This study deciphered the ameliorating effect and molecular mechanism of the total glucosides of White Paeony Capsules(TGP)in the treatment of mice model with acute lung injury(ALI)via NOD-like receptor thermal protein domain associated protein 3(NLRP3)signaling pathway of the inflammasome.The study established an inflammasome activation model of primed bone marrow-de-rived macrophages(BMDMs),and its molecular mechanism was investigated by Western blot(WB),immunofluorescence staining,enzyme-linked immunosorbent assay(ELISA),and flow cytometry.C57BL/6J mice were randomly divided into a blank control group,a TGP group,a model group(LPS group),LPS+low-and high-dose TGP groups,LPS+MCC950 group,and LPS+MCC950+TGP group,with eight mice per group.The ALI model was induced in mice.Finally,bronchoalveolar lavage fluid(BALF)and lung tissue were collected.Lung index and lung weight wet-to-dry ratio were determined for each group of mice.The pathological changes in lung tissue were observed through hematoxylin-eosin(HE)staining.The number of neutrophils in the BALF of each group was detected using flow cytometry.The levels of interleukin(IL)-1β,IL-6,and tumor necrosis factor(TNF)-α in the BALF were determined by ELISA.The expressions of IL-1β,IL-18,IL-6,and TNF-α in the lung tissue were determined by real-time quantitative PCR(RT-qPCR).This study demonstrated that TGP dramatically blocked the activation of the NLRP3 inflammasome by inhibiting the production of upstream mitochondrial reactive oxygen species(mtROS)and the subsequent oligomerization of apoptosis-associated specks(ASC).Additionally,in the ALI mice model,compared with the blank control group,the model group showed alveolar structure rupture,thic-kening of alveolar septa,and dramatically increased lung index,lung weight wet-to-dry ratio in lung tissue,neutrophil count,and in-flammatory factor levels.Compared with the model group,the pathological morphology of lung tissue was significantly ameliorated in the TGP and MCC950 groups,and the lung index and lung weight wet-to-dry ratio were significantly reduced.Neutrophil counts were re-duced,and levels of inflammatory factors were significantly downregulated.Notably,compared with the MCC950 group,there was no significant difference in effect in the MCC950+TGP group.Collectively,the study reveals that TGP may ameliorate ALI in mice by in-hibiting the activation of NLRP3 inflammasome,providing a safe and effective drug candidate for the prevention or treatment of ALI/ARDS.
total glucosides of White Paeony CapsulesNLRP3 inflammasomemitochondrial reactive oxygen speciesASC oli-gomerizationLPS-induced acute lung injury
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