Non-Invasive Probing Cellular Oxidative Stress and Antioxidants Therapeutic Effectiveness

非侵入性探测细胞氧化应激和抗氧化剂的治疗效果

• 批准号: 10652764
• 负责人: Anhong Zhou
• 金额: $ 41.47万
• 依托单位: UTAH STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-15 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10652764
• 关键词: Acceleration; Air Pollutants; Air Pollution; Animals; Antioxidants; Biochemical; Biological; Biological Assay; Biological Markers; Biological Models; Biomedical Engineering; Biomedical Research; Carbohydrates; Carcinogens; Cardiovascular Diseases; Cells; Collaborations; Cytoprotection; DNA; Data; Development; Diesel Exhaust; Dietary Intervention; Dietetics; Disease; Docosahexaenoic Acids; Engine Exhaust; Epithelial Cells; Exposure to; Fingerprint; Fluorescence; Fluorescent Dyes; Generations; Goals; Health; Human; In Situ; In Vitro; Inflammatory; Inhalation; Inhalation Exposure; Injury; International Agency for Research on Cancer; Knowledge; Label; Learning; Link; Lipid Peroxidation; Lipids; Lung; Lung diseases; Machine Learning; Malignant Neoplasms; Malignant neoplasm of lung; Mathematics; Measurement; Measures; Mediating; Methodology; Methods; Molecular; Mus; Nucleic Acids; Oxidative Stress; Oxidative Stress Induction; Particulate; Particulate Matter; Peer Review; Photobleaching; Polyunsaturated Fatty Acids; Process; Production; Proteins; Public Health; Publications; Pulmonary Heart Disease; Raman Spectrum Analysis; Reactive Oxygen Species; Research; Research Personnel; Research Project Grants; Resolution; Respiratory System; Resveratrol; Role; Specificity; Structure of parenchyma of lung; Students; System; Techniques; Technology; Testing; Therapeutic; Therapeutic Agents; Tissues; Toxic effect; Toxicology; Validation; Work; World Health Organization; biological adaptation to stress; cancer classification; carcinogenicity; cell injury; cell type; effectiveness evaluation; experimental study; food science; in vivo; machine learning algorithm; multidisciplinary; nanoprobe; new technology; novel therapeutics; nutrition; oxidative damage; particle; particle exposure; peroxidation; programs; respiratory; response; screening; skills training; statistics; symposium; therapeutic effectiveness; training opportunity; undergraduate student

PROJECT SUMMARY. Particulate matter (PMs) that induces oxidative stress have prominent impacts and adverse health effects including increased cardiovascular and pulmonary diseases. The World Health Organization recognizes diesel engine particles (DEPs) as mutagenic and carcinogenic to humans and the International Agency for Research on Cancer (IARC) classified diesel engine exhaust as a human group I carcinogen. It has been established that generation of reactive oxygen species (ROS) and oxidative stress is an important toxicological mechanism of particulate air pollution-induced lung cancer and disorders. However, the pathophysiological mechanism of air pollutant mediated pulmonary toxicity still remains unclear. This is primarily due to the lack of efficient test systems, mimicking human air pollutant inhalation exposure scenarios. Furthermore, it is unclear, but important, to understand whether epithelial cells from different respiratory tract regions respond to PMs similarly or differently and how to develop measurement technology to assess responses in both in vitro and in vivo. Dietary interventions with antioxidants (e.g., resveratrol), polyunsaturated fatty acids (PUFA, such as docosahexaenoic acid or DHA) are an attractive therapeutic approach to ameliorate air pollution-induced respiratory toxicity. Current methods to detect cellular ROS are fluorescence-based, suffering several drawbacks: each fluorescent dye is limited to a couple of ROS species, not detecting the overall ROS production; single-cell resolution is not achievable; lack of both molecule-level details and temporal specificity; and the photobleaching problem. The primary goal of this proposal is to test the application of a non-invasive, non-destructive, label-free Raman spectroscopy technique in combination with machine learning analysis to assess cellular oxidative stress and cytoprotection by antioxidants. Our central hypothesis is that subcellular and biochemical composition alterations induced by oxidative stress and intracellular ROS production can be quantitatively detected in the cell by Raman spectroscopy using the combinational analysis of respective spectral markers of the fingerprint and the silent regions. To accomplish this goal, we propose the following specific aims: 1) In vitro toxicity assessment of epithelial cells exposed to DEP and the effects of antioxidants; 2) Identification of oxidative stress-associated spectral markers in the fingerprint region and development of intracellular ROS sensing nanoprobes in the silent region; and 3) Pre-validation of Raman-derived spectral markers and ROS quantitation on the lung tissues isolated from DEP exposed mice. We expect that successful completion of these aims will yield a wealth of new knowledge regarding the mechanistic link(s) between cellular oxidative stress biomarkers and non-physiological spectral markers. Further, we expect that these experiments will establish a new technical platform for screening antioxidant activities that can benefit the discovery of new therapeutic agents to battle PM-induced lung disease and other oxidative stress induced inflammatory diseases.

项目摘要。 引起氧化应激的颗粒物 (PM) 具有显着的影响和不利的健康影响 包括增加心血管和肺部疾病。世界卫生组织承认柴油 发动机颗粒（DEP）对人类和国际研究机构具有致突变性和致癌性 癌症研究中心 (IARC) 将柴油发动机废气列为人类 I 类致癌物。已经确定 活性氧（ROS）和氧化应激的产生是重要的毒理学机制 颗粒空气污染诱发肺癌和疾病。然而，空气的病理生理机制 污染物介导的肺毒性仍不清楚。这主要是由于缺乏有效的测试 系统，模仿人类空气污染物吸入暴露场景。此外，尚不清楚但很重要的是， 了解不同呼吸道区域的上皮细胞对 PM 的反应是否相似或 不同的方式以及如何开发测量技术来评估体外和体内的反应。饮食 使用抗氧化剂（例如白藜芦醇）、多不饱和脂肪酸（PUFA，例如二十二碳六烯酸）进行干预 酸或 DHA）是改善空气污染引起的呼吸道毒性的一种有吸引力的治疗方法。 目前检测细胞 ROS 的方法是基于荧光的，有几个缺点： 染料仅限于几种 ROS 种类，无法检测总体 ROS 产生；单细胞分辨率不 可以实现的；缺乏分子水平的细节和时间特异性；和光漂白问题。这 该提案的主要目标是测试非侵入性、非破坏性、无标记拉曼的应用 光谱技术与机器学习分析相结合，评估细胞氧化应激和 抗氧化剂的细胞保护作用。我们的中心假设是亚细胞和生化成分的改变 拉曼可以定量检测细胞内氧化应激诱导的细胞内ROS产生 使用指纹和静音各自光谱标记的组合分析进行光谱分析 地区。为了实现这一目标，我们提出以下具体目标：1）体外毒性评估 暴露于 DEP 的上皮细胞和抗氧化剂的作用； 2) 氧化应激相关的鉴定 指纹区域的光谱标记和无声环境下细胞内ROS传感纳米探针的开发 地区; 3) 肺组织上拉曼衍生光谱标记和 ROS 定量的预验证 从暴露于 DEP 的小鼠中分离得到。我们预计这些目标的成功实现将产生大量新的成果 关于细胞氧化应激生物标志物与非生理学之间的机制联系的知识 光谱标记。此外，我们预计这些实验将建立一个新的筛选技术平台 抗氧化活性有助于发现新的治疗药物来对抗 PM 引起的肺部疾病 和其他氧化应激诱发的炎症性疾病。