Effects of sFRP3 overexpression on the activation and proliferation of murine cardiac fibroblasts
Objective To explore the role of secreted frizzled-related protein 3 (sFRP3), a regulator of the Wnt signaling pathway, in the activation and proliferation of murine cardiac fibroblasts (CFs).Methods Neonatal mice aged 1-3 days were obtained for surgical procedures to collect heart tissues.After digestion, CFs were isola-ted and cultured.Transforming growth factor-beta 1 (TGF-β1) stimulation was used to induce activation and prolif-eration in CFs after they adhered to the culture dish.Once the model was confirmed, experimental and control groups were transfected with sFRP3 overexpression plasmids and empty plasmids for 24-48 hours.Expression lev-els of sFRP3, Periostin (POSTN), Type Ⅰ collagen (Collagen Ⅰ), and proliferating cell nuclear antigen (PC-NA) were assessed at the molecular level using Western blot and qRT-PCR.Changes in cell proliferation capacity were examined using MTT, CCK-8, and EdU staining methods.Results In the TGF-β1-induced activation and proliferation model of CFs, compared to the control group, the model group exhibited decreased expression of sFRP3 protein and mRNA, while the expression of activation and proliferation-related proteins PCNA, POSTN, and Collagen Ⅰ was upregulated.Furthermore, in CFs overexpressing sFRP3 through plasmid transfection, the protein and mRNA expression of PCNA, POSTN, and Collagen Ⅰ decreased compared to the empty vector group.MTT, CCK-8 , and EdU experiments indicated a significant decrease in the proliferative activity of CFs in the sFRP3 over-expression group compared to the empty vector group.Conclusion Overexpression of sFRP3 markedly inhibits the activation and proliferation of CFs, suggesting that sFRP3 may be a key gene involved in the regulation of CF acti-vation and proliferation.
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