發(fā)布時間：2018-04-25 06:22

  本文選題：熒光分析法 + 羥自由基��； 參考：《山東農業(yè)大學》2009年碩士論文

【摘要】： 生命活動的代謝過程中不斷產生各種活性氧自由基(ROS),羥基自由基(·OH),超氧自由基(O_2·~-)是其中具有代表性的兩種自由基,它們與生物體的衰老及許多疾病的發(fā)生都有重要關系。 ·OH是一種氧化能力很強的自由基,能很容易地氧化各種有機物和無機物,氧化效率高,反應速率快,是造成組織脂質過氧化、核酸斷裂、蛋白質和多糖分解的活性氧,與機體的衰老、腫瘤、輻射損傷和細胞吞噬有關。O_2·~-不僅自身具有毒性,而且可以經過一系列反應生成其它活性氧自由基,進一步對生物體產生損傷作用,且停留時間較長。 人體內總自由基中約95%以上屬于活性氧自由基,因此活性氧自由基的研究對人體有著特殊的意義。通過對活性氧自由基的檢測研究,可以幫助我們更好的解釋疾病產生的原因,加強有效的預防措施,減少機體的傷害程度。另外在衡量保健食品的抗氧化能力和中草藥抗氧化物質的篩選中具有重要的指導意義和理論價值。 活性氧自由基反應性強,存在時間短,不易進行直接測定。目前國內外已有較多間接測定·OH和O_2·~-的方法。本研究在全面了解這些方法的基礎上,研究了利用熒光分析技術測定活性氧自由基的方法,因為此技術具有靈敏度高、選擇性好、操作簡便等優(yōu)點。本文合成了兩種新型熒光探針,同時對這兩種探針進行了檢測選擇性的研究,探討了部分抗氧化劑對活性氧自由基的清除作用。具體研究內容分為以下三點: (1)通過鳥苷與過氧化氫光解反應,形成熒光物質8-羥基鳥苷,分析8-羥基鳥苷在不同條件下存在的形式及不同存在形式與熒光強度的關系。探討羥基自由基產生及其與鳥苷反應的條件,根據熒光強度與羥基自由基的量效關系,研究羥基自由基與鳥苷作用的分子機理,探討茶葉提取液和大蒜提取液通過清除羥基自由基對鳥苷的保護作用。 (2)設計合成一種新的熒光探針試劑—2-(2′-吡啶亞胺甲基)苯酚,建立檢測O_2·~-和SOD活性的熒光方法,研究測定方法的最佳條件,并用于大蒜、菠菜、木瓜中SOD活性的測定,并與鄰苯三酚自氧化法測定出的SOD活性進行了比較。 (3)建立了一種以2-(2′-噻吩基)苯并噻唑啉為新型熒光探針測定O_2·~-和SOD活性的熒光分析方法,探討了其它物質對體系的干擾影響及熒光探針的檢測單一性,研究測定方法的靈敏度,精密度,本法成功應用于大蒜、洋蔥等生物樣品中SOD活性的測定。 本論文首次對羥基自由基與鳥苷作用分子機理的熒光分析做了相關研究,并合成了新型熒光探針,用于O_2·~-和SOD活性的測定,且2-(2′-噻吩基)苯并噻唑啉對O_2·~-的檢測具有專一性,得到較為滿意的結果,但是也存在一些不足之處,如鳥苷與過氧化氫光解反應體系中測定?OH的含量時靈敏度還不夠高,還需進一步探討�；钚匝踝杂苫臏y定方法還未能很好的應用于動物體的檢測,有待進一步完善。
[Abstract]:Active oxygen free radicals (ROS), hydroxyl radical (/ OH) and superoxide radical (O_2 - -) are two free radicals in the metabolic process of life activities. They are closely related to the aging of organisms and the occurrence of many diseases.
OH is a highly oxidizing free radical, which can easily oxidize various organic and inorganic substances. The oxidation efficiency is high and the reaction rate is fast. It is the active oxygen that causes tissue lipid peroxidation, nucleic acid breakage, protein and polysaccharide decomposition. It is related to body aging, tumor, radiation injury and cell phagocytosis. Not only is it related to its own toxicity, but also related to body aging, tumor, radiation injury and cell phagocytosis. Moreover, a series of reactions can produce other reactive oxygen radicals, which can further damage the organism and stay longer.
More than 95% of the total free radicals in the human body belong to the reactive oxygen free radicals, so the study of reactive oxygen radicals is of special significance to the human body. Through the study of the detection of reactive oxygen radicals, we can help us to better explain the causes of the disease, strengthen effective prevention measures, reduce the injury degree of the body. In addition, it is also a measure of the protection of the body. The antioxidant capacity of healthy foods and the screening of antioxidants in Chinese herbal medicines are of great guiding significance and theoretical value.
Active oxygen radicals have strong reactivity, short existence time and not easy to direct determination. At present, there are many methods of indirect determination of OH and O_2. In this study, based on the comprehensive understanding of these methods, the method for the determination of reactive oxygen radicals by fluorescence analysis technology is studied, because this technique has high sensitivity and good selectivity. Two new fluorescent probes were synthesized in this paper. At the same time, the detection selectivity of these two probes was studied and the scavenging effect of some antioxidants on reactive oxygen radicals was discussed. The specific research contents were divided into three points as follows:
(1) 8- hydroxyguanosine was formed by the photodissociation of guanosine and hydrogen peroxide, and the relationship between the form of 8- hydroxy guanosine under different conditions and the relationship between different forms and fluorescence intensity was analyzed. The production of hydroxyl radicals and the conditions for the reaction with guanosine were discussed. According to the quantitative relationship between the fluorescence intensity and the hydroxyl radical, the hydroxyl group was studied. The molecular mechanism of free radical interaction with guanosine was studied. The protective effects of tea extract and garlic extract on hydroxyl radical scavenging activity of guanosine were investigated.
(2) to design and synthesize a new fluorescent probe reagent, 2- (2 '- pyridimimomethyl) phenol, to establish a fluorescence method for detecting the activity of O_2 - and SOD, to study the optimum conditions for the determination of the method and to determine the activity of SOD in garlic, spinach and papaya, and to compare the activity of SOD with the self oxidation of phthalic three.
(3) a fluorescence analysis method was established by using 2- (2 '- thiophenyl) benzothiazoline as a new fluorescent probe for the determination of O_2. - and SOD activity. The interference effects of other substances on the system and the singularity of the detection of fluorescence probe were discussed. The sensitivity and precision of the method were studied. This method was successfully applied to the biological samples of garlic and onion, such as SOD Determination of activity.
In this paper, the fluorescence analysis of the molecular mechanism of hydroxyl radical and guanosine was studied for the first time. A new fluorescent probe was synthesized and used to determine the activity of O_2 - and SOD. The detection of 2- (2 '- thiophenyl) benzothiazoline has a specificity and satisfactory results for the detection of O_2 - - but there are some shortcomings, such as birds. The sensitivity of the determination of OH in the photodissociation reaction system of glucoside and hydrogen peroxide is not high enough, and further discussion is needed. The determination method of reactive oxygen free radicals is not well applied to the detection of animal body, which needs further improvement.

【學位授予單位】：山東農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2009
【分類號】：R341

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本文編號：1800116