本文選題：缺血性卒中 + 內(nèi)皮祖細(xì)胞��； 參考：《蘇州大學(xué)》2014年碩士論文

【摘要】：目的：觀察不同劑量丁苯酞對(duì)大腦中動(dòng)脈缺血（MCAO）/再灌注模型大鼠神經(jīng)功能、循環(huán)內(nèi)皮祖細(xì)胞(circulating endothelial progenitor cells, EPCs)水平及梗死區(qū)血管新生的影響，探討其治療缺血性卒中的療效及機(jī)制。 方法：選取健康雄性大鼠Sprague-Dawley（SD）大鼠48只，體重（250±20）g， 按照隨機(jī)分組法分為假手術(shù)組，模型對(duì)照組，NBP低劑量治療組（60mg/kg.d）和NBP高劑量治療組(120mg/kg.d），每組各12只。假手術(shù)組僅分離頸部血管而不插入線栓。其他三組均采用線栓法建立大鼠MCAO/再灌注模型，假手術(shù)組與模型組各取2只行TTC染色法觀察大鼠梗塞區(qū)形態(tài)學(xué)改變，其余大鼠分別給予安慰劑(大豆油)（2ml/d）及兩種劑量的NBP灌胃治療，，持續(xù)5d。每天對(duì)四組大鼠進(jìn)行神經(jīng)功能缺損評(píng)分；給藥前及灌胃5d后分兩次采集大鼠外周血，流式細(xì)胞儀檢測(cè)治療前后循環(huán)EPCs的數(shù)量；灌胃5d后行血管內(nèi)皮細(xì)胞標(biāo)記物CD31及vWF免疫組化染色，觀察各組大鼠缺血區(qū)血管新生情況。GraphPad5.0軟件系統(tǒng)對(duì)實(shí)驗(yàn)數(shù)據(jù)進(jìn)行統(tǒng)計(jì)分析處理。 結(jié)果： 1）腦TTC染色后可見線栓側(cè)大腦半球明顯梗死病灶，呈蒼白色，非梗死區(qū)顯示為紅色； 2）NBP治療第1d，低劑量組大鼠神經(jīng)功能評(píng)分開始出現(xiàn)降低（P0.05），高劑量組神經(jīng)功能評(píng)分明顯降低（P0.01），模型對(duì)照組大鼠神經(jīng)功能評(píng)分在灌胃3d后出現(xiàn)明顯降低（P0.01）； 3）NBP治療第1d，低劑量組大鼠神經(jīng)功能評(píng)分低于對(duì)照組（P0.05），高劑量組神經(jīng)功能評(píng)分明顯低于對(duì)照組（P0.01）；第2-5d，NBP兩種劑量治療組大鼠神經(jīng)功能評(píng)分均顯著低于對(duì)照組（P0.01）； 4）NBP治療第3d，高劑量組神經(jīng)功能評(píng)分明顯低于低劑量組（P0.01）； 5）灌胃5d后與缺血2h再灌注時(shí)比較，假手術(shù)組大鼠外周血EPCs數(shù)量無明顯變化，其余各組均顯著增加（P0.01）； 6）不同劑量NBP治療組大鼠在5d后的外周血EPCs數(shù)量均顯著高于對(duì)照組（P＜0.01）；NBP高劑量組外周血EPCs水平顯著高于低劑量組（P0.01）； 7）假手術(shù)組腦切片顯微鏡下可見抗CD31陽(yáng)性細(xì)胞和抗vWF陽(yáng)性細(xì)胞，呈正常腦組織內(nèi)血管染色；對(duì)照組腦切片幾乎未見或極少量表達(dá)抗CD31陽(yáng)性或抗vWF陽(yáng)性細(xì)胞，提示腦缺血區(qū)血管破壞；NBP治療組腦切片在光鏡下均可看到抗CD31陽(yáng)性及抗vWF陽(yáng)性細(xì)胞大量表達(dá)，其中NBP高劑量治療組的陽(yáng)性細(xì)胞表達(dá)更為顯著。 結(jié)論： 1）MCAO/再灌注損傷可促進(jìn)EPCs動(dòng)員； 2）丁苯酞可進(jìn)一步增強(qiáng)循環(huán)EPCs動(dòng)員，有效地促進(jìn)缺血區(qū)血管新生，改善神經(jīng)功能缺損評(píng)分；高劑量較低劑量療效更顯著。
[Abstract]:Objective: to observe the effects of different doses of butyphthalide on nerve function, circulating endothelial progenitor cell (endothelial progenitor cells, EPCs) level and angiogenesis in infarcted area of rats with middle cerebral artery ischemia (MCAO) / reperfusion, and to explore the therapeutic effect and mechanism of butyphthalide on ischemic stroke. Methods: a total of 48 healthy male Sprague-Dawley SD rats were selected with a body weight of 250 鹵20g. According to the method of random grouping, the model group was divided into sham operation group, model control group, low dose treatment group (60 mg / kg 路d) and NBP high dose group (120 mg / kg 路dg / d), with 12 rats in each group. In the sham operation group, only cervical vessels were separated and no thread embolus was inserted. The other three groups were used to establish MCAO/ reperfusion model in rats. Two rats in sham-operation group and two in model group were taken to observe the morphological changes of infarct area by TTC staining. The rest of the rats were given placebo (soybean oil 2 ml / d) and two doses of NBP for 5 days. The peripheral blood was collected twice before and after 5 days of administration, and the number of circulating EPCs before and after treatment was detected by flow cytometry. After 5 days of gastric perfusion, the vascular endothelial cell markers (CD31 and vWF) were stained by immunohistochemistry. The angiogenesis in ischemic area of rats in each group was observed. The experimental data were statistically analyzed by GraphPad 5.0 software system. Results: 1) after TTC staining, the cerebral hemispheres showed obvious infarct lesions, which were pale and red in non-infarcted areas. On the 1st day after 2)NBP treatment, the neurological function scores of the rats in the low dose group began to decrease P0.05A, in the high dose group decreased P0.01a significantly, and in the model control group, the neurological function scores of the rats in the model control group decreased significantly after 3 days of gastric administration. On the 1st day of 3)NBP treatment, the scores of nerve function in the low dose group were lower than those in the control group (P 0.05), the scores in the high dose group were significantly lower than those in the control group (P 0.01), and the scores of nerve function in the 2 to 5 days treatment group were significantly lower than those in the control group (P 0.01). On the 3rd day after 4)NBP treatment, the scores of nerve function in the high dose group were significantly lower than those in the low dose group (P 0.01). 5) after 5 days of gastric perfusion, there was no significant change in the number of EPCs in peripheral blood of rats in sham-operated group compared with that at 2 h after ischemia and reperfusion. 6) the number of EPCs in peripheral blood of rats treated with different doses of NBP after 5 days was significantly higher than that of control group (P < 0.01). The level of EPCs in high dose group was significantly higher than that in low-dose group (P 0.01). 7) Anti CD31 positive cells and anti vWF positive cells were observed under microscope in sham-operation group, and showed vascular staining in normal brain tissue, while no or very few anti CD31 positive or anti vWF positive cells were found in brain sections of the control group. It was suggested that the positive cells of anti- and anti- could be seen in the brain sections of the cerebral ischemic area treated with NBP under light microscope, especially in the high dose group of NBP. The results showed that the expression of anti-NBP positive cells was more significant in the high-dose NBP group than in the high-dose NBP group. Conclusion: 1)MCAO/ reperfusion injury can promote EPCs mobilization. 2) Butylphthalide could further enhance the mobilization of circulating EPCs, promote angiogenesis in ischemic area and improve the score of nerve function defect, and the effect of high dose was more obvious than that of low dose.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R743.3

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