Levocarnitine regulates the growth of angiotensin II-induced myocardial fibrosis cells via TIMP-1

Figures

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  Figure 1. Expression of MF-related markers in AngII-induced cells using qRT-PCR and western blotting. (a) mRNA expression of Axl, α-SMA, MMP3, STAT4, and TIMP-1 after AngII treatment for 24 h determined using qRT-PCR. (b) Protein expression of Axl, α-SMA, MMP3, STAT4, and TIMP-1 after AngII treatment for 24 h determined using western blotting. *P < 0.05 vs control. MF, myocardial fibrosis.

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  Figure 2. Expression of fibroblast-related markers in AngII-induced cells and cell transfection efficiency. (a) mRNA expression of FSP1, COL1A1, COL3A1, and TGF-β1 after AngII treatment for 24 h determined using qRT-PCR. *P < 0.05 vs control. (b) mRNA expression of TIMP-1 after transfection with si-TIMP-1. (c) Protein expression of TIMP-1 after transfection with si-TIMP-1. (d) mRNA expression of TIMP-1 after transfection with TIMP-1-OE plasmid. (e) Protein expression of TIMP-1 after transfection with TIMP-1-OE plasmid. *P < 0.05 vs blank, # P < 0.05 vs si-TIMP-1(1). TIMP‐1, tissue inhibitor of metalloproteinases‐1; MF, myocardial fibrosis; si, small interfering RNA; OE, over-expression.

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  Figure 3. Cell viability of H9C2 cells after si-TIMP-1 and TIMP-1-OE transfection for 24 h (a), 48 h (b), 72 h (c), and 96 h (d). *P < 0.05 vs control, # P < 0.05 vs MF, $ P < 0.05 vs MF + LC. TIMP‐1, tissue inhibitor of metalloproteinases‐1; MF, myocardial fibrosis; siRNA, small interfering RNA; OE, over-expression; LC, levocarnitine.

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  Figure 4. Effects of TIMP-1 on cell apoptosis and migration of H9C2 cells induced by AngII. (a) Cell apoptosis in the different groups determined using flow cytometry. (b) Cell migration in the different groups assessed using tanswell assay. *P < 0.05 vs control, # P < 0.05 vs MF, $ P < 0.05 vs MF + LC. TIMP‐1, tissue inhibitor of metalloproteinases‐1; MF, myocardial fibrosis; siRNA, small interfering RNA; OE, over-expression; LC, levocarnitine.

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  Figure 5. Effects of TIMP-1 on the expression of oxidative stress-related genes, MF-related genes, and apoptosis-related genes determined using qRT-PCR. Relative mRNA expression of Axl (a), AT1R (b), α-SMA (c), Collagen III (d), Caspase 3 (e), p53 (f), Bcl2 (g), MMP3 (h), and STAT4 (i). *P < 0.05 vs control, # P < 0.05 vs MF, $ P < 0.05 vs MF + LC. TIMP‐1, tissue inhibitor of metalloproteinases‐1; MMP, matrix metalloproteinase; Axl, AXL receptor tyrosine kinase; AT1R, angiotensin II receptor type 1; α-SMA, α-smooth muscle actin; MF, myocardial fibrosis; siRNA, small interfering RNA; OE, over-expression, LC, levocarnitine.

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  Figure 6. Effects of TIMP-1 on protein expression levels in H9C2 cells determined using western blotting. (a) Representative images of western blotting. The protein expression of TIMP-1 (b), Caspase‑3 (c), and p53 (d). *P < 0.05 vs control, # P < 0.05 vs MF, $ P < 0.05 vs MF + LC. TIMP‐1, tissue inhibitor of metalloproteinases‐1; MF, myocardial fibrosis; siRNA, small interfering RNA; OE, over-expression, LC, levocarnitine.

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  Figure A1. Effects of empty plasmids and transfection reagents on the cell viability of H9C2 cells after culturing for 24, 48, 72, and 96 h using CCK-8.