The Study Of VEGF Expressions Of Grafting Bone Marrow Stromal Cell Associated With Bcl-2 Gene For Treating Ischemic Braim In Rats

PrefaceBone marrow stromal cell (BMSC) is a source of neuro-tranplantation. BMSCs can differentiate into neurous and release of trophicfacetors such as VEGF, GDNEM any experiements have proved that BMSCs is effective on improvement of neurological functional deficits after stroke in rats.Bcl-2 gene is the first gene that has been acknowaged to have the function of suppress eukaryoncell apoptosis. Bcl-2 is also effection on improvement of neurological unctional deficits after stroke in rats.VEGF is the most important tyophic factors that facilitate repair of damage brain. The expression VEGF can be improved if the brain was damaged.Experiments maded by Yi Li. Chen eat has proved that. Coadministration of BMSCs with a cell permeable inhibitor of caspases, Z-Val-Ala-b-Asp-fluovomethylke tone (Z-VAD), into the MCAo rats promofes BMSCs survival and improve cutcome.We tested uhether Coadminis-tration of BMSCs with bcl-2 into the MCAO rats improves functional recovery after stroke and observe VEGF exorssion.Materids and Methods40 adult male wistar rats were assigned randomly into 4 groups:nothing group (n=10),BMSCs group (n=10) bcl-2 group and BMSLs with bcl-2 group (n=10).Then each group was divided into 2 subgroups:3 days,14 days,according to the time after reperfusion onset.Thereafter, middle cerebral artery acclusion (MCAO) was performed with the help of intraluminal thread to each rat.Tow hours after the ischemia insult, reperfusion was implemented to those rats. Then three hours later, plxsn -bcl-2 was respectively injected into bel-2 group and BMSCs with bcl-2 group through interial carortid artery.After BNSCs were proliferated,marked with Brcdu and pro-diferentiated into vitro. The Brcdu-marked BMSCs were injected into BMSCs group and BMSCs with bcl-2 group through til vein after 24 hours reperfusion.Immunohis to chemistry determination and TUND techniques: the rats in each subgroup were anaesthetized again immediately after that,transcardiac perfusion was performed to each of them.Then the brains were removed,postfixed,embedded in paraffin and consecutively sectioned (6μm thick covonal sections). The sections were subject to staining procedures after deparaffination. Using SABC Immunohistochemistry kit to observe bax.. vEGF BMSCs distribution and TUNel techniques to check out the distribution of apoptosis neurous.Determination of the result: Taken two sections of each sample,examined and photographed usingComputerized video imaging microscopy. Date were acquired from diffirent eight fields in corpus striatum region by analysing the positive area. percentage at high power magnification(400X).All the results were transformed to dates through image analysis system ,which were expressed as x±s.the values were analyzed,using ANOVA and test of spss software. A value of p<0.05 was lonsidered significant, obvious significance was set at p<0.01.ResultsThe result of bax technigues was that the levels of bax in BMSCs with bcl—2 group was greatly attenuated compared with that of BMSCs group, bcl—2group and nothing group. The levels of bax in BMSCs group, or bcl—2 group was greatly attenuated compared with that of nothing group.The elevated levels of VEGF were significantly attenuated in BMSCs with bcl-2 group as compared to the other groups(p<0.05＝,The elevated levelsof VEGF in BMSCs group or bcl-2group was greatly attenuated compared with that of nothing group.The result of TUNEL technigues was that the number of apoptosis neurous in BMSCs with bcl-2 group reduced greatly compared with that of control groups(p< 0.05＝.the number of cepoptosis neurous in BMSCs group or bcl-2 group was greatly attenuated compared with that of nothing group.The majority of Brdu-labeded BMSCs were located in the ischemic boundary zone.A few brdu-labeled BMSCs were located in the normal sphare.ConclusionAt the time of I/R injury, coadministration of bone marrow cells (bmscs) with bcl-2 could enhance the expression of VEGF, improve the nerological function combing treatment could provide a potentiol therapertic strategies to the treatment of ischemic stroke.