| Objective:Our study was to observe the expression of brain-derived neurotrophic factor(BDNF)and various mRNA transcripts in different regions of hippocampus after cerebral ischemia,and to lay a foundation for subsequent histone acetylation modification,and to open up a new way for treatment brain injury.Methods:Wistar male rats weighing 180-250 g were divided into control group 、 sham operation group and ischemia model group(6h 、24h、48h three groups)randomly.Cage rearing in each group of rats and 2 rats per cage.These rats were housed under light/dark 12h/12 h and have an access to water and food.All animals were allowed to adjust to new conditions for 7 days.Rats were fasted overnight and anesthetized with chloral hydrate(40mg/100 g.i.p),then the rats were were fixed on the console.Along the first cervical vertebra oblique nerve to find the wing protruding hole.Bilateral vertebral artery below the pinhole were electro-coagulated with a shape slender,and sutured the skin.The common carotid arteries were separated,thread and suture the skin,and rats were fasted at night.The next day,the commom carotid artery was clamped for 15 min,resulting in transient forebrain ishemic.In the sham operation group,except the non-occlusion vertebral artery and the commom carotid artery,the other operations were the same as the model group.The control group did not do any treatment.After making the model,rats randomly were selected from five groups totake the brains off the head,and divided the hippocampus into CA1 and CA3 regions for conducting Western Blot and RT-PCR methods to detect the expression levels of BDNF protein and BDNF mRNA.The other rats were cultured with 7d post-perfusion section for niston-staining,and changes of pyramidal neurons in CA1 and CA3 region.were observed under the microscope.Results:1.Nissl staining showed that neurons in hippocampal CA1 region of rats in control group and sham operation group were arranged closely and neatly,with normal structure and regular nuclei.The number of normal neurons in hippocampal CA1 area of ischemic model group was significantly reduced.A large number of pyramidal neurons degenerated and necrotic,the nucleus was irregular and contracted deeply,and the intercellular space was significantly increased.2.qPCR results suggested that the expression of BDNF mRNA I and BDNF mRNA IV were increased 6 h group in the CA3 area after ischemia compared with the expression of the sham group(n = 3,P<0.05),and BDNF mRNA VIII was increased significantly(n = 3,P<0.01).The expression of BDNF mRNA IIb was significantly increased 48 h group in the CA3 area after ischemia(n = 3,P < 0.05),and the expression of BDNF mRNA VI in the 6h group of CA3 area was significantly increased after ischemia(n = 3,P< 0.05).The expression of BDNF mRNA IIa,BDNF mRNA IIc,BDNF mRNA III,BDNF mRNA V,BDNF mRNA VIIand BDNF m RNA IX had no change.3.The results of Western Blot showed that there was no statistically significant difference in the expression of BDNF protein between CA1 area of the model groups.Compared with the normal group and sham group,the expression of BDNF in CA1 area was decreased significantly,there is statistical significance(P < 0.05,n = 3).The expression of BDNF in model group rats in CA3 of hippocampal area was no statistical difference compared with the rats of the normal group and sham group.Conclusion:1.The cerebral ischemia model of rats was successfully established by using the modified Pulsinelli four-vessel method.2.There were different BDNF mRNA transcripts in the CA1 region and CA3 region of the hippocampus of the ischemia model group rats.3.BDNF protein expression was associated with different transcription of BDNF mRNA. |
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