a West China School of Public Health and Healthy Food Evaluation Center, Sichuan University , Chengdu , China.
b Research Center for Occupational Respiratory Diseases, West China Fourth Hospital, Sichuan University , Chengdu , China.
Pharm Biol. 2019 Dec;57(1):470-476. doi: 10.1080/13880209.2019.1640747.
We reported that D-4F, an apolipoprotein A-I (Apo A-I) mimetic polypeptide with 18 d-amino acids, suppressed IL-4 induced macrophage alternative activation and TGF-β1 expression in phorbol 12-myristate 13-acetate (PMA) treated human acute monocytic leukemia cells (THP-1). Macrophage alternative activation, TGF-β1 and epithelial-mesenchymal transition (EMT) are intensively involved in pulmonary fibrosis. Recent studies demonstrated that Apo A-I resolved established pulmonary fibrotic nodules, and D-4F inhibited TGF-β1 induced EMT in alveolar cells. Therefore, this study evaluated the effects of D-4F on IL-4 induced macrophage alternative activation and TGF-β1 expression. THP-1 cells were simulated with PMA (100 ng/mL) for 48 h and treated with medium control, IL-4 (20 ng/mL) alone, or IL-4 (20 ng/mL) in the presence of D-4F (1, 5, and 10 μg/mL) for 24 and 48 h. Flow cytometry, RT-PCR and ELISA evaluations were performed to investigate the subsequent effects of D-4F. Compared to stimulation with IL-4 alone, 1, 5, and 10 μg/mL of D-4F reduced alternative activation by 45.38%, 59.98%, and 60.10%, increased TNF-α mRNA levels by 8%, 11%, and 16% and decreased TGF-β1 mRNA levels by 21%, 37%, and 39%, respectively (all ≤ 0.05). In addition, TNF-α protein levels increased from 388 pg/mL (IL-4 alone) to 429, 475, and 487 pg/mL (1, 5, and 10 μg/mL D-4F), while TGF-β1 protein levels dropped from 27.01 pg/mL (IL-4 alone) to 19.15, 12.27, and 10.47 pg/mL (1, 5, and 10 μg/mL D-4F). D-4F suppressed IL-4 induced macrophage alternative activation and pro-fibrotic TGF-β1 expression.
我们曾报道，D-4F 是一种载脂蛋白 A-I（Apo A-I）模拟多肽，具有 18 个 D-氨基酸，可抑制佛波醇 12-肉豆蔻酸 13-乙酸酯（PMA）处理的人急性单核细胞白血病细胞（THP-1）中白细胞介素 4（IL-4）诱导的巨噬细胞替代激活和转化生长因子-β1（TGF-β1）表达。巨噬细胞替代激活、TGF-β1 和上皮-间充质转化（EMT）在肺纤维化中广泛参与。最近的研究表明，Apo A-I 可解决已建立的肺纤维性结节，D-4F 可抑制肺泡细胞中 TGF-β1 诱导的 EMT。因此，本研究评估了 D-4F 对 IL-4 诱导的巨噬细胞替代激活和 TGF-β1 表达的影响。THP-1 细胞用 PMA（100ng/mL）模拟 48h，并用培养基对照、IL-4（20ng/mL）单独或 IL-4（20ng/mL）加 D-4F（1、5 和 10μg/mL）处理 24 和 48h。流式细胞术、RT-PCR 和 ELISA 评估用于研究 D-4F 的后续作用。与单独用 IL-4 刺激相比，1、5 和 10μg/mL 的 D-4F 分别降低了 45.38%、59.98%和 60.10%的替代激活，增加了 8%、11%和 16%的 TNF-α mRNA 水平，降低了 21%、37%和 39%的 TGF-β1 mRNA 水平（均 ≤ 0.05）。此外，TNF-α 蛋白水平从单独用 IL-4 的 388pg/mL 增加到 429、475 和 487pg/mL（1、5 和 10μg/mL D-4F），而 TGF-β1 蛋白水平从单独用 IL-4 的 27.01pg/mL 降低至 19.15、12.27 和 10.47pg/mL（1、5 和 10μg/mL D-4F）。D-4F 抑制了 IL-4 诱导的巨噬细胞替代激活和促纤维化 TGF-β1 表达。
