羰基应激与红细胞衰老机制研究

发布时间：2018-01-02 01:10

本文关键词：羰基应激与红细胞衰老机制研究　出处：《湖南师范大学》2010年硕士论文　论文类型：学位论文

【摘要】： 脂质过氧化和非酶糖基化作为体内能量代谢过程中两大必然发生的生化副反应,会产生一系列的活性羰基类物质。这些活性羰基类物质具有很高的生物活性,很容易与生物大分子的巯基和氨基发生反应,导致生物大分子的交联。在血液中,这些活性羰基类物质能攻击血液中的许多生物大分子,如攻击红细胞膜蛋白,导致红细胞的结构和功能发生改变。因而,有研究者认为：羰基应激造成的生物大分子的失修性损伤是导致机体衰老的重要原因。本学位论文研究工作分别从红细胞体外快速衰老(新鲜血液体外储存)和红细胞体内衰老(不同年龄人群)两个方面着手,探讨羰基应激与红细胞衰老的关系。与衰老过程中血液流变学改变类似,体外储存血液的粘度会随着储存时间的延长逐渐上升,然而其具体生化机制仍不明了。为了探索这种粘度上升的机制,本文检测了血液在储存过程中活性羰基类物质含量、硫醇含量、红细胞膜蛋白羰基化程度以及蛋白质荧光强度的变化。结果显示：随着血液体外储存时间的延长,活性羰基类物质含量与膜蛋白羰基化程度逐渐增加,而硫醇水平与蛋白质荧光强度逐渐降低。这些结果表明：随着红细胞体外储存时间的延长,活性羰基类物质的含量上升,它们很容易与血液中的生物大分子特别是红细胞膜蛋白的巯基和氨基发生反应,引起蛋白交联,进而使红细胞的结构改变,导致血液粘度增加。为了验证体外试验的结果,并进一步研究红细胞衰老机制,本文对不同年龄人群新鲜血液进行了一系列检测分析。首先对整体样本血液中活性羰基类物质、还原性硫醇以及部分样本红细胞膜蛋白羰基化程度进行测定,结果与体外试验一致：随着年龄的增长,血液中活性羰基类物质含量和红细胞膜蛋白羰基化程度增加,而还原性硫醇水平逐渐下降。其次本文分析了同一人群的平均红细胞血红蛋白浓度、平均红细胞血红蛋白含量、血浆中主要的脂类代谢相关物质与人红细胞膜蛋白羰基化程度的关系。结果显示：红细胞膜蛋白羰基化程度与平均红细胞血红蛋白浓度及含量基本没有相关性,与高密度脂蛋白存在一定负相关,而与甘油三酯含量、总胆固醇含量、总胆固醇/HDL均存在一定的正相关。这些结果表明：人红细胞膜蛋白羰基化程度与血液中脂类物质的含量正相关而与红细胞的蛋白质含量无关,这间接证明了红细胞膜蛋白羰基化与脂质过氧化生成的活性羰基类物质相关。以上的结果表明：血液中活性羰基类物质水平会随年龄的增加而增加,由于其具有很强的生物反应活性,很容易攻击红细胞膜蛋白的氨基和巯基,引起蛋白交联,导致红细胞结构及生物功能的改变。这很可能是红细胞衰老的重要机制之一。
[Abstract]:Lipid peroxidation and non-enzymatic glycosylation are two inevitable biochemical side reactions in the process of energy metabolism in vivo, resulting in a series of active carbonyl substances, which have high biological activity. It is easy to react with the sulfhydryl and amino groups of biomolecules, leading to crosslinking of biomolecules. In the blood, these active carbonyl compounds can attack many biological macromolecules in the blood, such as attacking erythrocyte membrane proteins. As a result, some researchers believe that the damage of biomolecules caused by carbonyl stress is an important cause of aging. The research work of this dissertation is from two aspects: rapid aging of red blood cells in vitro (fresh blood in vitro storage) and in vivo aging of red blood cells (people of different ages). To explore the relationship between carbonyl stress and erythrocyte senescence. Similar to the changes in hemorheology during aging, the viscosity of blood stored in vitro will increase with the prolongation of storage time, but its specific biochemical mechanism is still unclear. In this paper, the contents of active carbonyl substances and mercaptan in blood during storage were determined. The changes of erythrocyte membrane protein carbonylation degree and protein fluorescence intensity. The results showed that the content of active carbonyl substances and the degree of membrane protein carbonylation increased with the prolongation of blood storage time in vitro. The level of mercaptan and the fluorescence intensity of protein decreased gradually. These results showed that the content of active carbonyl compounds increased with the prolongation of the storage time of erythrocytes in vitro. They react easily with the mercapto and amino groups of the biological macromolecules in the blood, especially the sulfhydryl and amino groups of the erythrocyte membrane protein, which leads to the crosslinking of the proteins, which changes the structure of the red blood cells and results in the increase of the blood viscosity. In order to verify the results of in vitro tests and further study the mechanism of erythrocyte aging, a series of tests were carried out on the fresh blood of different age groups. Firstly, the active carbonyl substances in the whole blood samples were analyzed. The degree of carbonylation of reductive mercaptan and some samples of erythrocyte membrane protein was determined and the results were consistent with the in vitro test: with the increase of age. The content of active carbonyl substances in blood and the degree of carbonylation of erythrocyte membrane protein increased, while the level of reductive mercaptan decreased gradually. Secondly, the mean erythrocyte hemoglobin concentration of the same population was analyzed. Mean erythrocyte hemoglobin content. The results showed that there was no correlation between the level of erythrocyte membrane protein carbonylation and the average erythrocyte hemoglobin concentration and content. There was a negative correlation with high density lipoprotein, but with triglyceride content and total cholesterol content. The results showed that the degree of carbonylation of human erythrocyte membrane protein was positively correlated with the content of lipids in blood, but not with the protein content of erythrocyte. These results suggest that the carbonylation of erythrocyte membrane proteins is related to the active carbonyl substances produced by lipid peroxidation. The above results indicate that the level of active carbonyl substances in blood increases with age. Because of its strong biological reaction activity, it is easy to attack the amino and sulfhydryl groups of erythrocyte membrane protein, leading to protein cross-linking. This may be one of the important mechanisms of erythrocyte aging.
【学位授予单位】：湖南师范大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R33

【相似文献】