本文選題：毛冬青甲素　切入點：局灶性腦缺血再灌注　出處：《福建醫(yī)科大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：研究毛冬青甲素（Ilexonin A，IA）對大鼠腦缺血再灌注后缺血周邊區(qū)Notch信號的調(diào)節(jié)作用與神經(jīng)干細(xì)胞增殖和分化的關(guān)系，，探討腦損傷后的修復(fù)機制。 方法:線栓左側(cè)大腦中動脈阻塞（middle cerebral artery occlusion，MCAO）2h，制備腦缺血再灌注損傷模型。 SD大鼠隨機分為：正常組、假手術(shù)組、模型組、IA組，其中模型組、IA組（IA40mg/kg）分別再灌注不同時間。神經(jīng)功能學(xué)評分評估大鼠的神經(jīng)功能恢復(fù)情況；TTC染色驗證實驗動物模型和提供定位標(biāo)記；免疫熒光法觀察缺血周邊區(qū)增值細(xì)胞標(biāo)記物Brdu、神經(jīng)干細(xì)胞標(biāo)志物巢蛋白（Nestin）共表達的陽性細(xì)胞數(shù)，Brdu、神經(jīng)元標(biāo)志物神經(jīng)元核蛋白（Neuronalnucleus protein,Neun）共表達的陽性細(xì)胞數(shù)及Notch信號通路分子Notch信號胞內(nèi)域(Notch intracellular domain,NICD)的陽性細(xì)胞數(shù)，蛋白印跡法檢測NICD的表達情況；RT-PCR法檢測Notch信號通路中Notch1受體蛋白、早老蛋白1（Presenilin,PS1）、靶基因HES1的表達情況。 結(jié)果：（1）神經(jīng)功能恢復(fù)情況：模型組和IA組腦缺血再灌注1天時神經(jīng)功能缺損評分明顯增高，隨著時間的增加評分逐漸降低，并且IA組與模型組在3天、7天、14天降低顯著（p0.01、p0.05）。（2）缺血周邊區(qū)神經(jīng)再生情況：IA組與模型組在再灌注第3天相比較，IA組缺血周邊區(qū)Brdu/Nestin陽性細(xì)胞數(shù)增殖明顯（p0.05），7天時表達的細(xì)胞數(shù)最多（p0.05）。再灌注第14天，IA組和模型組Brdu/Neun陽性細(xì)胞僅有少量表達且無顯著性意義。（3）Notch信號通路的激活和各基因的表達情況：模型組和IA組與正常組和假手術(shù)組比較，在再灌注1天NICD表達增加，且在1天、3天、7天均增高（p0.05）。模型組、IA組Notch信號通路分子Notch1, PS1, HES1的表達較正常組和假手術(shù)組均升高，IA組與模型組比較在3天，7天時表達增加（p0.01）。 結(jié)論 1. IA在腦缺血再灌注后促進大鼠神經(jīng)功能的恢復(fù)，其機制與IA促進缺血周邊區(qū)Nestin陽性細(xì)胞的增殖有關(guān)。 2. IA在腦缺血再灌注后早期對Notch信號通路各分子Notch1、NICD、PS1、HES1的表達具有促進作用，提示腦缺血再灌注后早期IA促進Notch信號通路的激活。
[Abstract]:Aim: to study the effect of Ilexonin A (Ilexonin A) on the regulation of Notch signal and the proliferation and differentiation of neural stem cells (NSCs) after cerebral ischemia-reperfusion in rats, and to explore the repair mechanism after brain injury. Methods: the middle cerebral artery occlusion of left middle cerebral artery (MCAO) was established for 2 h. SD rats were randomly divided into normal group, sham operation group, model group and IA group. The model group (IA group) was perfused with 40 mg / kg IA for different time after reperfusion. The neurological function score was used to evaluate the recovery of neural function in rats. TTC staining was used to verify the experimental animal model and to provide localization markers. The number of positive cells co-expressed by Brdu, Nestin, a neural stem cell marker, and the number of positive cells co-expressed in neuronal marker neuronal nuclear nucleus nestin by immunofluorescence assay and Notch signal transduction. The number of positive cells of Notch intracellular domain (NICD) in the intracellular domain of pathway molecule Notch signal, Western blot was used to detect the expression of NICD and RT-PCR was used to detect the expression of Notch1 receptor protein, presenilin PS1 protein and target gene HES1 in Notch signaling pathway. Results (1) recovery of neural function: the neurological deficit score of model group and IA group increased significantly after 1 day of cerebral ischemia reperfusion, and decreased gradually with the increase of time. Compared with model group on the 3rd day after reperfusion, the proliferation of Brdu/Nestin positive cells in ischemic peripheral area was significantly increased in group IA and model group at day 7 and day 7. The expression of Brdu/Nestin positive cells in ischemic peripheral area of group A was significantly higher than that in group A at 7 days after reperfusion (P < 0.05), and that in the model group was significantly lower than that in group A on the 3rd day after reperfusion (P < 0.05), and there was no significant difference in the expression of Brdu/Nestin positive cells between the control group and the model group. On the 14th day after reperfusion, the Brdu/Neun positive cells in the IA group and the model group had only a small amount of expression and no significant difference. The activation of Notch signaling pathway and the expression of the genes in the model group and IA group were compared with those in the normal group and the sham operation group. The expression of Notch signal pathway molecules Notch1, PS1 and HES1 in the model group was higher than that in the normal group and sham operation group. The expression of Notch signal pathway molecules in the model group was higher than that in the control group and sham operation group. Conclusion. 1. IA promoted the recovery of neural function in rats after cerebral ischemia-reperfusion, and its mechanism was related to the proliferation of Nestin positive cells in ischemic peripheral area. 2. IA promoted the expression of Notch1NICD1PS1HES1 in the Notch signal pathway early after cerebral ischemia-reperfusion, suggesting that IA promoted the activation of Notch signal pathway in the early stage of cerebral ischemia-reperfusion.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R743.3


本文編號：1609468