| BACKGROUND AND PURPOSEBasic fibroblastic growth factor (BFGF) is an effective neurotro-phic and vasoactive polypeptide. The objective of this research is to investigate BFGF expression and cellular dislocation at different time after transient induction of focal cerebral ischemia, evaluate effect of BFGF on NO and explore its protective effect to ischemic brain tissue.METHODS1. source of animals and grouping : 55 healthy male wistar great gerbil, weighing 280 30g were randomly divided into control group and experiment group. (1) control group; normal control group( n = 5). (2) experiment group: (1) ischemia group was divided into 6 groups; group 1, 6h of reperfusion after 120 min of ischemia; group2, 12 h of reperfusion; group 3, 24 h of reperfusion; group4, 3 d of reperfusion; group 5, 4 d of reperfusion; group 6, 5 d of reperfusion (n =5 in every group). (1) ischemia treatment group (intraperitoneal injection of 100 gKg-1 BFGF dissolved in saline immediately after onset of ischemia) was divided into 4 groups: group 1: 6h of reperfusion ; group 2:12 h of reperfusion; group 3 ;24 h of reperfusion; group4: 72 h of repeifusion ( n =5 in every group) , ischemia group as control group (intraperitoneal injection of saline).2. MCAO model I use a transient middle cerebral artery { MAC) occlusion model. The right carotid region was exposed through a cervical midline skin incision and division of the right omohyoid muscle. The external carotid artery ( EGA) was coagulated and divided after trimming the proximal EGA branches. The internal carotid artery (1CA) was then exposed and the pterygopalatine artery was ligated with asilk suture. After clamping the common and internal arteries,a small incision was made in the EGA and a monofilament suture with a rounded tip was introduced and advanced approximately 18 0. 5mm into the IC A through the carotid bifurcation for proximal MCAO.3. Immunochemistry Temporal and cellular changes of immuno-histochemical BFGF expression were compared with different priods of repeifusion from 6 hours to 7 days after transient MCA occlusion. Coronal section were cut with 6 umthicknesss and mounted on standard glass slides. Deparaffinvation was a chieved by treating the specimens in xylene and subsequent ethano followed by a rinse in PBS. After they were blocked with 10% normal horse serum for 2 hours. The slides were washed in PBS and incubated with a rabbit polyclonal antibody against BFGF at 1:50 dilution for L6 hours at 4 subsequently they were incubated with avidinbiotinnorsemdish peroxidase complex for 30 minutes and then developed with the use of DAB as a color substrate. The reaction was stopped by washing the slides in distillled water. To ascertain specific binding of the andibody for the protein a set of brain sections was stained in similar way with 0.01MPBS instead of the first antibody.4. NO determination: heart blood of all samples were taken be-fore reperfiision, 2.0 ml of 75 mmol/L ZnSO4 was added. Blood plasma was separated after shaking and 10 min of centrifugation, divided in different tubes and preserved under - 20 . Clock method was a-dopted.RESULT1 Immunochemistry result(1) BFGF immune positive cell was found in negative control group.(2) BFGF immune positive cell expressed in neuron and ganglial cell.(3 ) Compared with control group, expression of BFGF immune positive cell increased obviously at 6 h, 12 h, 24 h, 1 d, 3 d, 4 d, whether infarction region or not , neuron or ganglia! cell. The expression got to climax at 1 th d, decreased from 3 th d, and had only a less expression at 7 th d without expression at ischemic contralateral side.(4) Expression of BFGF immune positive cell.(5 ) BFGF immune positive cell was found in cerebral pia mater ependyma and choroid plexus.2 NO determination resultCompared with control group, NO levels in drug - taking group at 12 h, 24 h, 3 d of reperfusion decreased remarkably. After drug -taking, NO level increased slowly with the developing of ischemic reperfusion...
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