| Objective:Based on Xuanfu theory,to study the effects of Opening Xuanfu method on neurologic impairment,cerebral tissue water content,superoxide dismutaseactivity?SOD?,malondialdehydecontent?MDA?,matrix metalloproteinase-9?MMP-9?,hemorheology,permeability and ultrastructure of blood-brain barrier?BBB?after intracerebral hemorrhage in rats.And it will reveal the mechanism of opening Xuanfu method on regulating brain edema and BBB,then to provide the basis for the theory of"Xuanfu-Blood-brain barrier".Methods:1.Groups:180 male Sprague-Dawley rats were randomly divided into sham-operation group,model group,low,medium and high dose groups,with 36rats in each group.2.Dosing:low,medium and high dose groups were 2.77g/kg·d-1,5.53g/kg·d-1,11.06g/kg·d-1.3.Modeling:the model of intracerebral hemorrhage was established by injecting 50ul of autologous caudate artery into caudate putamen nucleus,using rat brain stereoscopic locator.4.Sampling and testing:At 6 hours,3days and 7 days after operation,Blood samples were collected from abdominal aorta to measure hemorheology;Brain water content was measured by dry and wet weight method;SOD activity and MDA content were measured;The brain tissue was taken to make paraffin sections after cardiac perfusion,HE staining;The expression of MMP-9 was detected by immunohistochemistry;Intravenous injection of Evans Blue through the tail,perfusion fixation,The permeability of blood-brain barrier was detected by taking brain tissue,ultrastructure of blood-brain barrier was observed by electron microscope.Results:1.Neurological impairment score:Compared with the sham-operation group,the score in the model group was significantly higher than that in the sham-operation group at each time points?P<0.05?.Compared with the model group's score:the low dose group decreased at each time points?P>0.05?;the medium,high dose group decreased at each time points,but the high dose group significantly decreased at all time points?P<0.05?.2.The brain water content?BWC?:Compared with the sham-operation group,the model group's BWC increased and reached the peak at 3d?P<0.05?,and still higher than sham-operation group at 7d.Compared with the model group:the low dose group's BWC decreased at 3d and 7d,but the difference was not significant?P>0.05?;the medium dose group's BWC was lower than model group at 3d?P<0.05?,and the high dose group's BWC was significantly lower than that of the control group at 3d after 7d?P<0.05?.Compared with the low and medium dose groups,the high dose group's BWC was significantly decreased at 3d and 7d?P<0.05?.3.EB content:?1?In bleeding side:Compared with sham-operation group,the EB content in the model group increased at all time points?P<0.05?,and reached the peak at 3d?P<0.05?.Compared with model group's EB:the low dose group decreased at 6h and 3d?P<0.05?,and slightly decreased at 7d?P>0.05?;the medium and high dose group decreased at each time point?P<0.05?,the decrease was most obvious in the high dose group?P<0.05?.?2?In non-bleeding side:Compared with sham-operation group,the model group's EB increased at each time points?P<0.05?,and reached the peak at 3d?P<0.05?.Compared with the model group's EB,the low and medium dose group slightly decreased at 6h?P>0.05?,increased at 3d?P<0.05?but decreased at 7d?P<0.05?;the high dose group increased at 6h and 3d?P<0.05?,but decreased at 7d?P<0.05?.4.SOD activity in brain tissue:Compared with the sham-operation group,the model group's SOD activity was lower than sham-operation group at each time points?P<0.05?,and decreased to the lowest at3d?P<0.05?.Compared with the model group,the SOD in the low and medium dose group was higher than model group at each time point?P<0.05?,and high dose group's SOD activity was higher than model group at 6h and 3d,but significantly decrease in 7d?P<0.05?.5.MDA content in brain tissue:Compared with the sham-operation group,the model group increased significantly at each time points?P<0.05?,and increased to the highest at 3d?P<0.05?.Compared with model group's MDA:the low and medium dose groups decreased at each time point?P<0.05?;the high dose decreased at 6h and 3d?P<0.05?,but it showed an increasing trend at 7d?P>0.05?.6.Hemorheology:Compared with the sham-operated group,the whole blood viscosity?lower,and the medium and high shear rates?and red cell assembling index of the model group were higher?P<0.05?.Compared with the model group,the whole blood viscosity and red cell assembling index of each administration groups were decreased?P<0.05?,and he high dose group were significantly decreased?P<0.05?.7.The expression of MMP-9:Compared with sham-operation group,there was no significant expression in model group at 6h,but increased at 3d and 7d?P<0.05?,and reached the peak at 3d?P<0.05?.Compared with the model group,each dose group's expression of MMP-9increased at 6h,but decreased at 3d and 7d?P<0.05?,and the change was obvious in the high dose group.8.Ultrastructure of blood-brain barrier around hematoma:Model of the basement membrane was irregular shape,uneven thickness,endothelial cell swelling,mitochondrial swelling,cristae.Compared with the model group,the structural abnormalities of basement membrane,endothelial cells and mitochondria were improved in the low,medium and high dose groups,and the edema around the blood vessels was alleviated,especially in the high dose group.Conclusion:1.Jiajian Qufeng Tongqiao fang can alleviate the nerve function defect,reduce the water content of brain tissue,and improved the damage of the BBB structure on the bleeding side after cerebral hemorrhage in rats.2.The main mechanism may be to improve microcirculation,enhance the activity of SOD,reduce the content of MDA,reduce the expression of MMP-9,so as to reduce the damage of blood-brain barrier,reduce the permeability of blood-brain barrier,and reduce the brain edema.3.The high dose of Qufeng Tongqiao fang can decrease the SOD activity and increase the content of MDA in brain tissue,which may be related to the toxic effect of ephedra.Therefore,this prescription should avoid high dose use or long-term use.4.The permeability of blood-brain barrier was increased in the hemorrhage side and non-bleeding side after intracerebral hemorrhage.The method of opening Xuanfu had a bidirectional effect on the blood-brain barrier after cerebral hemorrhage.That is,the permeability of blood-brain barrier in the bleeding side was decreased.The blood-brain barrier permeability of the non-bleeding side was increased at a certain stage,but whether the opening effect of the drug can promote the drug into brain tissue to enhance the curative effect still needs to be further studied. |
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