大成奇汤散剂颗粒通过抑制PANoptosis改善LPS诱导的急性肺损伤（ALI） in vivo和in vitro

急性肺损伤（ALI）是一种严重威胁健康的疾病，表现为肺部强烈的炎症反应，若不及时治疗可发展为急性呼吸窘迫综合征（ARDS）。大成奇汤散剂颗粒（DDG）在肺部具有保护作用，但其潜在调节机制以缓解ALI的作用尚需深入研究。本研究旨在探讨DDG在体内外LPS诱导的ALI模型中的作用及其潜在机制。采用LPS处理的Balb/c小鼠和BEAS-2B细胞分别建立体内外ALI模型。通过苏木精-伊红（HE）染色、肺组织的湿/干比（W/D）计算，以及BALF中总蛋白和乳酸脱氢酶（LDH）水平检测，评估肺组织损伤和肺水肿程度。采用酶联免疫吸附试验（ELISA）检测BALF、血清和细胞上清液中的肿瘤坏死因子α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-18（IL-18）水平。qRT-PCR检测肺组织和BEAS-2B细胞中炎症因子、Z-DNA结合蛋白1（ZBP1）及受体相互作用蛋白激酶1（RIPK1）的表达。免疫双标染色和免疫共沉淀技术用于检测ZBP1与RIPK1的表达及共定位。Cell Counting Kit-8（CCK-8）检测LPS和DDG对BEAS-2B细胞活性的影响。Western blot（WB）分析肺组织和BEAS-2B细胞中PANoptosis相关蛋白的表达水平。结果显示，DDG预处理能剂量依赖性改善ALI小鼠肺组织的病理变化，降低肺W/D比例、总蛋白浓度及BALF中的LDH含量。在体外，DDG逆转了LPS对BEAS-2B细胞活力的抑制作用，同时显著降低了炎症因子水平。进一步发现，DDG可以抑制PANoptosis相关蛋白的表达，特别是上游关键调控分子ZBP1和RIPK1。总之，DDG通过抑制过度炎症和PANoptosis缓解LPS诱导的ALI，表现出良好的抗炎和肺保护作用。本研究为DDG的进一步开发提供理论基础，并为靶向PANoptosis的ALI治疗开辟了新前景。

Acute lung injury (ALI) is a serious health-threatening syndrome of intense inflammatory response in the lungs, with progression leading to acute respiratory distress syndrome (ARDS). Dachengqi decoction dispensing granule (DDG) has a pulmonary protective role, but its potential modulatory mechanism to alleviate ALI needs further excavation.This study aims to investigate the effect and potential mechanism of DDG on lipopolysaccharide (LPS)-induced ALI models in vivo and in vitro.LPS-treated Balb/c mice and BEAS-2B cells were used to construct in vivo and in vitro ALI models, respectively. Hematoxylin-eosin (HE), Wet weight/Dry weight (W/D) calculation of lung tissue, and total protein and Lactic dehydrogenase (LDH) assays in BALF were performed to assess the extent of lung tissue injury and pulmonary edema. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of tumor necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β), and interleukin-18 (IL-18) in BALF, serum, and cell supernatant. The qRT-PCR was used to detect inflammatory factors, Z-DNA binding protein 1 (ZBP1), and receptor-interacting protein kinase 1 (RIPK1) expression in lung tissues and BEAS-2B cells. Double immunofluorescence staining and co-immunoprecipitation were used to detect the relative expression and co-localization of ZBP1 and RIPK1. The effects of LPS and DDG on BEAS-2B cell activity were detected by Cell Counting Kit-8 (CCK-8). Western blot (WB) was performed to analyze the expression of PANoptosis-related proteins in lung tissues and BEAS-2B cells.In vivo, DDG pretreatment could dose-dependently improve the pathological changes of lung tissue in ALI mice, and reduce the W/D ratio of lung, total protein concentration, and LDH content in BALF. In vitro, DDG reversed the inhibitory effect of LPS on BEAS-2B cell viability. Meanwhile, DDG significantly reduced the levels of inflammatory factors in vitro and in vivo. In addition, DDG could inhibit the expression levels of PANoptosis-related proteins, especially the upstream key regulatory molecules ZBP1 and RIPK1.DDG could inhibit excessive inflammation and PANoptosis to alleviate LPS-induced ALI, thus possessing good anti-inflammatory and lung-protective effects. This study establishes a theoretical basis for the further development of DDG and provides a new prospect for ALI treatment by targeting PANoptosis.