The Study On Fingolimod (FTY720) Modulating Immunological And Inflammatory Functions To Improve The Prognosis Of Subarachnoid Hemorrhage(SAH)

ObjectiveSpontaneous subarachnoid hemorrhage(SAH)has a high morbidity and mortality.At present,there is still a lack of effective treatment for neurological dysfunction of SAH.To observe the changes of immune inflammatory reaction in early stage of SAH,and the application of new immunosuppressive agents Fingolimod intervention,explore new ways of treatment of SAH.MethodsPart One:In gene expression database(GEO)Download mouse cerebral basilar artery(GSE46696),using gene chip microarray analysis in sham operation group(Sham group)and model group(group SAH)samples of 3 copies of the gene variation,analysis and application of Cytoscape software on the differential expression of gene functional analysis and pathway enrichment analysis and gene-gene interaction.Part Two:The sham operation group(Sham group)and model group(SAH group)were 12,respectively,after modeling 1D,3D selected 6 mice brain tissue was detected by flow cytometry,natural killer cells(NK),CD4+ and CD25+ of endogenous regulatory T cells(Treg),respectively after modeling 1D and 3D are selected to detect brain tissues of 6 mice with Western Blot immune inflammatory factor,according to the analysis results of the first part of the thesis,3 significant changes in the relative immune inflammatory factor IL-1,TNF-?,NF-?B and TLR4 were detected.Part Three:The rats were divided into sham operation group(group Sham),model plus saline placebo group(group SAH+Saline),model plus fingolimod treatment group(SAH+Fingolimod group),SAH+Fingolimod group on the first day after modeling Fingolimod(FTY720)by intraperitoneal injection,the dose was 0.1 mg/kg,diluted to 1ml normal saline,dosing frequency was three times in 3 day,at 2h,48 h,72h after SAH.In 3 group model after administration of 1D,3D selected 6 mice,NK cells by flow cytometry in brain tissue model(NK),CD4+ and CD25+ of endogenous regulatory T cells;respectively in the 3 groups after administration of the 3D model from 6 mice with Western Blot the detection model of brain tissue inflammatory cytokines IL-1,NK-Beta Kappa B and TLR4 respectively in the 3 groups;model after administration of 1D,3D,7d,14 d selected 6 mice,detection of brain water content;in 3 group model after administration of 1D,3D,7d,14 d selected 6 mice were injected into the tail vein 2% Evans blue(EB)solution,detection of EB content in brain tissue;in 3 group model after administration of 1D,3D,7d,10 d,14d selected 6 mice with m NNS score and Morris water maze test,evaluate the neural function of mice.ResultsPart One:Analysis of gene chip and immune inflammatory factor SAH in mice model of cerebral basilar artery function,can be found in the immune inflammatory reaction at early stage of SAH may be important causes of poor prognosis,which significantly changes of IL-1 beta,TNF-?,NF-?B and TLR4 and other inflammatory factors may be SAH early brain injury(EBI)of a sensitive marker of severity.Part Two:SAH after the model of early NK cell percentage of CD3+ / T cells was significantly increased compared with Sham group,the difference was statistically significant(P < 0.05);group SAH CD4+ and Foxp3+ T cell and CD4+ T cell count ratio,compared with the Sham group increased slightly,there was no statistically significant difference(P > 0.05 Western;Blot)of brain tissue was detected by immune inflammatory factor IL-1 beta,TNF-?,NF-?B and TLR4 and Beta Kappa-action protein proportion was significantly increased,compared with Sham group,the difference was statistically significant(P < 0.05).Part Three:In early SAH,SAH+Saline group and SAH+Fingolimod group of NK cell percentage of CD3+ / T cells than in the Sham group,SAH+Fingolimod group were lower than SAH+Saline group,the difference was statistically significant(P < 0.05);group SAH+Fingolimod CD4+ and Foxp3+ T cell and CD4+ T cell counts than were higher than that in SAH+Saline group,the difference was statistically significant(all P < 0.05);SAH+Fingolimod group the expression of proinflammatory cytokines IL-1,NF-Beta Kappa B and TLR4 were lower than those in group SAH,group SAH+Fingolimod,inflammatory factors and-action protein were lower than those in group SAH,the difference was statistically significant(P < 0.05);the detection of 3D,water content in SAH+Fingolimod group 7d was significantly lower than that of SAH+Saline group,the difference was statistically significant(P < 0.05);the detection of 3D,EB in brain tissue of SAH+Fingolimod group was significantly lower than that of SAH+ 7d In Saline group,the difference was statistically significant(P < 0.05);SAH+Fingolimod group mNSS score was significantly lower in group SAH+Saline from the 3D,the difference was statistically significant(P < 0.05);application of Morris water maze,SAH+Fingolimod group cognitive function were significantly better than the SAH+ Saline group,acquired training and training on index the difference was statistically significant(P < 0.05).ConclusionsThe changes of immune cells and inflammatory cytokines play an important role in the brain injury and repair of SAH.EBI is a critical time for treatment of SAH brain injury.Fingolimod is a new immunosuppressant,can regulate immune function effectively,reduce inflammation injury,help to improve the prognosis of SAH,which provides a new way for the future of SAH regulation of early drug treatment,the implementation of good experimental basis for POC clinical research.