Department of Integrated Cardiology, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang 830000, P.R. China.
School of Health Management, Xinjiang Medical University, Urumqi, Xinjiang 830054, P.R. China.
Mol Med Rep. 2024 Nov;30(5). doi: 10.3892/mmr.2024.13333. Epub 2024 Sep 20.
Fibrosis is the basis of structural remodeling in atrial fibrillation (AF), during which inflammation is crucial. Programmed cell death factor 4 (PDCD4) is a newly identified inflammatory gene, with unknown mechanisms of action in AF. The present study aimed to elucidate the effects of PDCD4 on the inflammation and structural remodeling of atrial myocytes. For this purpose, a PDCD4 overexpression plasmid (oePDCD4) and PDCD4 small interfering (si)RNA (siPDCD4) were used to modulate PDCD4 expression in mouse atrial myocytes (HL‑1 cells). The expression of PDCD4 was detected using reverse transcription‑quantitative PCR and western blot analysis. The optimal drug concentrations of peroxisome proliferator‑activated receptor γ (PPARγ) agonist (pioglitazone hydrochloride), NF‑κB inhibitor (CBL0137), PPARγ inhibitor (GW9962) and NF‑κB agonist (betulinic acid) were screened using a Cell Counting Kit‑8 assay. The levels of inflammatory factors were detected using enzyme‑linked immunosorbent assays, the expression levels of fibrosis‑related proteins and NF‑κB subunits were detected using western blot analysis, and the expression of phosphorylated (p‑)p65/p65 was detected using immunofluorescence staining. The results revealed that PDCD4 overexpression increased the levels of fibrotic factors (collagen I, collagen III, fibronectin, α‑smooth muscle actin and matrix metalloproteinase 2), pro‑inflammatory cytokines (IFN‑γ, IL‑6, IL‑17A and TNF‑α) and p‑p65, whereas it reduced the levels of anti‑inflammatory cytokines (IL‑4) in HL‑1 cells. Additionally, treatment with the PPARγ agonist and NF‑κB inhibitor reversed the levels of fibrotic‑, pro‑inflammatory and anti‑inflammatory factors in oePDCD4‑HL‑1 cells. By contrast, PDCD4 silencing exerted the opposite effects on fibrotic factors, pro‑inflammatory cytokines, anti‑inflammatory cytokines and p‑p65. In addition, treatment with the PPARγ inhibitor and NF‑κB agonist reversed the levels of fibrotic‑, pro‑inflammatory and anti‑inflammatory factors in siPDCD4‑HL‑1 cells. In conclusion, the present study demonstrated that PDCD4 may induce inflammation and fibrosis by activating the PPARγ/NF‑κB signaling pathway, thereby promoting the structural remodeling of atrial myocytes in AF.
纤维化是心房颤动 (AF) 结构重构的基础,在此过程中炎症至关重要。程序性细胞死亡因子 4 (PDCD4) 是一种新发现的炎症基因,其在 AF 中的作用机制尚不清楚。本研究旨在阐明 PDCD4 对心房肌细胞炎症和结构重构的影响。为此,使用 PDCD4 过表达质粒 (oePDCD4) 和 PDCD4 小干扰 (si)RNA (siPDCD4) 调节小鼠心房肌细胞 (HL-1 细胞) 中的 PDCD4 表达。使用逆转录-定量 PCR 和 Western blot 分析检测 PDCD4 的表达。使用细胞计数试剂盒-8 测定筛选过氧化物酶体增殖物激活受体 γ (PPARγ) 激动剂 (盐酸吡格列酮)、NF-κB 抑制剂 (CBL0137)、PPARγ 抑制剂 (GW9962) 和 NF-κB 激动剂 (白桦脂酸) 的最佳药物浓度。使用酶联免疫吸附试验检测炎症因子水平,使用 Western blot 分析检测纤维化相关蛋白和 NF-κB 亚基的表达水平,使用免疫荧光染色检测磷酸化 (p)-p65/p65 的表达。结果显示,PDCD4 过表达增加了纤维化因子 (胶原 I、胶原 III、纤连蛋白、α-平滑肌肌动蛋白和基质金属蛋白酶 2)、促炎细胞因子 (IFN-γ、IL-6、IL-17A 和 TNF-α) 和 p-p65 的水平,而降低了 HL-1 细胞中抗炎细胞因子 (IL-4) 的水平。此外,PPARγ 激动剂和 NF-κB 抑制剂处理逆转了 oePDCD4-HL-1 细胞中纤维化、促炎和抗炎因子的水平。相比之下,PDCD4 沉默对纤维化因子、促炎细胞因子、抗炎细胞因子和 p-p65 产生相反的影响。此外,PPARγ 抑制剂和 NF-κB 激动剂处理逆转了 siPDCD4-HL-1 细胞中纤维化、促炎和抗炎因子的水平。综上所述,本研究表明,PDCD4 可能通过激活 PPARγ/NF-κB 信号通路诱导炎症和纤维化,从而促进 AF 中心房肌细胞的结构重构。