| Objective To investigate the effect of different doses of DiZhi Beef Decoction(DZBD)in peripheral splenic T-cell subpopulation and NK cell of rats with cerebral ischemia,and understand the potential mechanism of barin protective throng the targeted control of the immune function of spleen,providing theoretical basis for clinical treatment and the theoretical guidline of diateryin ischemic brain patients.Methods Eighty male SD rats were randomly divided into sham group(n=20),vehicle group(n=15),low-dose DZBD-treated group(n=15,200mg·kg-1·d-1),medium-dose DZBD-treated group(n=15,400mg·kg-1·d-1)and high-dose DZBD-treated group(n=15,600mg·kg-1·d-1).Sham group was divided into two subgroup:sham group I(n=10)and sham group Ⅱ(n=10)..Through the intraluminal filament method,permanent focal cerebral ischemia model was induced in vehicle group and DZBD-treated group.For 7 consecutive days,the rats were lavaged different doses of DZBD in DZBD-treated group,and the other rats were received saline water in sham group and vehicle group.The body weight was observed before surgery and 1 d、3 d、7 d.The neurological function was evaluated with longa 5-point scale in 2 h、1 d、3 d and 7 d.After 7 d,the spleen mass was weighted and the spleen index was calculated.The volumes of infarction were observed by TTC and brain histopathology was evaluated by HE staining.The percentage of T lymphocyte(CD3+、CD4+and CD8+)and NK cell in spleen were measured by the method of flow cytometry and the ratio of CD4+/CD8+was calculated.Results1.Compared with sham group,the body weight was significantly decrease in 1 d、3 d、7 d(P<0.01).After lavaged high-dose DZBD treatment,it was up-regulated(P<0.05)in 3 d and 7d.2.Neurological dysfunction scores were no difference in 2 h(P>0.05).Compared with vehicle group,neurological dysfunction scores were decreased with different dose of DZBD-treatment in 3 d and 7 d(P<0.01,P<0.05).3.After 7 days,compared with vehicle group,infarct volumes were decreased in different doses of DZBD-treated groups,especially in medium-dose DZBD-treated group and high-dose DZBD-treated group(P<0.01).4.HE staining showed that neurons pathological injury of cerebral cortex was obvious in vehicle group.But it was lighter in different doses of DZBD-treated groups.5.Compare with sham group,the spleen mass and the index of spleen were reduced in 7 d(P<0.01).With different doses of DZBD-treatment,the index of spleen was up-regulated in different doses of DZBD-treated groups(P<0.01),meanwhile,the spleen mass was increased in medium-dose and high-dose DZBD-treated group(P<0.05,P<0.01).6.Compared with sham group,the percentage of CD3+and CD4+,the ratio of CD4+/CD8+were all decreased(P<0.01,P<0.05,P<0.01),but there was no difference in CD8+(P>0.05).Compared with vehicle group,the percentage of CD3+(P<0.01)and CD4+/CD8+(P<0.01,P<0.01,P<0.05)also could be up-regulated in DZBD-treated groups.7.After 7 d,the level of NK cell was decreased in vehicle group(P<0.01).With different doses of DZBD,the level of NK cell was up-regulated in medium-dose DZBD-treated group and high-dose DZBD-treated group(P<0.01,P<0.05).8.The infarct volume had a negative correlation with the pencentage of T lymphocyte(CD3+、CD4+)(r=-0.655,P=0.002;r=-0.451,P=0.046)、CD4+/CD8+(r=-0.546,P=0.013)and NK cell(r=-0.512,P=0.021),but there was no relative between infarct volume with CD8+(r=0.292,P=0.212).Conclusion1.DZBD has a neuroprtective effect through sustain body weight,improve neurological dysfunction symptom,reduce infarct volumes and relieve pathological injury in ischemia brain.2.The spleen mass and the index of spleen can be up-regulated with DZBD,meanwhile,the percentage of T lymphocytes and NK cells are increased in spleen.Therefore,DZBD can make a protective role in peripheral immunity dysfunction after cerebral ischemia of rats.3.The infarct volume has a negative correlation with T lymphocyte and NK cell on spleen,the peripheray immunosuppression will make a damage to iachemia brain. |
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