氧化应激与内质网应激在小鼠心肌功能中的研究

发布时间：2018-01-04 18:42

本文关键词：氧化应激与内质网应激在小鼠心肌功能中的研究　出处：《河北大学》2009年硕士论文　论文类型：学位论文

【摘要】：氧化应激和内质网应激已经被牵涉到心脏病领域,尽管二者之间的关系仍然不很明确。本实验采用野生型Friend virus B (FVB)小鼠和金属硫蛋白(MT)转基因小鼠来研究谷胱甘肽缺失引起的氧化应激对内质网应激和心肌收缩功能的影响。FVB和MT转基因小鼠分别用GSH合成酶抑制剂丁硫堇(Buthionine sulfoximine, BSO)食用水溶液(30mmol/L)喂养两周。分别通过对GSH/GSSG、活性氧自由基(reactive oxygen species, ROS)、caspase-3活性的检测以及蛋白免疫印记、Langendorff心脏离体灌流(LVDP和±dP／dt指标)以及电镜技术来对氧化应激、细胞凋亡、内质网应激、心肌功能和心室超微结构进行评定。结果发现BSO显著降低了GSH/GSSG比率,增加了ROS的产生,从而巩固了氧化应激。心肌功能和心室超微结构在BSO处理后也发生了显著的变化,而这些损伤明显被MT所减轻。BSO通过上调BiP、calreguln、p-IRE1α和p-eIF2α蛋白的表达而触发了内质网应激,但是并没有影响IRE1α和eIF2α总蛋白的表达量。同时BSO引起了CHOP/GADD153、caspase-12、Bax蛋白表达量的上调并增加了caspase-3的活性,减少了Bcl-2和p-JNK蛋白的表达,从而引起细胞凋亡,而MT明显改善了细胞凋亡的程度。另外,抗氧化剂N-乙酰半胱氨酸(N-acetylcysteine, NAC)和内质网应激抑制剂牛熊去氧胆酸(tauroursodeoxycholic acid, TUDCA)明显扭转了氧化应激诱导剂甲萘醌(menadione)引起的心肌收缩功能降低的趋势。综上所述,所有这些数据表明内质网应激在引发心肌功能损伤中可能位于氧化应激下游起作用。
[Abstract]:Oxidative stress and endoplasmic reticulum stress have been implicated in cardiovascular diseases, although the relationship between the two is not clear. In this experiment, the wild type Friend virus B (FVB) mice and metallothionein (MT) oxidative stress in mice transgenic mice to study the effect of glutathione depletion of endoplasmic reticulum stress and myocardial contraction the function of.FVB and MT respectively with GSH synthase inhibitor buthionine sulfoximine (Buthionine sulfoximine, BSO) in drinking water (30mmol/L) for two weeks respectively. Based on GSH/GSSG, active oxygen free radicals (reactive oxygen, species, ROS), to detect the activity of Caspase-3 and Western blot, Langendorff isolated heart perfusion (LVDP and + dP / dt index) and electron microscopy of oxidative stress, apoptosis, endoplasmic reticulum stress, myocardial function and ventricular ultrastructure were evaluated. The results show that BSO significantly reduced GSH/GSSG The ratio increased ROS production, thereby consolidating oxidative stress. Myocardial function and ventricular ultrastructure also changed significantly after BSO treatment, and the obvious injury by MT reduce.BSO through upregulation of BiP, calreguln, p-IRE1 and p-eIF2 expression of alpha alpha protein triggered endoplasmic reticulum stress, but did not the expression of IRE1 and alpha eIF2 alpha total protein amount. At the same time, caused by BSO CHOP/GADD153, caspase-12, Bax protein expression was up-regulated and increased the activity of Caspase-3, reduce the expression of Bcl-2 and p-JNK proteins, which induce cell apoptosis, and MT significantly improved the extent of apoptosis. In addition, the antioxidant N-acetylcysteine (N- N-acetylcysteine, NAC) and endoplasmic reticulum stress inhibitor tauroursodeoxycholic acid (tauroursodeoxycholic acid, TUDCA) significantly reversed the oxidative stress inducer (menadione) induced by myocardial systolic function A downward trend. In summary, all of these data suggest that endoplasmic reticulum stress may play a role in the cause of myocardial dysfunction that may be downstream of oxidative stress.

【学位授予单位】：河北大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R363

【引证文献】