Protective Effect Of CD4~+CD25~+Regulatory T Cells On Blood-brain Barrier And Apotosis By Transplanting Into Intracerebral Hemorrhage Mice

ObjectiveEstablishment a treatment model of Intracerebral hemorrhage mice,to rese-arch the immunomodulatory cerebroprotective effects of exogenous CD4+CD25+regulatory T cells（CD4+CD25+Tregs） after transplanting through femoral veinby observing the related indicators.MethodsIn this experiment,select10-12weeks old,weighing about25g,Kunming ad-ult male mice for the study.All of the animals were randomly divided into:ICHsurgery group and sham-operated (sham) group.In the surgical group,injectionof10ul autologous whole blood into the mice’ left striatum using the improvedmethod.Magnetic cell sorting method to extract CD4+CD25+Tregs from mice’spleen.Establishment a model of regulatory T cells treatment by transplantinginto the mice through femoral venous.And they were divided into three subgrou-ps:Tregs group,CD4+CD25+Tregs were transplanted into the mice;PBS group:e-qual volume of PBS were injected;ICH group:without any special treatment aft-er cerebral hemorrhage.Corner turn and foot fault tests were used to assess thebehavioral for one month.Blood-brain barrier permeability was determined byEvans blue leakage method.Enzyme-linked immunosorbent assay(ELISA) wasused to evaluate the expression level of the related indicators of free radicals ofMDA、 SOD in the brain tissue from one day to one week after ICH. labelingapoptosis-related indicators with Caspase-3and Tunel technology.Results 1.Compared with the sham group,The mice showed different degrees ofneurological deficits from the first day after cerebral hemorrhage,especially inthe ICH and PBS group(P<0.05),they need one month to restore.The corner turnand foot fault tests’ score of Tregs were significantly lower than others(P<0.05).they can be returned three weeks after ICH.2.The related indicators of apoptosis around the hematoma detected byImmunofluorescence microscopy showed that:Compared with sham group，in thesurgical group,the fluorescence intensity of TUNEL and Caspase-3were signifi-cantly higher(P<0.05),especially in the ICH and PBS group (P<0.001).Besides，the leakage of vascular EB,the expression level of free radicals of MDA at eachtime point on mice cerebral hemorrhage models were also increased significant-ly(P<0.05),and nothing changing after one week.But SOD was reverse lower(P<0.05);Tregs group,although have similar changes,but not as significant as othergroups(P<0.05).Conclutions1.using the improved autologous injection method can successfully constr-uct a model of cerebral hemorrhage mice;Tregs can be successfully transplantedinto mice through the femoral venous.2.Exogenous CD4+CD25+Tregs can significantly promote the recovery ofneurological function of ICH mice.3.Exogenous CD4+CD25+Tregs can significantly decrease the damage ofBBB after ICH.4.Exogenous CD4+CD25+Tregs can significantly reduce the level of MDA,the consumption of SOD,the inhibition of apoptosis,reduce damage,play a cereb-roprotective effect.