| PrefaceApoptosis appears to contribute to posischemic neuronal death in stroke and the expression of Bcl - 2 gene has been shown to rescue neuronal cells from pro-grammed cell death. It is known that TGF - β1, played a neuroprotective role. How about the connection between Bcl -2 and TGF -β1 As well as, the plas-mid pLXSN was increasingly focused on for its safety and effect. So we established temporarily middle cerebral artery occlusion ( MCAO) in rat and studied whether Bdl -2 can improve cerebral infarct and expression of TGF - β1 by in-traventricular injection.Methods135 wister rat were randomly assigned in 3 groups; Bcl -2 - treated group, saline group, pLXSN group, there were 45 rats in each group. Firstly, temporary ischemia of the middle cerebral artery occlusion (MC AO) for 2 hours was induced by the suture occlusion techique. Secondly, we injected saline, pLXSN plasmid and pLXSN - Bcl -2 plasmid into the rats'lateral ventricle soon after infarction. Finally, according to MCAO model above, the lateral ventricle was injected in rats successfully, we measured the following things. (1) The cerebral infarction volume: We detected the cerebral infarction volume at 3 , 6, 24, 48, 72 hours after cerebral ischemic reperfusion by TTC staining, 4% multitude gather formaldehyde and image analysis system. (2 ) immunohistochemistry of Bcl -2 and TGF -β1 10% the water match chlorine aldehyde, 0. 35ml/100gbelly carities injected the anaesthesia, then with 4% multitude gather formaldehyde to left ventricles. Fixed,packed, cut into slices, applied SABC method. The first antibody which was Bcl - 2 IgG of rat antibody ( Ab - 1) and TGF - β1 IgG of rat antibody multitude clone ( bought from Wuhan Baster Company). The second antibody was biotinise goat anti - rat. DAB to show the colour, normal regulations to dehydrate, transparent,sealing shices. Finally, we detected the expression of TGF - β1 and Bcl - 2 after 3 , 6, 24, 48, 72hours of cerebral is-chemic reperfusion by microimage analysis system and immunohistochemistry.ResultsThe results were: (1) The expression of Bcl - 2 protein was detected by microimage analysis system and immunohistochemistry; The expression of Bcl -2 protein of the saline group and the pLXSN group were not different (P >0. 05 ). The expression of the Bcl - 2 protein of Bcl - 2 group was more than the other two groups at the same time point ( P < 0. 05 ) , and it achieved the peak value at the 24th. The expression of Bcl -2 mascul cells were presented at pec-tin cell liquid. (2) The expression of TGF - β1, was detected by microimage a-nalysis system and immunohistochemistry: The expression of TGF - β1 of the saline group and the pLXSN group were not different (P >0. 05). The expression of TGF - β1 of the Bcl -2 group was more than the other two groups at the same time point (P <0.05) ,and it achieved the peak value at the 6th and 48th. The positive expression cells were mainly in the verge of ischemia, to be presented at pectin cell liquid. (3) The cerebral infarction volume of different reperfusion time point which was detected by image analysis system and TTC staining: The cerebral infarction volume of the saline group and the pLXSN group was not different ( P > 0. 05 ). The cerebral infarction volume of the Bcl - 2 group was less than the saline group and the pLXSN group in 24h, 48h,'72h after MCAO (P < 0. 05) , there were not different between three groups in 3h and 6h after MCAO ( P > 0.05). The result of statistics of every group is showed with x s, using T test. Result the P is worth all smaller 0.05 and has the statistics meaning.ConclusionOur study demonstrated Bcl - 2 could decrease cerebral infarction volume of the rat and increase the expression of TGF - β1 by intraventricular injection, and it could protect the cerebral tissue against ischemic injury. Accordingly, this method of transgenic therapy was feasible for rat transient focal cerebral ischemia , One possible mechanism of action of Bcl - 2 after temporary ischemia appeared to be... |
