P Dong1, W-J Liu, Z-H Wang. 1. Department of Cardiovascular Medicine, Beijing Aviation General Hospital, Beijing, China. kate-wzh@163.com.
Abstract
OBJECTIVE: To discover the mechanisms of miR-154 affecting myocardial fibrosis. PATIENTS AND METHODS: Human cardiac fibroblasts (CFs) were cultured in medium containing 10% serum for 48 h. The expression of miRNA-154 in human CFs was detected by Real-time quantitative polymerase chain reaction (qRT-PCR). The miRNA-154 mimics and inhibitors were synthesized and transfected into fibroblasts, respectively. Cell proliferation rate was determined by cell counting kit-8 (CCK8). Collagen I and collagen III, myofibroblast marker (a-SMA) and β-catenin were detected by Western blotting. Transwell migration assay was used to detect the changes of invasiveness of CFs. After the overexpression vector or siRNA of glycogen synthase kinase-3β (GSK-3β) was transfected into fibroblasts, we performed Western blot to detect a-SMA and β-catenin expression. RESULTS: MiR-154 was overexpressed in cardiomyocytes, and when miR-154 was inhibited, the expression of collagen I, collagen III, a-SMA, β-catenin, and the invasiveness of CFs decreased. Therefore, we considered that miR-154 could promote myocardial fibrosis by inhibiting the expression of GSK-3β. CONCLUSIONS: MiR-154 can inhibit GSK-3β expression by activating Wnt/β-catenin signaling pathway, which promotes myocardial fibrosis.
OBJECTIVE: To discover the mechanisms of miR-154 affecting myocardial fibrosis. PATIENTS AND METHODS: Human cardiac fibroblasts (CFs) were cultured in medium containing 10% serum for 48 h. The expression of miRNA-154 in human CFs was detected by Real-time quantitative polymerase chain reaction (qRT-PCR). The miRNA-154 mimics and inhibitors were synthesized and transfected into fibroblasts, respectively. Cell proliferation rate was determined by cell counting kit-8 (CCK8). Collagen I and collagen III, myofibroblast marker (a-SMA) and β-catenin were detected by Western blotting. Transwell migration assay was used to detect the changes of invasiveness of CFs. After the overexpression vector or siRNA of glycogen synthase kinase-3β (GSK-3β) was transfected into fibroblasts, we performed Western blot to detect a-SMA and β-catenin expression. RESULTS:MiR-154 was overexpressed in cardiomyocytes, and when miR-154 was inhibited, the expression of collagen I, collagen III, a-SMA, β-catenin, and the invasiveness of CFs decreased. Therefore, we considered that miR-154 could promote myocardial fibrosis by inhibiting the expression of GSK-3β. CONCLUSIONS:MiR-154 can inhibit GSK-3β expression by activating Wnt/β-catenin signaling pathway, which promotes myocardial fibrosis.
Authors: Nadia Saadat; Muraly Puttabyatappa; Venkateswaran R Elangovan; John Dou; Joseph N Ciarelli; Robert C Thompson; Kelly M Bakulski; Vasantha Padmanabhan Journal: Endocrinology Date: 2022-01-01 Impact factor: 4.736
Authors: Stuti Prakash; Andrea Mattiotti; Marc Sylva; Barbara J M Mulder; Alex V Postma; Maurice J B van den Hoff Journal: Mol Genet Genomic Med Date: 2019-02-05 Impact factor: 2.183