| AIMBlood Brain Barrier ( BBB) is constituted with the tight junction of endo-thelial cell, basal lamina and astrocyte feet . It is a interface located in the brain and the capillaries to regulate the materials that enter in the brain tissue. The destruction of blood - brain barrier(BBB) is a important pathophysiology mechanism of cerebral ischemia - reperfusion injury. At the earlier period of the cerebral ischemia reperfusion, large quantity of cytokine and adhesive molecules, such as IL - 1, TNF, IL -6, IL -8,etc is expressed, and transform the is-chemic injury of brain into inflammation reaction. While IL - 10 has the function of protection. Inflammation and immune response have an important role in pathophysiology mechanism of cerebral ischemia-reperfusion injury, involving the infiltration of leucocyte and the activation of macrophage. The activation of leucocyte can produce much proteolytic enzyme, oxygen - derived free radidicals and other factors, cause the destruction of capillary endothelium and basal mem-bran , and increase the permeability of blood - brain barrier.Hyperbaric oxygen ( HBO) , as a kind of valid method of no wound , is applied quite broadly in the treatment of brain disease. As it can ameliorate the situation of hypoxia , Maintein the energy metabolism and relieve encephaledema. While The study of the effect of HBO on blood - brain barrier after cerebral ischemia ?reperfusion has been rare. This study in cellular, molecular and gene level respectively explored the effect of HBO on the permeability of blood - brain barrier after cerebral ischemia - reperfusion and found the evidence which HBO can therapy cerebral vessel disease.MethodsThe replication of cerebral ischemia - reperfusion animal models: conscious mouse , performing surgical operation on neck to separate the double common carotid artery and fix with plasticine. Then tensing the silk thread to make theblood flow of artery be blocked, 30minutes later,loosing the line and suturing the skin sterily. The HBO treatment of experimental animals: HBO group and HBO + cerebral ischemia - reperfusion group entered the HBO cabin after reperfusion 2 h,9 h,21 h,45 h,69 h, then washing the cabin 10 minutes with pure oxygen to make the concentration of O2more than 90% , in the velocity of 0.0125 MPa/ min, maintaining the pressure to 0.25 Mpa for 60 minutes.The cerebral blood flow measurements: Opening the cranial cavity, and use laser doppler blood - flow measuring equipment to monitor the normal brain tissue and the ischemia 10 minutes,20 minutes,30minutes ,ischemia reperfusion 4h,11h,21h,48h,72h brain cortex blood flow.The content of Evans blue in brain tissue; As the Method of Udaka K. etc, Before 1 hour of death, the mouse was injected through tail vein 2% Evans blue , 20 minutes before examination, using saline to filling the heart before the atrium run off clear liquid. Putting the mouse to death at reperfusion 4h,11h,23h, 48h,72h and harvesting the brain , using the electronics weighing scoles to weight and put them into tube. 3 ml formamide was adding into them. Tube was covered and put in 45 C water bath for 48 h, lightly shake them and centrifuga-tion for 15 min(3000 rs/ min) , take the pure liquid at 722 Spectrophotometer. ( X= 632 nm).Using ABC - ELISA method to measure the content of cytokine; 72 hours after surgery, the mice was put to death and quickly took out of the brain on a ice dish, washing them with ice PBS and suck dry with filter paper, immediately u-sing the electronics weighing scoles to weight and make 10% tissue homogenizer , 4 C 4000 rs/ mins centrifugation for 15 minutes, and taking the pure liquid, using ABC - ELISA in the wave - length of 492 nm to examine the IL -1, TNF, IL-6, IL-8 andIL - 10.Gelatin zymography to detect the activity of gelatinase : Taking the hippocampus of brain tissue and making 10% tissue homogenizer, then centrifugation to obtain protein extraction. Taking out of 20ul and add in the gel(5mg/ml gelatin) ,20mA constant current electrophoresis for 5h,then letti...
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