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

To investigate the expression of bFGF in rat with bcl-2 gene injectived after the MCAO for 2 hours was induced and to study the protective effect of bcl-2 a-gainst neuroal injury and the possible roles of alteration in the expression of bFGF following cerebral ischemia in rat.Methods150 wistar rats were randomly assigned in 3 groups: bcl-2-treated group, saline group, pLXSN group, there were 50 rats in each group. Firstly, temporary ischemia of the middle cerebral artery occlusion ( MCAO) for 2 hours was induced by the suture occlusion technique. Secondly, we injected saline, pLXSN plasmid and pLXSN-bcl-2 plasmid into the rats' lateral ventricle soon after infarction. Finally, according to the methods above-mentioned to make MCAO model and the lateral ventricle injected rats successfully, we measure these following things. (1) The cerebral infarction volume: We detect the cerebral infarction volume after 3,6,24,48 and 72 hours of cerebral ischemic reperfusion by TTC staining ,4% multitude gather formaldehyde and image analysis system. (2) immunohistochemistry of bcl-2 and bFGF: 10% the water match chlorine aldehyde, 3. 5ml/kg belly carities 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 (Ab-1) and bFGF IgG of rat antibody multitude ( bought from Wuhan Baster Company). The secend antibody is a biotinise goat anti-rat. DABto show the colour, normal regulations to dehydrate, transparent, sealing shices. Finally, we detect the expression of the bFGF and bcl-2 after 3 , 6, 24, 48, 72 hours of cerebral ischemic reperfusion by microimage analysis system and immu-nohistochemistry.ResultInfarct volumes test: comparing the saline group with pLXSN group, there is no distinctive difference in the different temporal point. Comparing with the treated group and contrasted group, no striking difference at 3-hour and 6-hour points(p > 0. 05 ) , reducing obviously at 24,48,72 hour( p < 0. 05 ). Expression of bcl-2 protein; no obvious difference between the saline group and pLXSN group, at each temporal point after infarct (p > 0. 05) , the result showed the peak point of expression is the sixth hour after losing blood. Comparing the experiment group (inject pLXSN-bcl-2 in the cerebral ventricle ) with the former two groups, the expression of each temporal point after lack of blood increased obviously (p <0. 005) ,the peak point of expression is the 24th hour after lacking of blood. Expression of bFGF protein: no obvious difference between the saline group and pLXSN group, the expression is no strinking difference at each temporal point after lacking of blood (p > 0. 05) ,the result showed the peak point of expression is the 24th hour after lacking of blood, which agrees with literature report. Comparing with pLXSN group, the expression of the bcl-2 group at each temporal point increased strikingly (p <0. 05) ,the peak point of expression is the 3 th hour after lacking of blood and continued to 72th hour.ConclusionWe injected pLXSN-bcl-2 plasmid into the rats' lateral ventricle soon after temporary ischemia of the middle cerebral artery occlusion ( MCAO) for 2 hours, the infarct volumes reduced obviously at 24,48,72 hour( p < 0.05 ). The effect result of this experiment shows: comparing the experiment group (inject pLXSN -bcl-2 in the cerebral ventricle) with the former two groups, the expressionof each temporal point after lack of blood increased obviously (p <0. 005) ,the peak point of expression is the 24th hour after lacking of blood. BFGF albumen in bcl-2 groups is obviously better than two contrast groups in dififerent time,moreover, obviously up-regulated within 3 hours until continuously goes to 72 hours. Conclusionally, bcl-2 gene promotes bFGF expression, it protects nerve cell and nutrientcoUoid cell and promote new blood vessel to form etc, therefore it protects its brain.