背景:特发性肺纤维化(IPF)的治疗由于其持续的进展仍然具有挑战性,严峻的预后,以及有效治疗方案的稀缺。新兴证据强烈支持肺泡上皮细胞(AECs)加速衰老在驱动IPF进展中的关键作用。因此,靶向衰老AECs是IPF的一种有前途的治疗策略。
目的:姜黄素类似物EF24是姜黄素的衍生物,显示出增强的生物活性,包括抗炎,抗肿瘤和抗衰老特性。这项研究的目的是阐明EF24在IPF治疗中的治疗潜力和潜在的分子机制。
方法:使用博来霉素诱导A549和ATII细胞衰老。使用不同方法检查衰老标志物,包括衰老相关β-半乳糖苷酶(SA-β-gal)染色,西方印迹,和q-PCR。对小鼠气管内施用博来霉素以诱导肺纤维化。这通过显微计算机断层扫描(CT)得到了验证,马尾松三色染色,和透射电子显微镜(TEM)。通过评估PTEN的表达,在体外和体内确定EF24在IPF中的作用和潜在机制。AKT/mTOR/NF-κB信号通路,使用蛋白质印迹或流式细胞术进行线粒体自噬。
结果:我们确定姜黄素类似物EF24是抗IPF的12种化合物中最有希望的候选物。EF24治疗显着减少博来霉素诱导的衰老AECs中的衰老生物标志物,包括SA-β-Gal,PAI-1、P21和衰老相关分泌表型(SASP)。EF24还有效抑制由衰老AECs或TGF-β诱导的成纤维细胞活化。我们发现,PTEN激活是EF24通过抑制AKT/mTOR/NF-κB信号通路抑制AECs衰老的重要因素。此外,EF24通过诱导线粒体自噬改善线粒体功能障碍。此外,EF24给药可显着降低博来霉素在小鼠肺组织中诱导的衰老表型。值得注意的是,EF24减轻纤维化并促进IPF急性和慢性阶段的整体健康益处。提示其在IPF治疗中的治疗潜力。
结论:这些研究结果共同强调了EF24作为一种新的有效治疗剂,通过抑制AECs的衰老来对抗IPF。
BACKGROUND: Treating Idiopathic pulmonary fibrosis (IPF) remains challenging owing to its relentless progression, grim prognosis, and the scarcity of effective treatment options. Emerging evidence strongly supports the critical role of accelerated senescence in alveolar epithelial cells (AECs) in driving the progression of IPF. Consequently, targeting senescent AECs emerges as a promising therapeutic strategy for IPF.
OBJECTIVE: Curcumin analogue EF24 is a derivative of curcumin and shows heightened bioactivity encompassing anti-inflammatory, anti-tumor and anti-aging properties. The objective of this study was to elucidate the therapeutic potential and underlying molecular mechanisms of EF24 in the treatment of IPF.
METHODS: A549 and ATII cells were induced to become senescent using bleomycin. Senescence markers were examined using different methods including senescence-associated β-galactosidase (SA-β-gal) staining, western blotting, and q-PCR. Mice were intratracheally administrated with bleomycin to induce pulmonary fibrosis. This was validated by micro-computed tomography (CT), masson trichrome staining, and transmission electron microscope (TEM). The role and underlying mechanisms of EF24 in IPF were determined in vitro and in vivo by evaluating the expressions of
PTEN, AKT/mTOR/NF-κB signaling pathway, and mitophagy using western blotting or flow cytometry.
RESULTS: We identified that the curcumin analogue EF24 was the most promising candidate among 12 compounds against IPF. EF24 treatment significantly reduced senescence biomarkers in bleomycin-induced senescent AECs, including SA-β-Gal, PAI-1, P21, and the senescence-associated secretory phenotype (SASP). EF24 also effectively inhibited fibroblast activation which was induced by senescent AECs or TGF-β. We revealed that
PTEN activation was integral for EF24 to inhibit AECs senescence by suppressing the AKT/mTOR/NF-κB signaling pathway. Additionally, EF24 improved mitochondrial dysfunction through induction of mitophagy. Furthermore, EF24 administration significantly reduced the senescent phenotype induced by bleomycin in the lung tissues of mice. Notably, EF24 mitigates fibrosis and promotes overall health benefits in both the acute and chronic phases of IPF, suggesting its therapeutic potential in IPF treatment.
CONCLUSIONS: These findings collectively highlight EF24 as a new and effective therapeutic agent against IPF by inhibiting senescence in AECs.