本文選題：豐富　切入點：環(huán)境　出處：《復旦大學》2014年碩士論文

【摘要】：目的：既往研究證實,缺血性腦梗塞后的血流恢復及血管再生對腦功能的改善有重要作用。本實驗擬探討豐富環(huán)境干預是否能改善腦卒中后血管新生和腦血流的改善。方法：將SD大鼠隨機分為3組：假手術組、豐富環(huán)境組和標準環(huán)境組,并進行大腦中動脈梗塞手術,其中假手術組除不堵塞血管這一步驟,其余步驟與手術組相同。在缺血前和缺血1小時分別運用激光散斑技術檢測各組大鼠大腦中動脈供血區(qū)皮層的相對血流速度,然后將手術后的大鼠分別放入相應的環(huán)境中飼養(yǎng)4周,4周后再次檢測同一區(qū)域皮層相對血流速度。另有大鼠干預4周后經大鼠尾靜脈注入異硫氰酸熒光素右旋糖苷(Fluorescein Isothiocyanate dextran, FITC-dextran),5分鐘后斷頭取腦,固定后切片,通過激光共聚焦掃描顯微鏡檢測缺血干預后的血管密度。同時,運用免疫印記(Western blot)技術檢測缺血側大腦皮層血管內皮生長因子(Vascular Endothelial Growth Factor, VEGF)的表達。結果：與缺血前比較,豐富環(huán)境組和標準環(huán)境組缺血側皮層的相對血流速度在缺血1小時后明顯下降(P0.01),假手術組無明顯變化。干預4周后,與標準環(huán)境組相比,豐富環(huán)境組的缺血側皮層血流速度明顯增加(P0.05)。另外,豐富環(huán)境干預4周后的大鼠缺血側皮層的血管密度明顯大于標準環(huán)境組,VEGF蛋白表達也明顯高于標準環(huán)境組。結論：豐富環(huán)境可以增加缺血后大鼠的缺血側皮層血流,血流的增加可能與豐富環(huán)境促進了VEGF的表達從而增強血管再生有關。目的：血管新生是成人腦缺血后血管生成的最主要方式,也是現(xiàn)如今研究的主要方向；腦微血管內皮細胞是主要的腦血管組成成分,對腦缺血后的血管新生及其重要。本項研究將探討豐富環(huán)境干預后大鼠的血清對氧糖剝奪后腦微血管內皮細胞的增殖作用。方法：豐富環(huán)境及標準環(huán)境干預同前,干預后大鼠心臟采血,離心后取上清,過濾后備用。培養(yǎng)腦微血管內皮細胞,實施氧糖剝奪實驗以復制體外腦缺血模型。而后進行預實驗,用10%、20%和40%濃度的不同干預血清分別培養(yǎng)氧糖剝奪后的腦微血管內皮細胞,用CCK-8測定其增殖,摸索最適濃度。最后,用最適濃度血清多次重復培養(yǎng)氧糖剝奪后的腦微血管內皮細胞,進一步探討豐富環(huán)境干預后大鼠的血清對氧糖剝奪后的腦微血管內皮細胞的增殖作用。結果：10%濃度的干預血清培養(yǎng)氧糖剝奪后的腦微血管內皮細胞,其CCK-8檢測的OD值明顯大于20%和40%濃度的干預血清(P0.01)；10%濃度豐富環(huán)境干預的大鼠血清培養(yǎng)氧糖剝奪后的腦微血管內皮細胞,其CCK-8檢測的OD值明顯大于標準環(huán)境干預的大鼠血清的CCK-8值(P0.01)。結論：10%濃度的干預血清對氧糖剝奪后的腦微血管內皮細胞的促增殖作用最明顯；10%濃度豐富環(huán)境干預的大鼠血清對氧糖剝奪后的腦微血管內皮細胞的促增殖作用明顯優(yōu)于同濃度標準環(huán)境干預的大鼠血清。
[Abstract]:Objective: past studies have confirmed that, Blood flow recovery and vascular regeneration after ischemic cerebral infarction play an important role in the improvement of cerebral function. The purpose of this study was to explore whether rich environment intervention can improve angiogenesis and cerebral blood flow after stroke in SD rats. They were randomly divided into three groups: sham-operated group, Enrich the environment group and the standard environment group, and perform the middle cerebral artery infarction operation, the sham-operation group except the step of not blocking the blood vessel, The other steps were the same as those in the operation group. Laser speckle technique was used to detect the relative blood flow velocity in the cortex of the middle cerebral artery in each group before and after 1 hour of ischemia. Then the rats were fed in the corresponding environment for 4 weeks and 4 weeks later, the relative blood flow velocity of cortex in the same area was measured again. After 4 weeks of intervention, the dextrose of fluorescein isothiocyanate was injected into the caudal vein of the rats. Fluorescein Isothiocyanate dextran, FITC-dextranan, and brain were taken after 5 minutes. After fixation, the vascular density was detected by laser confocal scanning microscope. At the same time, The expression of vascular Endothelial Growth factor (VEGF) in ischemic cerebral cortex was detected by Western blot technique. The relative blood flow velocity of ischemic cortex in the rich environment group and the standard environment group decreased significantly after 1 hour of ischemia, but there was no significant change in the sham operation group. After 4 weeks of intervention, compared with the standard environment group, the relative blood flow velocity of the ischemic cortex decreased significantly in the rich environment group and the standard environment group. In the rich environment group, the blood flow velocity of ischemic cortex increased significantly (P 0.05). The vascular density of ischemic cortex of rats after 4 weeks of rich environment intervention was significantly higher than that of standard environment group, and the expression of VEGF protein was also significantly higher than that of standard environment group. Conclusion: rich environment can increase the blood flow of ischemic side cortex of rats after ischemia. The increase of blood flow may be related to the enhanced expression of VEGF and the enhancement of angiogenesis. Objective: angiogenesis is the most important way of angiogenesis after cerebral ischemia in adults, and it is also the main research direction nowadays. Cerebral microvascular endothelial cells are the main components of cerebral vessels. This study will explore the effect of serum enriched environment intervention on the proliferation of cerebral microvascular endothelial cells after oxygen glucose deprivation in rats. After intervention, the rat heart blood was collected, the supernatant was taken after centrifugation, and then filtered. The cerebral microvascular endothelial cells were cultured, and the oxygen glucose deprivation test was carried out to replicate the model of cerebral ischemia in vitro. Brain microvascular endothelial cells were cultured with different concentrations of 10% and 40% different concentrations of oxygen glucose deprivation, and their proliferation was determined by CCK-8, and the optimal concentration was found. Brain microvascular endothelial cells were cultured repeatedly with the optimal concentration of serum for several times after oxygen glucose deprivation. To further study the proliferation effect of serum of rats exposed to enriched environment on cerebral microvascular endothelial cells after oxygen glucose deprivation. Results the cultured cerebral microvascular endothelial cells were cultured with 10% concentration of 10% concentration of intervention serum. The OD value detected by CCK-8 was significantly higher than that of 20% and 40% of the intervention serum P0.01T10%, which enriched the concentration of cultured cerebral microvascular endothelial cells after oxygen glucose deprivation in rats. The OD value detected by CCK-8 was significantly higher than that of serum CCK-8 of rats treated with standard environmental intervention (P 0.01). Conclusion the concentration of 10% 10% of the intervention serum can promote the proliferation of cerebral microvascular endothelial cells after oxygen glucose deprivation, and the concentration of 10% can enrich the concentration of environmental intervention. The proliferative effect of rat serum on cerebral microvascular endothelial cells after oxygen glucose deprivation was significantly better than that of rats treated with the same concentration of standard environment.
【學位授予單位】：復旦大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R743

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