Bleomycin (BLM) injury is associated with the severity of acute lung injury (ALI) leading to fibrosis, a high-morbidity, and high-mortality respiratory disease of unknown etiology. BLM-induced ALI is marked by the activation of a potent fibrogenic cytokine transcription growth factor beta-1 (TGFβ-1), which is considered a critical cytokine in the progression of alveolar injury. Previously, our work demonstrated that a diet-derived compound curcumin (diferuloylmethane), represents its antioxidative and antifibrotic application in TGF-β1-mediated BLM-induced alveolar basal epithelial cells. However, curcumin-specific protein targets, as well as its mechanism using mass spectrometry-based proteomic approach, remain elusive. To elucidate the underlying mechanism, a quantitative proteomics approach and bioinformatics analysis were employed to identify the protein targets of curcumin in BLM or TGF-β1-treated cells. With subsequent in vitro experiments, curcumin-related pathways and cellular processes were predicted and validated. The current study discusses two separate proteomics experiments using BLM and TGF-β1-treated cells with the proteomics approach, various unique target proteins were identified, and proteomic analysis revealed that curcumin reversed the expressions of unique proteins like DNA topoisomerase 2-alpha (TOP2A), kinesin-like protein (KIF11), centromere protein F (CENPF), and so on BLM or TGF-β1 injury. For the first time, the current study reveals that curcumin restores TGF-β1 induced peroxisomes like PEX-13, PEX-14, PEX-19, and ACOX1. This was verified by subsequent in vitro assays. This study generated molecular evidence to deepen our understanding of the therapeutic role of curcumin at the proteomic level and may be useful to identify molecular targets for future drug discovery.

中文翻译：

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姜黄素在急性肺损伤体外模型中的独特分子靶点与抗氧化和抗纤维化活性相关：蛋白质组学方法

博来霉素 (BLM) 损伤与导致纤维化的急性肺损伤 (ALI) 的严重程度相关，这是一种病因不明的高发病率和高死亡率呼吸道疾病。BLM 诱导的 ALI 以激活有效的纤维化细胞因子转录生长因子 β-1 (TGFβ-1) 为标志，TGFβ-1 被认为是肺泡损伤进展中的关键细胞因子。此前，我们的工作表明饮食衍生的化合物姜黄素（二乙酰甲烷）代表其在 TGF-β1 介导的 BLM 诱导的肺泡基底上皮细胞中的抗氧化和抗纤维化应用。然而，姜黄素特异性蛋白质靶标及其使用基于质谱的蛋白质组学方法的机制仍然难以捉摸。为了阐明潜在的机制，采用定量蛋白质组学方法和生物信息学分析来鉴定 BLM 或 TGF-β1 处理的细胞中姜黄素的蛋白质靶标。通过随后的体外实验，预测和验证了姜黄素相关途径和细胞过程。目前的研究使用蛋白质组学方法讨论了使用 BLM 和 TGF-β1 处理的细胞的两个单独的蛋白质组学实验，鉴定了各种独特的靶蛋白，蛋白质组学分析显示姜黄素逆转了独特蛋白质的表达，如 DNA 拓扑异构酶 2-α (TOP2A) 、驱动蛋白样蛋白（KIF11）、着丝粒蛋白F（CENPF）等BLM或TGF-β1损伤。目前的研究首次揭示姜黄素可恢复 TGF-β1 诱导的过氧化物酶体，如 PEX-13、PEX-14、PEX-19 和 ACOX1。这通过随后的体外测定得到证实。