Department of Infectious Diseases, the First Affiliated Hospital of Anhui Medical University, Hefei, China.
Eur Rev Med Pharmacol Sci. 2019 Feb;23(3):1335-1341. doi: 10.26355/eurrev_201902_17028.
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.
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.
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.
Roflumilast can improve lung tissue morphology of sepsis mice by inhibiting the JAK/STAT signaling pathway.
本研究旨在通过 Janus 激酶/信号转导和转录激活因子（JAK/STAT）信号通路探讨罗氟司特对脓毒症小鼠的影响。
将 36 只 Sprague-Dawley 小鼠随机分为正常组（n=12）、模型组（n=12）和罗氟司特组（n=12）。正常组小鼠正常喂养，模型组和罗氟司特组小鼠均腹腔注射内毒素建立脓毒症小鼠模型，同时模型组和罗氟司特组小鼠分别每日腹腔注射 0.9%氯化钠溶液和罗氟司特。干预 7 d 后取材，苏木精-伊红（HE）染色观察肺组织形态，给予病理评分；Western blot 检测 JAK 和 STAT-3 蛋白表达水平；酶联免疫吸附试验（ELISA）检测白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）表达水平；实时荧光定量聚合酶链式反应（qPCR）检测 JAK、STAT-3、IL-6 和 TNF-αmRNA 表达水平；生化检测仪检测肺泡灌洗液中炎症细胞数量。
罗氟司特组小鼠存活率明显高于模型组（p<0.05）。HE 染色结果显示，罗氟司特组小鼠肺组织形态较模型组明显改善，病理评分明显低于模型组（p<0.05）。Western blot 结果显示，罗氟司特组 JAK 和 STAT-3 蛋白表达水平明显低于模型组（p<0.05）。ELISA 结果显示，罗氟司特组 IL-6 和 TNF-α表达水平明显低于模型组（p<0.05）。qPCR 结果显示，罗氟司特组 JAK、STAT-3、IL-6 和 TNF-αmRNA 表达水平明显低于模型组（p<0.05）。此外，罗氟司特组中性粒细胞、单核细胞和淋巴细胞数量明显少于模型组。
罗氟司特通过抑制 JAK/STAT 信号通路改善脓毒症小鼠肺组织形态。
