The Effects And Mechanisms Of Compound LX007 On Lipopolysaccharide-induced Inflammation In Primary Microglial Cells

Objective:It is known that over-activated microglial cells play an important role in various neurological diseases,such as ischemic stroke,Alzheimer's disease,Parkinson's disease and multiple sclerosis.In order to find better drugs to cure those diseases,a series of new compounds have been designed and synthesized in the Department of Medicinal Chemistry,School of Pharmacy,Nanjing Medical University,which has a hydrophobic ring A and a hydrophilic ring B with a carboxyl group.Previous reports have shown that ZL006,a synthesized compound,exhibited strong neuroprotective activity in vitro and in vivo.However,the anti-inflammatory effects of these compounds in the central nervous system have not been elucidated.4-((5-bromo-3-chloro-2-hydroxybenzyl)amino)-2-hydroxybenzoic acid(LX007),one of the members of these newly synthesized compounds,was used to evaluate the anti-inflammatory effects in lipopolysaccharide(LPS)-induced inflammatory responses of primary microglia.Methods:The present study investigated whether LX007 could exert an anti-inflammatory effect on LPS-induced mouse primary microglia cells.Real-time PCR(rt-PCR),western blotting(WB)and enzyme linked immunosorbent assay(ELISA)were used to detect the expression of pro-inflammatory mediators.In addition,the potential underlying mechanisms of LX007 on inflammation were examined,which might involve the mitogen-activated protein kinase(MAPK)and nuclear factor-kappa B(NF-?B)signaling pathways.Results:The main results of this research are as follows:1)LX007 could decrease the expression of inducible NO syntheses(iNOS)and cyclooxygenase-2(COX-2)in LPS-stimulated primary microglia;2)LX007 could reduce the production of nitric oxide(NO)and prostaglandin E2(PGE2)in primary microglia treated by LPS;3)LPS-induced over-production of pro-inflammatory cytokines including interleukin-1?(IL-1?),IL-6 and tumor necrosis factor-?(TNF-?)from microglia were also significantly attenuated by LX007;4)Mechanistically,it was found that LX007 could down-regulate the activation level of MAPK and NF-?B pathway of LPS-induced inflammation in primary microglia.Conclusion:LX007 exhibits anti-inflammatory effects in LPS-stimulated primary microglial cells by reducing pro-inflammatory mediators,which might be associated with down-regulating of the activation level of MAPK and NF-?B pathway.