The Effect And Mechanism Of 3-(6-Phenylhexanoyl)Oxazolidin-2-One In The Central Inflammation

Background:For the past few years,as an anti-inflammatory and analgesic target,the role of N-acylethanolamine hydrolyzing acid amidase(NAAA)in the process of inflammation,pain,cancer and other diseases has been exploration and verification far more before.Inhibiting the activity of NAAA can enhance comcentration of PEA and OEA in tissues,that belong to fatty acid ethanolamine(FAE),thus existing a certain degree of anti-inflammatory,analgesic,antiepileptic and neuroprotection.However,Most of the existing NAAA inhibitors are local application,show the low activity and the short half-life,which not conducive to oral administration and whole animal experiments,discourage the use of NAAA inhibitor.So it is essential and of grate value to find potent NAAA inhibitor.Objective:Our lab has synthesized a potent stable and highly selective NAAA inhibitor named F96 with low toxicity,In this study,we have established a series of experiments to determine the anti-inflammatory effect of F96 in the brain,trying to find out the way how the drug works,and explore the mechanism of effect.Method:Based on the synthesized of F96 in our group,we use LPS caused animal model of short-term acute brain inflammation and long-term chronic brain inflammation.The content of major inflammatory factors in serum and brain was determined by ELISA.Use real-time PCR to detecte the expression of inflammatory factors and NAAA,NAPE-PLD,PPAR-a on genetic in tissues.Use Western blot to analyze the protein expression of the inflammatory factors,and marker molecule Iba-1 activated by microglia.Using the LC-MS technique to amount the change of fatty acid ethanolamine(FAE).The brain injury and cell migration were observed by hematoxylin-eosin staining and the permeability of blood-brain barrier was measured by Evans blue.The activation of microglia and astrocytes were observed by immunofluorescence.The changes in intestinal flora were measured by Genome sequencing,which cause some of effect on brain..BV-2 cells were cultured to determine the effect of the drug.Results:F96 is useful to decrease the expression of TNF-a in serum in the model of administration LPS for 1 hour.F96 can reduce the expression of inflammatory factors in brain on genetic in c57/B6 mice.F96 can restore the content of PEA,partially elevated OEA content in the brain of inflammatory mice,but does not affect other endogenous lipids.F96 can inhibit the activation of microglia and astrocytes.F96 can rectify the alteration of intestinal flora caused by LPS.Conclusions:F96 has a certain therapeutic effect on acute and chronic brain inflammation caused by LPS.It's can be achieved by inhibiting the activation of microglia and astrocytes and inhibiting the transfer of inflammatory factors from plasma to brain.