Fasudil Mediates NLRP3 Inflammasome To Protect Mice From Klebsiella Pneumoniae

Objectives:The aim of this study is to investigate the relationship between the well-studied inflammatory molecule complex of NLRP3 inflammasome and the RhoA/ROCK signaling pathway,which plays a versatile role in human body.We investigated that whether Fasudil,a clinical used ROCK inhibitor up to date,acts on Klebsiella pneumoniae-induced inflammasome activation and explored its mechanism.In this study we tried to explore the host's in-depth mechanism of immune response to bacterial infection,to find the factors potentially affecting the prognosis of infectious diseases which used to be unclear,and to enrich the theoretical basis for the pathogenesis and mechanism of infectious diseases in order to explore potential therapeutic targets finally,Methods:Through in vitro and in vivo experiments,we carried on this research exploring the truth.In vivo,we monitored a 5-day survival rates of mice suffering from pneumonia.We observed the levels of inflammatory cytokines and NLRP3 inflammasome activation in lung tissue,serum and BALF by H&E staining,Western Blot,ELISA and qPCR and probed into its possible mechanism whether Fasudil shows a prognostic function in mice with pneumonia.In vitro,bone marrow-derived macrophages(BMDM)from wild-type mice were used in the condition of Klebsiella pneumoniae invasion.By means of WB and qPCR we detected the inflammatory cytokines and inflammasome activation in both translational level and post-translational levels.Results:? Fasudil protects against pneumonia caused by Klebsiella pneumoniae in mice.? Fasudil attenuates.lung injury in mice with Klebsiella pneumoniae.? Fasudil decreases the expression of BALF IL-1? levels and reduce lung inflammation leakage.? Fasudil inhibits NLRP3 inflammasome activation in bone marrow-derived macrophage.? Fasudil alleviates the expression of inflammasome in mouse alveolar macrophage.Conclusion:Fasudil did protect mice in a reduced mortality and from lung injury induced by Klebsiella pneumoniae and reduced the levels of inflammatory cytokines.In agreement with the in vitro assay,the studies in vitro confirmed that Klebsiella pneumoniae indeed activates NLRP3 inflammasome in BMDM,whereas fasudil attenuates this activation in translational level and protein levels.