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Objective: In this study, using in vitro recombinant techniques, preparation pcDNA3.1-HIF-1α plasmid. Injected into the HIF-1α plasmid via a cervical neck - the internal carotid artery, middle cerebral artery ischemia-reperfusion injury in the rat brain, 6h, 12h, 24h, 72h and 7d, take the rat brain tissue glial the subtypes cell line specific immunohistochemical staining groups subtypes cells was observed. Fluorescent double staining of HIF-1α, VEGF same the glial subtypes cells line in the laser scanning confocal microscope to observe the co-expression. Comprehensive Evaluation of the above experimental results recombinant plasmid for the therapeutic effect of cerebral infarction, and its remodeling regulatory mechanism on a variety of cells to repair of HIF-1α. Methods: HIF-1α in vitro cDNA sequencing to detect Houkekelong into the eukaryotic expression vector pcDNA3.1 enzyme digestion recombinants rat brain artery cerebral ischemia and reperfusion model making reference to Zea Longa et al reported the method and make improvements. 10% chloral hydrate intraperitoneal anesthesia, neck midline incision, separation of muscle tissue exposed neck vessels, insert line tied to the internal carotid artery through the external carotid artery, the the line bolt head end from the blood vessels at the bifurcation 1.8cm so as intraluminal suture infarct artery in the brain. 2h after the withdrawal string tied injected plasmid of HIF-1α specimens 6h, 12h, 24h, 72h and 7d observed in the control group injected PBS; corresponding point in time the experimental group and the control group rats were killed and brains were removed after the optic chiasm after 2mm thick serial sections, the first piece to do TTC staining, the rest do paraffin-embedded, 5μm thick paraffin sections, HE staining and immunohistochemical staining of glial subtypes cell line specific, high magnification were counted different point in time the experimental group and the number of of glial subtypes cells in the control group; 4 paraffin sections of HIF-1α, VEGF with glial subtypes cells do double immunofluorescent staining in laser scanning confocal microscope to observe the positive The overlap of the cells, glial subtypes cells exists with the expression of HIF-1α, VEGF; Statistical analysis: The experimental data were statistically analyzed using the SPSS15.0 statistical software. Measurement data between the two groups compared using t-test, P lt; 0.05 indicates difference was statistically significant; Results: RT-PCR amplification of HIF-1αcDNA sequencing results with Genebank record exactly the same build and restriction endonuclease pcDNA3 .1-HIF-1α plasmid; 2 middle cerebral artery cerebral ischemia-reperfusion injury through the external carotid artery injection of HIF-1α plasmid group and PBS group mortality was no significant difference, but TTC staining after 72h treatment group infarct area was significantly less than the control group, and survival 7d rats neurological deficit scores were significantly lower than the PBS group; 3 HIF-1α on cerebral ischemia reperfusion subtypes of glial cells: rat brain ischemia-reperfusion 6h, 12h, 24h, 72h and 7d, the number of astrocytes (GFAP-positive cells) by the external carotid artery injection of HIF-1α plasmid group mean were 12.20 ± 2.864,18.80 ± 4.207,30.00 ±, 4.301,38.40 ± 2.881 and 43.20 ± 6.978, the number of injections of PBS group mean of 15.80 ± 4.494,18.60 ± 1.673,23.00 ± 4.583,31.60 ± 3.361 and 29.00 ± 6.000, both positive cells of the same point in time mean to do t test compared with ischemia-reperfusion injury 24h and subsequent time points, HIF-1α and control groups differences (P lt; 0.05) in the number of positive cells; the carotid injection HIF-1α plasmid microglia (Isolectin- B4-positive cells) were 8.400 ± 2.074,14.40 ± 4.393,20.80 ± 2.280,17.00 ± 3.162 and 11.60 ± 2.408, PBS control group positive cells mean 8.400 ± 2.302,15.40 ± 2.402,25.80 ± 4.147,37.80 ± mean 6.181 and 24.80 ± 4.325, two sets of positive cells of the same point in time mean to do t test compared with ischemia-reperfusion injury in 24h and after each point in time there are differences between the two groups was statistically significant (P lt; 0.05). HIF-1α injection group oligodendrocytes (CNPase positive cells) are, respectively, PBS control group the average number of positive cells were 16.60 ± 17.00 ± 2.916,11.40 ± 2.302,10.00 ± 3.526,16.40 ± 2.302 and 19.20 ± 2.388 2.408,10.60 ± 2.074,7.200 ± 1.924,4.000 ± 1.581 and 13.20 ± 2.388, both positive cells of the same point in time mean to do t test comparing ischemia-reperfusion injury in 72h between the two groups, a statistically significant difference in the number of positive cells ; 4 HIF-1α, VEGF and glial subtypes cells co-expressed: paraffin sections double immunofluorescent staining in laser confocal observed under a microscope and found that a large number of astrocytes (GFAP-positive cells) and HIF-1α The co-expression of VEGF protein, no microglia (Isolectin-B4-positive cells) and oligodendrocytes (CNPase positive cells) in HIF-1α, VEGF obvious expression. Conclusion: The external carotid - internal carotid artery injection of recombinant pcDNA3.1-HIF-1α plasmid can effectively reduce infarct size in rats with cerebral ischemia-reperfusion injury in brain tissue. 2 rat brain after ischemia-reperfusion injury through external jugular - internal carotid arterial injection of recombinant pcDNA3.1-of HIF-1α plasmid can promote increased astrocytes, oligodendrocytes increase in the number of glial cells, and effectively suppress small plastic proliferation of interstitial cells, play a neuroprotective effect. 3 rat cerebral ischemia-reperfusion injury, glial subtypes cells, HIF-1α protein is mainly expressed in astrocytes.
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