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Influence of roflumilast on sepsis mice through the JAK/STAT signaling pathway.

OBJECTIVE: The aim of this study was to explore the influence of roflumilast on sepsis mice through the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway.

MATERIALS AND METHODS: A total of 36 Sprague-Dawley mice were randomly divided into normal group (n=12), model group (n=12) and roflumilast group (n=12). Mice in the normal group were fed normally. However, mice in the model group and roflumilast group were intraperitoneally injected with endotoxin to establish the sepsis mouse model. Furthermore, mice in the model group and roflumilast group were intraperitoneally injected with 0.9% sodium chloride and roflumilast once a day, respectively. After 7 d of intervention, mice were sampled. Lung tissue morphology was observed via hematoxylin-eosin (HE) staining, and the pathological score was given. The protein expression levels of JAK and STAT-3 were detected via Western blotting. The expression levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected via enzyme-linked immunosorbent assay (ELISA). Meanwhile, the mRNA expression levels of JAK, STAT-3, IL-6 and TNF-α were detected via quantitative Polymerase Chain Reaction (qPCR). The number of inflammatory cells in the lavage fluid was counted by a biochemical detector.

RESULTS: The survival rate of mice in the roflumilast group was significantly higher than that of the model group (p<0.05). The results of HE staining revealed that lung tissue morphology in roflumilast group was significantly improved when compared with the model group. Meanwhile, the pathological score in the roflumilast group was significantly lower than that of the model group (p<0.05). Western blotting showed that the protein expression levels of JAK and STAT-3 in the roflumilast group were markedly lower than those of the model group (p<0.05). According to the results of ELISA, the expression levels of IL-6 and TNF-α in the roflumilast group were remarkably lower than the model group (p<0.05). Further qPCR results manifested that the mRNA expression levels of JAK, STAT-3, IL-6 and TNF-α in the roflumilast group were significantly lower than those of the model group (p<0.05). Moreover, the number of neutrophils, monocytes and lymphocytes in the roflumilast group was significantly smaller than the model group.

CONCLUSIONS: Roflumilast can improve lung tissue morphology of sepsis mice by inhibiting the JAK/STAT signaling pathway.

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