Madeng’ai Ethyl Acetate Extract Attenuates Lipopolysaccharide-Induced Inflammation and Acute Lung Injury

Madeng’ai, the dried root of Potentilla freyniana Bornm., is used in Dong medicine for infections and wound care. This study examined the chemical composition and prophylactic protective effects of the ethyl acetate fraction of its ethanolic root extract (referred to as MEA) in lipopolysaccharide (LPS)-induced acute lung injury (ALI). Ultra-high-performance liquid chromatography coupled to Q-Exactive Orbitrap mass spectrometry detected 77 constituents, comprising 13 compounds authenticated with reference standards and 64 constituents tentatively annotated from accurate mass and tandem mass spectrometry (MS/MS) fragmentation. Network pharmacology prioritized six candidate compounds, 120 overlapping targets, and nine hub targets, and molecular docking explored theoretical interactions with inflammation-related proteins. MEA was evaluated in LPS-stimulated MH-S alveolar macrophages and in male C57BL/6J mice receiving oral pretreatment for seven days prior to intraperitoneal LPS challenge. In MH-S cells, MEA pretreatment reduced nitrite, interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA)-detectable intracellular oxidative fluorescence, and Nlrp3, Pycard, Casp1, and Gsdmd mRNA levels. In mice, seven-day oral pretreatment with MEA attenuated qualitative histological lung injury, pulmonary edema, and serum cytokine elevation, accompanied by lower lung protein levels of NOD-like receptor family pyrin domain-containing 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), pro-caspase-1, and the ~30 kDa N-terminal fragment of gasdermin D (GSDMD-N), together with related transcripts. These findings indicate that prophylactic MEA administration attenuates LPS-induced macrophage inflammation and experimental lung injury, accompanied by reduced intracellular oxidative stress and decreased expression of NLRP3–GSDMD pathway-associated markers.