Chen Jian, Li Fu, Luo Wang-Sheng, Zhu Mei-Fang, Zhao Neng-Jiang, Zhang Zhi-Hai, Chen Ya-Feng, Feng Dian-Xu, Yang Shu-Yu, Sun Wen-Jie
Institute of Vascular Anomalies, Shanghai TCM-Integrated Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200082, China; Department of Public Health, International College, Krirk University, Bangkok, Thailand.
Department of Hepatopancreatobiliary Surgery, Shuguang Hospital affliated to Shanghai University of Traditional Chinese Medincine, Shanghai 201203, China.
Cell Signal. 2025 Jul;131:111733. doi: 10.1016/j.cellsig.2025.111733. Epub 2025 Mar 11.
This study aimed to elucidate the protective effects of Da Cheng Qi Decoction (DCQD) on severe acute pancreatitis (SAP) by targeting ferroptosis in pancreatic acinar cells and to establish a predictive signature and nomogram for acute pancreatitis (AP) risk assessment.
We utilized microarray analysis to delineate gene expression patterns among 32 healthy controls and 87 AP patients stratified by severity. Employing SAP models and NOX2-deficient cells, we investigated the molecular underpinnings of ferroptosis. The impact of DCQD and the ferroptosis inhibitor Fer-1 on gene expression, oxidative stress, and inflammation was assessed. Machine learning algorithms identified differentially expressed genes (DEGs) sensitive to DCQD, SAP, and ferroptosis (DSNFGs), which were validated across multiple datasets. A predictive nomogram integrating DSNFGs was developed, and single-cell analysis provided a comprehensive view of the cellular dynamics.
The microarray analysis revealed upregulation of NOX2 and downregulation of GPX4 in AP, with expression patterns correlating with disease severity. DCQD ameliorated SAP-induced pancreatic acinar cell damage and ferroptosis by reducing inflammatory markers and enhancing GPX4 expression. NOX2 knockout mitigated ferroptosis in SAP models, suggesting a key role in the disease process. DCQD and Fer-1 differentially regulated the expression of ferroptosis-related genes, reduced reactive oxygen species (ROS) and high-mobility group box 1 (HMGB1) levels, and suppressed the inflammatory response in a SAP mouse model. The HPLC analysis of DCQD constituents indicated eight components (aloe-emodin, rhein, emodin, chrysophanol, naringin, hesperidin, magnolol, and honokiol) with the capacity to modulate ferroptosis. Venn analysis identified 48 DSNFGs, with a subset of five genes demonstrating significant predictive value. The developed nomogram, based on LASSO regression, showed high accuracy in validation cohorts. Single-cell RNA sequencing (scRNA-seq) and CellChat analysis uncovered heterogeneity and cell-cell communication networks in the pancreas during recovery from pancreatitis, implicating several signaling pathways.
DCQD and its eight ingredients exert its protective effect in SAP by inhibiting ferroptosis through the NOX2/GPX4 pathway. The DCQD-SAP-ferroptosis-related signature and nomogram offer a novel tool for AP risk assessment, prognosis prediction, and personalized therapeutic strategies in SAP management.
本研究旨在通过靶向胰腺腺泡细胞铁死亡阐明大承气汤（DCQD）对重症急性胰腺炎（SAP）的保护作用，并建立用于急性胰腺炎（AP）风险评估的预测特征和列线图。
我们利用微阵列分析来描绘32名健康对照者和87名按严重程度分层的AP患者之间的基因表达模式。采用SAP模型和NOX2缺陷细胞，我们研究了铁死亡的分子机制。评估了DCQD和铁死亡抑制剂Fer-1对基因表达、氧化应激和炎症的影响。机器学习算法鉴定出对DCQD、SAP和铁死亡敏感的差异表达基因（DSNFGs），并在多个数据集中进行了验证。开发了一个整合DSNFGs的预测列线图，单细胞分析提供了细胞动力学的全面视图。
微阵列分析显示AP中NOX2上调而GPX4下调，并与疾病严重程度相关。DCQD通过减少炎症标志物和增强GPX4表达改善了SAP诱导的胰腺腺泡细胞损伤和铁死亡。NOX2基因敲除减轻了SAP模型中的铁死亡，表明其在疾病过程中起关键作用。DCQD和Fer-1对铁死亡相关基因的表达有不同调节作用，降低了活性氧（ROS）和高迁移率族蛋白B1（HMGB1）水平，并在SAP小鼠模型中抑制了炎症反应。DCQD成分的高效液相色谱分析表明有八种成分（芦荟大黄素、大黄酸、大黄素、 Chrysophanol、柚皮苷、橙皮苷、厚朴酚和和厚朴酚）具有调节铁死亡的能力。维恩分析鉴定出48个DSNFGs，其中五个基因的子集显示出显著的预测价值。基于LASSO回归开发的列线图在验证队列中显示出高准确性。单细胞RNA测序（scRNA-seq）和CellChat分析揭示了胰腺炎恢复过程中胰腺的异质性和细胞间通讯网络，涉及多个信号通路。
DCQD及其八种成分通过NOX2/GPX4途径抑制铁死亡，在SAP中发挥保护作用。DCQD-SAP-铁死亡相关特征和列线图为AP风险评估、预后预测以及SAP管理中的个性化治疗策略提供了一种新工具。
