisglutathione metabolismexperimental study
Publication Date：2026-03-25
Online Publishing Date：2026-04-07(First online date of this platform, not the publication date of the document)
Pages：9( 869-877 )