The Expression Of BFGF With Bcl-2 Gene Injected After The MCAO For 2 Hours In Rat

ObjectiveTo investigate the expression of bFGF in rat with bcl - 2gene injected after the MCAO for 2 hours was induced and to study the effect of bcl - 2 against ischemia brain of the rats and if its role related to the expression of bFGF following cerebral ischemia in rat.Methods99 Wistar rats of 280 - 300g were randomly assigned on 3 groups: control group( control group n = 33 ) , bcl - 2 - treated group ( bcl - 2 group n = 33 ) , pLXSN group(plasmid control group n =33) ,each group was divided into randomly again 3 second group,24h,48h and72h after referfusion. First,temporary ischemia of the left middle cerebral artery occlusion( MCAO)for 2 hours was induced by the suture occlusion technique, took out the suture after 2 hours. The right side of the rats body movable obstacle, mean the model success. Second, we injected pLXSN plasmid and pLXSN - bcl -2 plasmid into the rats' left internal carotid artery soon after ischemic reperfusion 3 hours. Final,we measure these following things to the rats injected successfully and carried on the covariance a-nalysis. 1. The cerebral infarction volume: The rats who measure the infarction volume broke the head and took the brain directly, adopted the brain each 2mm slice,the infartion part presented white,the normal part assumed the red. We detect the cerebral infarction volume after 24,48 and 72 hours of cerebral ischemic reperfusion by TTC staining ,4% multitude gather formaldehyde and image anal-ysis system. 2. immunohistochemistry of bcl -2 and bFGF: 10% the water match chlorine alehyde,3. 5ml/kg belly cavity inject the anaesthesia, then with 4% multitude gather formal dehyde to left ventricles. Fixed, packed, cut into slices, applied SABC method. The first antibody which is the bcl - 2 and bFGF IgG of rat antibody multitude . The second antibody is a biotinise goat anti - rat. DABto show the colour, normal regulations to dehydrate, transparent, sealing slices. Each specimen took two slices ,the difference was random to observe 8 visual fields in the hippocampus area. Final,we detect the expression of the bFGF and bcl - 2 after24,48 and 72 hours of cerebral ischemic reperfusion by microimage analysis system and immunohistochemistry. 3. Statistics the method ;Data with all the mean ± Sigma (x ± s) , the application SPSSV11. 5 software carried on the processing, the adoption t examine, p < 0. 05 think that the covariance learns the meaning.ResultInfarct volumes test; comparing the control group with plasmid control group,there is no distinctive difference in the different temporal point(p > 0. 05 ). Comparing the bcl - 2 group with control group, infarct volumes reduced obviously in the different temporal point (p <0.05). Expression of bcl -2 protein-.no obvious difference between the control group and plasmid control group at each temporal point after infarct( p > 0.05 ). Comparing the bcl - 2 group with the former two groups, the bcl - 2 protein expression of bcl - 2 group at each temporal point after lack of blood increased obviously ( p <0.005 ). Expression of bFGF protein;no obvious difference between the controlgroup and plasmid control group at each temporal point ( p > 0. 05 ). Comparing with the former two groups, the bFGF expression of the bcl - 2 group at each temporal point increased strikingly.ConclusionWe injected pLXSN - bcl -2 plasmid into the rats' internal carotid arteryafter temporary ischemia of the middle cerebral artery occlusion ( MCAO) for 2 hours, the infarct volumes reduced obviously. The result of immunohistochemistry shows: the expression of bcl - 2 protein and bFGF albumen increased obviously at each temporal point after lack of blood. Therefore, we conclude that bcl - 2 gene can protect nerve cell and its brain , decrease the infarct volumes and it is possible that bcl - 2 gene promotes the expression of bFGF.