富氢生理盐水对大鼠蛛网膜下腔出血后早期脑损伤的神经保护作用及机制研究

发布时间：2018-03-08 01:35

本文选题：蛛网膜下腔出血　切入点：早期脑损伤　出处：《浙江大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的：早期脑损伤(Early brain injury, EBI)在蛛网膜下腔出血(subarachnoid hemorrhage, SAH)发病机制中扮演着重要作用。神经元凋亡是参与SAH后EB]的重要病理机制。前期研究表明,富氢生理盐水(Hydrogen-rich Saline, HS)能抑制神经元凋亡,从而减轻SAH后的EBI。氢气在EBI中具体的抗凋亡效应的分子机制至今仍未被阐明。在本研究中,我们旨在阐述氢气是否能通过调节Akt/GSK3β信号通路的活性来抑制神经元凋亡,进而减轻SAH后的EBI。方法： Sprague-Dawley大鼠(n=85)被随机分成以下几组：假手术组(n=17), SAH组(n=17), SAH+生理盐水组(n=17),SAH+HS组(n=17)和SAH+HS+Ly294002组(n=17)。HS或者等量的生理盐水立即被注射到术后的大鼠体内,8小时后重复注射一次。P13K抑制剂Ly294002用来抑制上述的信号通路。SAH后24小时进行死亡率计算、神经功能评分及SAH分级。应用Western blot量化分析Akt、pAkt、GSK3β、pGSK3β和caspase-3等蛋白。通过末端脱氧核糖核酸染色法(TUNEL)和神经元特异性核蛋白(NeuN)来检测神经元凋亡。用免疫组化来阐明神经元凋亡和pAkt或者pGSK3β之间的关系。结果： HS能显著地减少神经元凋亡,从而改善SAH大鼠的神经功能缺失。HS能使在神经元表达的pAkt和pGSK3β水平显著地上调了。除此之外,通过HS的治疗,caspase-3显著下调。通过pAkt和TUNEL的染色显示了pAkt阳性细胞和TUNEL阳性细胞的共同表达。P13K抑制剂Ly294002能够抑制HS带来的良好效应。结论： HS通过激活Akt/GSK3β通路,能够减轻EBI的神经元凋亡,改善SAH后的神经功能预后。
[Abstract]:Objective:. Early brain injuryplays an important role in the pathogenesis of subarachnoid hemorrhage (Sah). Neuronal apoptosis is an important pathological mechanism involved in EB after SAH. Hydrogen-rich Salineine (HS) can inhibit neuronal apoptosis and alleviate EBI. the molecular mechanism of anti-apoptotic effect of hydrogen in EBI has not been elucidated. We aim to investigate whether hydrogen can inhibit neuronal apoptosis by regulating the activity of Akt/GSK3 尾 signaling pathway, and then reduce the EBI after SAH. Methods:. Sprague-Dawley rats were randomly divided into the following groups: sham-operated group (n = 17) and SAH HS Ly294002 group (n = 17) and SAH HS Ly294002 group (n = 17) and SAH HS Ly294002 group (n = 8 h after injection). P13K inhibitor Ly294002 was used to inhibit the above signaling pathway. SAH was used to calculate the mortality rate 24 hours later. Nerve function score and SAH grade. Western blot was used to quantitatively analyze the proteins, such as Aktaphane, GSK3 尾, pGSK3 尾 and caspase-3. Apoptosis of neurons was detected by terminal deoxyribonucleic acid staining (Tunel) and neuron specific nucleoprotein (Neun). The neuronal apoptosis was elucidated by immunohistochemistry. The relationship between meta apoptosis and pAkt or pGSK3 尾. Results:. HS can significantly reduce neuronal apoptosis, thereby improving neural function loss in SAH rats. HS can significantly up-regulate the expression of pAkt and pGSK3 尾 in neurons. Through the treatment of HS, caspase-3 was significantly down-regulated. PAkt and TUNEL staining showed that the co-expression of pAkt positive cells and TUNEL positive cells. P13K inhibitor Ly294002 could inhibit the good effect of HS. Conclusion:. HS can attenuate the neuronal apoptosis of EBI by activating Akt/GSK3 尾 pathway and improve the prognosis of neural function after SAH.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.15

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