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New Fluorescent Probes Based on Antioxidant Peptides and Amino Acids for Detecting Reactive Oxygen Species in Living Cells
Author: WangZuoZuo
Tutor: XuKeHua；TangBo
School: Shandong Normal University
Course: Analytical Chemistry
Keywords: Antioxidant peptide Histidine Reactive oxygen species Singlet oxygen Fluorescent probe Confocal microscopic imaging
CLC: Q503
Type: Master's thesis
Year: 2010
Downloads: 152
Quote: 0
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Abstract
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Will continue to produce a variety of reactive oxygen radicals in biological metabolic processes. Superoxide anion (O2-.), hydrogen peroxide (H2O2), hydroxyl radical (. OH), singlet oxygen (1O2), lipid peroxide radicals (ROO.) and nitric oxide (NO) is life important representative of free radicals, in the course of their organism aging and many diseases have an important association. The appropriate level of reactive oxygen species in the organism is the basis of the body to maintain a good immune response, while its large presence is harmful. For this reason, the study of intracellular reactive oxygen species in recent years has caused the people more and more attention, detection of reactive oxygen species has evolved, in which the fluorescence method is simple, easy to operate, with high sensitivity and selectivity. Role of reactive oxygen radicals within the fluorescent probe molecules and living cells to generate a strong fluorescence substance, by means of confocal laser imaging technology can achieve active molecules to be tested in the intracellular visualization, and thus their life in vivo greater, and thus the detection of the \low to reduce the damage of light on the cells, so the near-infrared fluorescent probes used to detect intracellular reactive oxygen species has good prospects for development. Fluorescence properties of this thesis is to design highly selective, high-sensitivity detection of intracellular singlet oxygen (1O2) near-infrared fluorescent probes for the purpose of using the fluorescent probe 1O2 specific role change, to achieve its detection, and the synthetic probe successfully for biological living cells by confocal microscopy imaging. This paper carried out the research work of the following two aspects: (a) of singlet oxygen scavenging antioxidant amino acid-based design and synthesis of a cyanine parent structure containing histidine new near-infrared fluorescent probes. Probe itself weak fluorescence, 1O2 reaction fluorescence significantly enhanced excitation and emission wavelengths at 754 nm and 794 nm, respectively. The probe can be captured with high selectivity 1O2, coexistence with other reactive oxygen species (ROS) and a number of biological samples is often the presence of a reducing compound and a metal ion does not interfere. This method is detect 1O2 linear range is 5.0 × 10-8-1.2 × 10-6 M, the detection limit is 99 nM. Successfully achieved by using a laser scanning confocal microscope, the probe cells 1O2 fluorescence imaging indicates that the probe has a good film penetrating, it is possible to selectively capture the intracellular 1O2, and may be on the cell within the 1O2 generate a response, and performance in a good biological applicability. (B) Design and Synthesis of a new water-soluble sulfonic acid group cyanine precursor structure containing carnosine better near-infrared fluorescent probes, for selectively detecting vivo 1O2. Imidazolyl is in the probe with singlet oxygen to the reactive groups, cause changes in the fluorescence of the probe, which can achieve the quantitative detection of the singlet oxygen. The probe has a good solubility in water and good biocompatibility and thus can be used for fluorescent imaging of the singlet oxygen in living cells. Currently, some experiments are still in progress.
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