The Regulatory Effcet Of Macelignan On Microglia Mediated Neuroinflammatory Response

Objective:To investigate the effect of macelignan on the polarization phenotype of M1 microglia and the regulation of LPS induced microglia conditioned medium on neuronal activity.Methods:Lipopolysaccharide was used to induce over activation of microglia.The polarization phenotype of microglia and its regulation on neuronal activity were studied by intervention of macelignan in different concentrations.① MTT colorimetry was used to detect the effect of different concentrations of macelignan on the activity of microglia and neurons;② Griess method was used to detect the effect of no content in the supernatant of activated microglia;③ The expression of iNOS and FOX03a in microglia and CyclinDl and p27 in hippocampal neurons were observed by fluorescence staining;④ The expression of p-pi3k,p-Akt,p-foxo3a and p-mTOR in activated microglia was detected by Western blot.⑤The effects of macelignan on survival rate,morphology and expression of cell cycle related proteins in rat nippocampal neurons were examined.Results:1.The results of MTT showed that macelignan with final concentration of 2.5-20μm had no significant difference in the survival rate of microglia and neurons compared with the control group,indicating that it had no toxic effect on cells.2.Griess method showed that there was a significant difference between the control group and the BV2 cells treated with 1 μg/ml lipopolysaccharide(P<0.0001);with the increase of macelignan concentration,the NO concentration in the supernatant of microglia cells gradually decreased,compared with the lipopolysaccharide group,P<0.001 or P<0.05,indicating the success of the inflammatory cell model and the anti-inflammatory effect of natural drugs.Immunofluorescence staining and Western blotting also showed that macelignan could inhibit the expression of iNOS in LPS induced microglial BV2 cells,demonstrating the polarized phenotype of M1 microglial cells.4.Western blotting showed that macelignan inhibited the inflammatory response of microglia induced by lipopolysaccharide by regulating the phosphorylation of Akt,mTOR,FOXO3a and other proteins.5.Lps-mcm(lps-mcm)was used to treat HT22 cells.The results showed that macelignan inhibited the necrosis of HT22 cells induced by lps-mcm by regulating the expression of cyclin D1 and p27.Conclusion:1.Macelignan inhibits M1 microglial mediated neuroinflammatory response.2.Through PI3K/Akt pathway,macelignan may regulate the activation of mTOR,FOXO3a and other target proteins,inhibit neuroinflammatory response and play a protective role on neurons.3.Macelignan can inhibit the damage of neurons by endogenous neurotoxin.