调控miRNA-182-5p表达对AC16细胞纤维化的影响

Regulatory Effects of miRNA-182-5p Expression on Fibrosis in AC16 Cells

摘要: 目的：探讨miR-182-5p在转化生长因子β1 (TGF-β1)诱导的AC16人心肌细胞纤维化中的调控作用及其分子机制。方法：以TGF-β1处理AC16细胞构建心肌纤维化模型，通过CCK-8法检测不同处理条件下细胞活力，筛选最佳诱导浓度和时间。实验设正常对照组、TGF-β1组、miR-182-5p过表达组、miR-182-5p抑制组及miRNA阴性对照(NC)组。采用Lipofectamine^TM 3000进行细胞转染。通过实时定量PCR (qPCR)、Western blot及免疫荧光检测纤维化标志物α-SMA、Collagen I及Collagen III的表达水平。结果：与NC组相比，miR-182-5p过表达组α-SMA、Collagen I和Collagen III的mRNA及蛋白表达水平均显著降低(P < 0.05)，而抑制miR-182-5p则显著升高上述指标(P < 0.05)。NC组与TGF-β1组相比差异无统计学意义。结论：miR-182-5p可抑制TGF-β1诱导的AC16细胞纤维化反应，提示其可能成为心肌纤维化干预的潜在分子靶点。

Abstract: Objective: To investigate the regulatory role of miR-182-5p in transforming growth factor-β1 (TGF-β1)-induced fibrosis in AC16 human cardiomyocytes and explore its underlying molecular mechanisms. Methods: A myocardial fibrosis model was established by treating AC16 cells with TGF-β1. Cell viability was assessed using the CCK-8 assay to determine the optimal induction concentration and duration. The experiment included five groups: control group, TGF-β1 group, miR-182-5p overexpression group, miR-182-5p inhibition group, and miRNA negative control (NC) group. Transfection was performed using Lipofectamine^TM 3000. The expression levels of fibrosis-related markers, including α-SMA, collagen I, and collagen III, were evaluated by quantitative PCR (qPCR), Western blotting, and immunofluorescence. Results: Compared with the NC group, the miR-182-5p overexpression group showed significantly reduced mRNA and protein expression levels of α-SMA, collagen I, and collagen III (P < 0.05), whereas inhibition of miR-182-5p significantly increased their expression (P < 0.05). No significant difference was observed between the NC group and the TGF-β1 group. Conclusion: miR-182-5p inhibits TGF-β1-induced fibrotic responses in AC16 cells and may serve as a potential therapeutic target for myocardial fibrosis.