【摘要】： 衰老是遺傳因素和環(huán)境因素共同作用的結(jié)果。根據(jù)羰基應(yīng)激衰老學(xué)說(shuō)的觀點(diǎn),在氧自由基引起的脂質(zhì)過(guò)氧化和非酶糖基化反應(yīng)過(guò)程中,會(huì)生成具有反應(yīng)活性的共同中間產(chǎn)物——活性羰基類物質(zhì),它們能與生物體內(nèi)的幾乎所有生物大分子發(fā)生羰-氨交聯(lián),這一過(guò)程是機(jī)體內(nèi)衰老損變的核心過(guò)程,甚至是機(jī)體衰老改變的生化本質(zhì)。 本學(xué)位論文以分別代表兩個(gè)極端的正常細(xì)胞(即體外生存能力很弱的紅細(xì)胞和長(zhǎng)期生活在體外的酵母)為實(shí)驗(yàn)?zāi)Ｐ?探討它們對(duì)羰基應(yīng)激的響應(yīng)差異。 一、血液在儲(chǔ)存過(guò)程中,血液粘度會(huì)隨著儲(chǔ)存時(shí)間的延長(zhǎng)逐漸上升,然而其具體生化機(jī)制仍不明了。為了探索這種血液粘度升高的機(jī)制,本文探索了血液儲(chǔ)存過(guò)程中硫醇、TBARS水平、紅細(xì)胞膜蛋白羰基化程度以及蛋白質(zhì)熒光強(qiáng)度的變化。結(jié)果顯示:伴隨著血液粘度的升高,硫醇水平與蛋白質(zhì)熒光強(qiáng)度逐漸降低,而TBARS水平與膜蛋白羰基化程度逐漸增加。這些結(jié)果表明:活性羰基類物質(zhì)很容易攻擊蛋白質(zhì)的氨基和巰基,羰基應(yīng)激誘導(dǎo)的紅細(xì)胞膜蛋白羰基化可能是血液儲(chǔ)存過(guò)程中粘度上升的重要機(jī)制。二、為了探索酵母細(xì)胞對(duì)羰基應(yīng)激的響應(yīng),本文用不同濃度的MDA溶液處理酵母細(xì)胞,培養(yǎng)過(guò)夜后用紫外分光光度計(jì)在600 nm處測(cè)定培養(yǎng)過(guò)夜后酵母的光密度,同時(shí)用PI熒光染色法(Ex530/Em590)測(cè)定經(jīng)MDA處理后的酵母細(xì)胞的活力。結(jié)果顯示:即使在高濃度MDA的作用下,酵母細(xì)胞生長(zhǎng)和活力均未受到顯著抑制。這表明:酵母細(xì)胞抗羰基應(yīng)激的能力很強(qiáng),但這種抗性的具體機(jī)制仍需做進(jìn)一步的實(shí)驗(yàn)探討。 綜上所述,血液中的紅細(xì)胞在羰基應(yīng)激條件下會(huì)發(fā)生一系列顯著的生理生化改變;而生長(zhǎng)環(huán)境相對(duì)較惡劣的酵母卻對(duì)羰基應(yīng)激不敏感。
[Abstract]:Aging is the result of both genetic and environmental factors. According to the theory of carbonyl stress senescence, in the process of lipid peroxidation and non-enzymatic glycosylation induced by oxygen free radicals, a common intermediate product with reactive activity, active carbonyl compounds, is produced. They can be crosslinked with almost all biological macromolecules in organism. This process is the core process of aging in organism and even the biochemical essence of aging change in organism. In this dissertation, two extreme normal cells (red blood cells with weak viability in vitro and yeast living in vitro for a long time) were used as experimental models to explore their responses to carbonyl stress. One is that the viscosity of blood increases gradually with the prolongation of storage time, but the specific biochemical mechanism is still unclear. In order to explore the mechanism of the increase of blood viscosity, the changes of TBARS level, carbonylation degree of erythrocyte membrane protein and the fluorescence intensity of protein during blood storage were investigated. The results showed that with the increase of blood viscosity, mercaptan level and protein fluorescence intensity decreased gradually, while TBARS level and carbonylation degree of membrane protein increased gradually. These results suggest that active carbonyl compounds can easily attack the amino and mercapto groups of proteins. Carbonylation of erythrocyte membrane proteins induced by carbonyl stress may be an important mechanism of viscosity increase during blood storage. Secondly, in order to explore the response of yeast cells to carbonyl stress, yeast cells were treated with different concentrations of MDA solution. After overnight cultivation, the light density of yeast was measured by UV spectrophotometer at 600 nm. At the same time, the activity of yeast cells treated with MDA was determined by PI fluorescence staining (Ex530/Em590). The results showed that the growth and activity of yeast cells were not significantly inhibited even under high concentration of MDA. This indicates that yeast cells have a strong ability to resist carbonyl stress, but the specific mechanism of this resistance needs to be further studied. To sum up, red blood cells in the blood will undergo a series of significant physiological and biochemical changes under carbonyl stress, while yeast, which has a relatively bad growth environment, is not sensitive to carbonyl stress.
【學(xué)位授予單位】：湖南師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R363

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本文編號(hào)：2227291