The Effect Of Tubastatin A On The Interaction Of HSP90 And INOS In LPS-stimulated RAW264.7

Objective: The effect of histone deacetylase 6 inhibitor tubastatin A on the interaction between the level of acetylation of heat shock protein 90 and inducible nitric oxide synthase activity by lipopolysaccharide-stimulated macrophage.Methods: Macrophage RAW264.7 cells in mouse can be divided into control group,the LPS stimulation group,LPS + tubastatin A treatment group.In 6 hour,8 hours and 10 hours collect cells and cell culture.Western blots were performed to acess protein levels of HSP90,acetylated HSP90 and iNOS.Griess assays were utilized to analyze the release of nitric oxide(NO)respectively in the cell culture medium.Results: In the experimental observation,the level of LPS stimulus RAW264.7 cell HSP90 expression,there is no obvious change,but the level of acetylation of HSP90 intensity increased.The level of iNOS expression of LPS group in 6 hours,8 hours and10 hours significantly increased than that of control group,and the level of NO in cell culture medium were in the same trend.In the experimental observation,there were no dramatic change of HSP90 level of expression when the LPS group compared with LPS+Tubastatin A group.But the acetylation of HSP90 significantly higher in LPS+Tubastatin A group in comparison to LPS group.There were no clear differences of level of iNOS expression between LPS group with LPS+ Tubastatin A group.But the level of NO in cell culture,LPS group significantly lower than LPS+ Tubastatin A group.Conclusion: LPS decreased the acetylation level of HSP90 in macrophages,increased the activity of HSP90,and increased the protective effect of HSP90 on the activity of iNOS,thus stabilizing the activity of iNOS.Tubastatin A restored the acetylated level of HSP90 reduced by LPS and reduced the activity of HSP90,resulting in a weakened protective effect on the activity of iNOS,thereby inhibiting the activity of iNOS.