Pared with control group (Fig. 2f ).Relugolix solubility miR-21 overexpression in cardiac fibrotic
Pared with control group (Fig. 2f ).miR-21 overexpression in cardiac fibrotic remodeling and fibroblastTo gain the expression of miR-21 in cardiac fibrotic remodeling and fibroblast. One-step qRT-PCR assay demonstrated that miR-21 expression in the rat model group and AF group were increased significantly compared with the control group (Fig. 3a). Moreover, One-step qRT-PCR found that miR-21 expression increasedABFig. 3 miR-21 increased in PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/27196668 activated cardiac fibroblasts and fibrotic remodeling. a The miR-21 expression was assayed in rat and human cardiac fibrosis tissue. One Step Real-time PCR showed that miR-21 was significantly up-regulated in cardiac fibrosis tissue compared to vehicle tissue. Moreover, One Step Real-time PCR showed that miR-21 was significantly up-regulated in AF tissue compared to SR tissue. b The miR-21 expression was assayed in CFs stimulated with TGF-1. One Step Real-time PCR showed that miR-21 was significantly increased in activated CFs compared to control group cell. Data are representative of at least three separate experiments. *P < 0.05 versus control or Vehicle or SRCao et al. BMC Cardiovascular Disorders (2017) 17:Page 7 ofduring activated cardiac fibroblasts, which was treated with TGF-1 compared with control group (Fig. 3b).Knockdown of miR-21 inhibits the cardiac fibroblast proliferationTo explore the roles of miR-21 in regulating cardiac fibroblasts cell proliferation, we tested the effect of miR21 down-expression on the cell proliferation. By Onestep qRT-PCR, we found that the expression of miR-21 significantly decreased in the cells transfected with miR21 inhibitor compared with NC and vehicle (Fig. 4a). The cardiac fibroblast that was transfected with miR-inhibitor had a significantly lower proliferation than the NC and vehicle (Fig. 4b). Cell cycle distribution was analyzed by Flow cytometry analysis showed that antiproliferative activity of miR-21 inhibitor was possibly due to induce G2/M phase arrest (Fig. 4c). These results suggested that down-expression of miR-21 inhibits the cardiac fibroblasts proliferation.miR-21 negative control CADM1 expressionTo further determine the underlying mechanism of miR21 during cardiac fibroblast activation. We began our studies interested in probing possible regulation of theABCFig. 4 Knockdown of miR-21 inhibits the cardiac fibroblasts proliferation. a One Step Real-time PCR showed that transfection miR-21 inhibitor decreased the expression of miR-21 in cardiac fibroblasts. miR-21 inhibitor group is decreased significantly compare with vehicle group. b Cell viability was determined using MTT assay. 3 ?105 cardiac fibroblasts cells were seeded in 96-well plates and transfected with 60 nM miR-21 inhibitor (24 h, 48 h). Knockdown of miR-21 caused a significant inhibition of cell activation and proliferation in TGF-1-treated cardiac fibroblasts and the results were directly tested by MTT assay. c Cardiac fibroblasts cells were transfected with 60 nM miR-21 inhibitor for 48 h. Then cells were harvested and the cell-cycle distribution was analyzed by Flow cytometry analysis. Data are representative of at least three separate experiments. *P < 0.05, **P < 0.01 versus Vehicle or Control; #P < 0.05 versus modelCao et al. BMC Cardiovascular Disorders (2017) 17:Page 8 ofABCDFig. 5 miR-21 negative control CADM1 expression. a Real-time PCR showed that transfection miR-21 mimic decreased the expression of CADM1 mRNA in cardiac fibroblasts cells. miR-21 mimic group is decrea.
