Reduced exosome miR‐425 and miR‐744 in the plasma represents the progression of fibrosis and heart failure

Figures

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  Figure 1. Exosomal miRNAs level was altered in the patients with HF. Exosomes were extracted from the peripheral blood samples of 31 HF patients and paired healthy controls. The level of 9 candidate miRNAs was detected qRT‐PCR. Results were analyzed by student's t‐test and p < 0.05 was considered statistically significant. *p < 0.05, ***p < 0.001.

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  Figure 2. In vitro silencing of miR‐425 and miR‐744 caused fibrogenesis in cultured cardiac fibroblasts. NHCF‐A cells were treated with Ang II and the expression of collagen 1 and α‐SMA was determined by immunoblotting (A). The level of miR‐21, miR‐425 and miR‐744 was detected by qRT‐PCR(B). MiR‐425 and miR‐744 inhibitors were transient transfected into NFCF‐A cells for 48 h and total RNA was extracted for miRNA(C) and mRNA (D, E) detection. The protein level of collagen 1 and α‐SMA was determined by immunoblotting.

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  Figure 3. Forced expression of miR‐425 and miR‐744 attenuated Ang II induced fibrogenesis in cultured cardiac fibroblasts. NHCF‐A cells were transfected with miR‐425 or miR‐744 mimic, the transfection efficiency of miR‐425 and miR‐744 was confirmed by qRT‐PCR(A). The mRNA level of Col1α1 and Col3α1 was determined by RT‐PCR(B) and protein level of collagen 1 and α‐SMA was determined by immunoblotting.

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  Figure 4. TGFB1 is a direct target of miR‐425 and miR‐744. (A) Predicted target site of miR‐425 and miR‐744 in the 3′UTR of TGFB1 mRNA. (B) HEK293T cells were transfected with TGFB1 luciferase reporter vector with miRNA mimic or inhibitor. 48 h after transfection, cells were lysed and luciferase activity was detected. Results were analyzed by student's t‐test and p < 0.05 was considered statistically significant. *p < 0.05, **P < 0.01. (C) NHCF‐A cells were transfected with miRNA mimic or inhibitor for 48 h, the expression of TGFβ1 was determined by immunoblotting. The band densities were quantified by Quantity One (BioRad), and results were analyzed by t‐test. *p < 0.05, **P < 0.01.