Inflammation plays an important role in the development of acute lung injury (ALI). Severe pulmonary inflammation can cause acute respiratory distress syndrome (ARDS) or even death. Expression of proinflammatory interleukin-1β(IL-1β) and inducible nitric oxide synthase (iNOS) in the process of pulmonary inflammation will further exacerbate the severity of ALI. The purpose of this study was to explore the effect of Palrnatine (Pa) on lipopolysaccharide (LPS)-induced mouse ALI and its underlying mechanism. Pa, a natural product, has a wide range of pharmacological activities with the potential to protect against lung injury. Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) assays were performed to detect the expression and translation of inflammatory genes and proteins in vitro and in vivo. Immunoprecipitation was used to detect the degree of P65 translocation into the nucleus. We also used molecular modeling to further clarify the mechanism of action. The results showed that Pa pretreatment could significantly inhibit the expression and secretion of the inflammatory cytokine IL-1β, and significantly reduce the protein level of the proinflammatory protease iNOS, in both in vivo and in vitro models induced by LPS. Further mechanism studies showed that Pa could significantly inhibit the activation of the protein kinase B (Akt)/nuclear factor-κB (NF-κB) signaling pathway in the LPS-induced ALI mode and in LPS-induced RAW264.7 cells. Through molecular dynamics simulation, we observed that Pa was bound to the catalytic pocket of Akt and effectively inhibited the biological activity of Akt. These results indicated that Pa significantly relieves LPS-induced ALI by activating the Akt/NF-κB signaling pathway.

To investigate the molecular mechanism of modified Yigong powder(MYG)in the treat-ment of spleen deficiency syndrome based on network pharmacology and analyzed the effect of MYG on gastrointestinal simulated intestinal flora of spleen deficiency dogs and mucosal barrier of Caco-2 cells,as well as the interaction between intestinal flora and mucosal barrier.The molecular mechanism of MYG in the treatment of spleen deficiency syndrome was predicted by network pharmacology.The fecal samples of three canines(12±1)years old with spleen deficiency were collected to establish an in vitro gastrointestinal simulation system,which was divided into the o-riginal fecal sample group,the gastrointestinal simulation group and the gastrointestinal simulation treated by MYG group.The structural changes of the flora in each group were detected by 16S rD-NA sequencing.The metabolites were extracted from the gastrointestinal simulation system trea-ted by MYG group to study its effect on LPS-induced Caco-2 cell mucosal barrier injury model.The cell experiments included the blank control group,LPS model group,modified Yigong metabolite group.The permeability of mucosal was determined by fluorescein sodium,and then relative ex-pression levels of Claudin-1,Occludin and ZO-1 mRNA were determined by qPCR.The correlation between intestinal flora and Caco-2 cell mucosal barrier index after MYG intervention was further analyzed.The results showed that MYG had 76 active ingredients and 45 potential targets for the treatment of spleen deficiency syndrome.Forty key targets were obtained through protein interac-tion analysis,34 items were obtained by GO enrichment analysis,and 16 pathways were obtained by KEGG enrichment analysis.In the gastrointestinal simulation system,compared with the gas-trointestinal simulation group,at the phylum level,the abundance of Firmicutes,Bacteroides and Actinobacteriota increased significantly(P＜0.05),and the abundance of Proteobacteria decreased significantly(P＜0.05).At the genus level,the abundance of Fusobacterium,[Ruminococcus]gna-vus group and Blautia increased significantly(P＜0.05),while the abundance of Escherichia-Shi-gella and Citrobacter decreased significantly(P＜0.05).The diversity index of intestinal flora in the gastrointestinal simulation treated by MYG group was significantly increased(P＜0.05).In cell experiments,compared with the LPS model group,the mucosal permeability of Caco-2 cells in the modified Yigong metabolite group was significantly reduced(P＜0.01),and the expression levels of Claudin-1,Occludin and ZO-1 mRNA were significantly increased(P＜0.01).Correlation analysis showed that there was a certain correlation between bacterial community structure and mucosal barrier indexes.In summary,MYG may act on 40 key targets such as TNF,IL6,IL18,CX-CL8 and AKT1 through 76 active ingredients such as quercetin,arachidonic acid and naringin,and treat spleen deficiency syndrome in dogs through 16 signaling pathways such as AGE-RAGE,FoxO and HIF-l.In addition,the gastrointestinal metabolites of MYG up-regulate tight junction protein mRNA expression,reduce mucosal permeability,and repair mucosal barrier,which may be related to MYG's regulation of flora structure.

The negative effects of low temperature can readily induce a variety of diseases. We sought to understand the reasons why cold stress induces disease by studying the mechanisms of fine-tuning in macrophages following cold exposure. We found that cold stress triggers increased macrophage activation accompanied by metabolic reprogramming of aerobic glycolysis. The discovery, by genome-wide RNA sequencing, of defective mitochondria in mice macrophages following cold exposure indicated that mitochondrial defects may contribute to this process. In addition, changes in metabolism drive the differentiation of macrophages by affecting histone modifications. Finally, we showed that histone acetylation and lactylation are modulators of macrophage differentiation following cold exposure. Collectively, metabolism-related epigenetic modifications are essential for the differentiation of macrophages in cold-stressed mice, and the regulation of metabolism may be crucial for alleviating the harm induced by cold stress.
Acetylation ; Animals ; Cold-Shock Response ; Epigenesis, Genetic ; Macrophages/metabolism* ; Mice ; Mitochondria/metabolism*