Table 7_Resveratrol attenuates pulmonary fibrosis by inhibiting alveolar epithelial senescence via targeting SASP-related proteins: an integrated bioinformatics-experimental study.pdf

BackgroundPulmonary fibrosis (PF) is a progressive and fatal interstitial lung disease with limited treatment options. Premature senescence of alveolar epithelial type II cells (AT2 cells) plays a critical role in PF pathogenesis. This study aimed to identify natural compounds targeting senescence-related pathways for PF treatment.
MethodsAn integrated approach was implemented, combining bioinformatics, artificial intelligence (AI)-assisted molecular docking, ADMET (absorption, distribution, metabolism, excretion, and toxicity) profiling, and experimental validation. Core targets associated with aging-related pulmonary fibrosis (PF) were identified via database mining (GeneCards and AgingAtlas) and protein-protein interaction (PPI) network analysis. Natural compounds were screened using the HERB database, and resveratrol (RES) was selected due to its multi-target activity and favorable ADMET characteristics. The efficacy of RES was evaluated through in vitro experiments using bleomycin (BLM)-induced senescent A549 alveolar epithelial cells and in vivo studies in a BLM-induced PF mouse model (C57BL/6J). Molecular docking simulations were performed to predict the binding affinity between RES and key targets, including SERPINE1, MMP2, and IL-6.
ResultsBioinformatics identified 322 aging-related PF targets, with TP53, AKT1, STAT3, JUN, and NFKB1 as core regulators. Resveratrol was selected as a top candidate modulating all five core targets and exhibiting optimal drug-likeness. Molecular docking and dynamics simulations confirmed strong binding affinity between RES and key senescence-associated proteins (SERPINE1: −8 kcal/mol; MMP2: −7.5 kcal/mol; IL-6: −7.1 kcal/mol). In vitro, RES (10–40 μM) significantly suppressed bleomycin-induced senescence in A549 cells, reducing SA-β-Gal activity and downregulating SERPINE1, MMP2, and IL6 expression. In vivo, RES treatment (20–80 mg/kg, 21 days) attenuated bleomycin-induced PF in mice, improving weight loss, reducing alveolar damage, inflammation, and collagen deposition (Masson’s trichrome) in a dose-dependent manner.
ConclusionResveratrol effectively inhibits alveolar epithelial cell senescence and ameliorates pulmonary fibrosis, likely by targeting key senescence-associated pathways (e.g., SERPINE1, MMP2, IL-6). This study provides a promising transdisciplinary strategy for anti-fibrotic drug discovery and highlights RES as a potential therapeutic candidate for PF.

【背景】肺纤维化（Pulmonary fibrosis, PF）是一种进展性致死性间质性肺疾病，临床治疗手段有限。肺泡II型上皮细胞（alveolar epithelial type II cells, AT2 cells）的过早衰老在PF的发病机制中发挥关键作用。本研究旨在筛选靶向衰老相关通路的天然化合物，用于PF治疗。【方法】本研究采用整合研究策略，结合生物信息学（bioinformatics）、人工智能（AI）辅助分子对接、ADMET（吸收、分布、代谢、排泄及毒性）谱分析及实验验证。通过GeneCards（基因卡）与AgingAtlas（衰老图谱）数据库挖掘，以及蛋白质相互作用（protein-protein interaction, PPI）网络分析，筛选与衰老相关肺纤维化相关的核心靶点。借助HERB数据库筛选天然化合物，最终选择白藜芦醇（resveratrol, RES），因其具备多靶点活性且ADMET特性优良。通过博来霉素（bleomycin, BLM）诱导的衰老A549肺泡上皮细胞的体外实验，以及博来霉素诱导的PF小鼠模型（C57BL/6J）的体内实验，评估RES的治疗疗效。通过分子对接模拟（molecular docking simulations），预测RES与SERPINE1、MMP2、IL-6等关键靶点的结合亲和力（binding affinity）。【结果】生物信息学分析共筛选出322个衰老相关PF靶点，其中TP53、AKT1、STAT3、JUN及NFKB1为核心调控因子。白藜芦醇可调控全部5个核心靶点，且具备最优的药物相似性（drug-likeness），被选为最优候选化合物。分子对接及动力学模拟（molecular dynamics simulations）证实，RES与关键衰老相关蛋白具有较强结合亲和力（SERPINE1：−8 kcal/mol；MMP2：−7.5 kcal/mol；IL-6：−7.1 kcal/mol）。体外实验中，10~40 μM的RES可显著抑制博来霉素诱导的A549细胞衰老，降低SA-β-Gal（衰老相关β-半乳糖苷酶）活性，并下调SERPINE1、MMP2及IL6的表达。体内实验中，20~80 mg/kg的RES连续给药21天，可剂量依赖性减轻博来霉素诱导的小鼠PF，改善体重减轻情况，减轻肺泡损伤、炎症反应及胶原沉积（经Masson三色染色Masson’s trichrome验证）。【结论】白藜芦醇可有效抑制肺泡上皮细胞衰老并改善肺纤维化，其作用机制可能通过靶向关键衰老相关通路（如SERPINE1、MMP2、IL-6）实现。本研究为抗纤维化药物发现（anti-fibrotic drug discovery）提供了极具前景的跨学科策略（transdisciplinary strategy），并证实RES是治疗PF的潜在候选药物。