Study on the Role of Epilepsy Activation Smad Signaling in Promoting M2 Type Polarization of Microglia
Objective To investigate the effect and mechanism of epilepsy on M2 type polarization of microglia in hippocampus of mouse brain tissue.Methods Forty C57 mice were randomly divided into control group(CON group,12 mice)and epilepsy group(PTZ group,28 mice).CON group was given normal saline,PTZ group was given pentylenetetrazole by intraperitoneal injection every other day for 28 days.After the experiment,the samples were collected and the corresponding experiment were carrried out.During the process of inducing epilepsy,the behavior,seizure level,duration and outcome of the mice were observed.HE staining and Nissl staining were used to observe the morphology and survival of cells in the hippocampus of mouse brain tissue.Immunohistochemical staining was used to observe the morphological changes of microglia in the hippocampus of mouse brain tissues.Western blot was used to detect the expression of Iba-1,Bax,Bad,p-Bad,Cleaved caspase-3,Arg-1,CD163,CD206,M-CSF,GFAP,Smad2,Smad3,p-Smad3,and Smad4 proteins in mouse brain tissue.Results HE staining and Nissl staining showed that the cells in CON group were round and arranged neatly.The cells in the hippocampus of PTZ group were deeply stained and constricted,with irregular shape and disordered arrangement.Western blot results showed that the protein expressions of Bax,Bad,p-Bad,Cleaved caspase-3,GFAP increased in brain tissue of PTZ group(Pall＜0.05).The protein expression of microglia marker Iba-1 increased,and the protein expression of M2 markers Arg-1,CD163 and M-CSF increased(P all＜0.05),but no significant difference was found in CD206.The protein expressions of Smad2,Smad3,p-Smad3 and Smad4 increased(P all＜0.05).Immunohistochemical results showed that the morphology of Iba-1 positive cells in the hippocampus of CON group was branched and clearly branched,while the morphology of Iba-1 positive cells in the hippocampus of PTZ group was branched,debranched or ameboid.Conclusion Epilepsy promotes apoptosis in hippocampus of mice,leads to brain injury,increases the expression level of Smads protein,and promotes microglial polarization to M2.
万方数据
维普
