The role of microRNA miR-21 in nickel nanoparticles-induced MMPs production

microRNA miR-21 在镍纳米粒子诱导的 MMP 产生中的作用

• 批准号: 8626878
• 负责人: Qunwei Zhang
• 金额: $ 45万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-17 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8626878
• 关键词: 17-(Allylamino)-17-demethoxygeldanamycin; Address; Adverse effects; Alloys; Area; Biological; Biosensor; Blood Cells; Blood Circulation; Bone Marrow; Breathing; Carbon; Cell Line; Cells; Data; Development; Diagnostic; Drug Formulations; Electrical battery; Electrodes; Engineering; Environmental Pollution; Exposure to; Gastrointestinal tract structure; Gelatin Zymography; Gelatinase A; Gelatinase B; Growth; Health; Heat-Shock Proteins 90; Hematopoietic stem cells; Home environment; Human; Hyperthermia; Hypoxia Inducible Factor; Immune response; In Vitro; Inflammatory; Inflammatory Response; Information Technology; Knock-out; Knowledge; Lead; Leukocytes; Liquid substance; Location; Lung; Lymphoma; Magnetic Resonance Imaging; Magnetism; Matrix Metalloproteinases; Measures; Medical; Messenger RNA; Metal exposure; Metals; MicroRNAs; Modeling; Molecular; Mus; Nanotechnology; Nickel; Particulate; Play; ProMMP-9; Production; Rattus; Reporting; Risk; Role; Silicon Dioxide; Site; Skin; Small Interfering RNA; Solar Energy; Surface Properties; System; Testing; Time; Tissue Inhibitor of Metalloproteinase-1; Toxic effect; Toxin; Transfection; Transition Elements; U937 Cells; Up-Regulation; Work; absorption; base; catalyst; chemical property; cytokine; improved; in vitro Assay; in vivo; inhibitor/antagonist; interest; knock-down; macrophage; meetings; monoblast; monocyte; nano; nanomaterials; nanoparticle; nanosized; nanotoxicology; new technology; novel; particle; peripheral blood; physical property; public health relevance; titanium dioxide; tool; transcription factor

With the development of nanotechnology, a large number of metal nanoparticles are being developed and produced with new formulations and surface properties to meet novel demands. For example, as an important transition metal, nickel nanoparticles (Nano-Ni) have wide ranging applications in the fields of batteries, electrical conductors, permanent magnets, magnetic fluids, magnetic recording media, solar energy absorption, fuel cell electrodes, and catalysts. In addition, due to structural and inherent chemical and physical properties, nickel alloy nanomaterials have received special interest in biomedical applications. As the use of nanomaterials continues to grow, the risk of environmental contamination by nanoparticles is increases. The term 'Nanotoxicology' has been used to put forward the concept that nanoparticles might represent a unique class of particulate toxins that differ from conventional pathogenic particles. In this proposal, we selected Nano-Ni as a model metal nanoparticles because of its wide industrial interest and biological and medical applications. Based on our preliminary data, we hypothesize that Nano-Ni will activate monocytes/macrophages, altering the expression and activity of matrix metalloproteinases (MMPs) through hypoxia inducible factor 1a (HIF-1a) and microRNA miR-21. It is possible that structurally- tailored Ni-containing nanoparticles such as nickel alloy nanoparticles, could have fewer or even no effects. This would be very important to know. This project will use both in vitro and in vivo systems to address the following objectives. (1). Determine the alteration of MMPs and TIMPs expression and activity in monocytes/macrophages exposed to Nano-Ni or nickel alloy nanoparticles in vitro and in vivo. We will examine whether exposure to Nano-Ni will alter MMP-2, MMP-9, TIMP-1 and TIMP-2 production in both U937 cells (human leukemic monocyte lymphoma cell line) and monocytes isolated from mice, by using quantitative real-time PCR, gelatin zymography or reverse zymography assay in vitro. We will also determine the alteration of MMPs and TIMPs expression and activity in monocytes isolated from Nano-Ni-exposed mice. We will investigate whether nickel alloy nanoparticles, Nano-Ni core and silica or carbon shell, and silica or carbon supported Nano-Ni will reduce the Nano-Ni-induced activation of MMPs and toxic effect in vitro and in vivo settings. (2). Examine HIF-1a accumulation in monocytes/macrophages with exposure to Nano-Ni and the role of HIF-1a on the Nano-Ni-induced alteration of MMPs expression and activity. We will measure the HIF-1a level in U937 cells with exposure to Nano-Ni. To investigate the role of HIF-1a in Nano-Ni- induced MMPs expression and activity, we will employ the following strategies: (1) use of HIF-1a inhibitors such as heat-shock protein 90 (Hsp90); (2) use of HIF-1a-specific siRNA to knock-down HIF-1a expression; and (3) use of HIF-1a knock-out cells. (3). Investigate whether Nano-Ni-induced alteration of MMPs expression and activity through up-regulation of microRNA miR-21 by activation of HIF-1a. We will first identify whether exposure to Nano-Ni will cause up-regulation of miR-21 by quantitative real-time PCR. We will then investigate whether miR-21 modulates MMPs expression and activity by activation HIF-1a by using: (1) pre-miR-21 transfection to increase the abundance of miR-21 in the cells; (2) anti-miR-21 transfection to lower the level of miR-21 in the cells; and (3) use HIF decoy to block the activity of endogenous HIF transcription factor in the cells.

随着纳米技术的发展，大量的金属纳米粒子被开发出来，并 采用新配方和表面性能生产，以满足新的需求。例如，作为一个重要的 过渡金属镍纳米颗粒(Nano-Ni)在电池领域有着广泛的应用， 电导体、永磁体、磁性液体、磁记录介质、太阳能 吸附、燃料电池电极和催化剂。此外，由于结构和固有的化学和物理 近年来，镍合金纳米材料在生物医学领域的应用引起了人们的极大兴趣。因为使用了 纳米材料继续增长的同时，纳米粒子污染环境的风险也在增加。这个 纳米毒理学一词被用来提出这样的概念：纳米颗粒可能代表一种独特的 一类不同于传统致病颗粒的颗粒毒素。在这份提案中，我们选择了 纳米镍作为一种模型金属纳米粒子，因其具有广泛的工业应用价值和生物医学价值而备受关注 申请。根据我们的初步数据，我们假设纳米镍将被激活 单核/巨噬细胞，改变基质金属蛋白酶的表达和活性 通过低氧诱导因子1a(HIF-1a)和microRNA miR-21。有可能在结构上- 定制的含镍纳米颗粒，如镍合金纳米颗粒，可能含有更少甚至没有 效果。知道这一点是非常重要的。该项目将使用体外和体内系统来 实现以下目标。(1)。检测MMPs和TIMPs的表达和活性变化 在体外和体内暴露于纳米镍或镍合金纳米颗粒的单核/巨噬细胞中。我们 我将研究暴露于纳米镍是否会改变两者的MMP2、MMP9、TIMP-1和TIMP-2的产生 U937细胞(人白血病单核细胞淋巴瘤细胞系)和小鼠单核细胞 体外实时荧光定量聚合酶链式反应、明胶酶谱分析或反向酶谱分析。我们还将确定 纳米镍暴露小鼠单核细胞MMPs和TIMPs表达及活性的变化 我们将调查镍合金纳米颗粒、纳米镍核和二氧化硅或碳壳，以及二氧化硅或 碳负载纳米镍可降低纳米镍诱导的MMPs的活化及体外毒性作用 VIVO设置。(2)。纳米镍对单核/巨噬细胞HIF-1a蓄积的影响 以及HIF-1a在纳米镍诱导的MMPs表达和活性改变中的作用。我们会 检测纳米镍对U937细胞HIF-1a表达的影响。探讨缺氧诱导因子-1a在纳米镍中的作用 诱导MMPs的表达和活性，我们将采用以下策略：(1)使用HIF-1a抑制剂 如热休克蛋白90(Hsp90)；(2)利用HIF-1a特异性siRNA下调HIF-1a的表达； (3)使用HIF-1a基因敲除细胞。(3)。纳米镍是否引起基质金属蛋白酶的改变 通过激活HIF-1a上调microRNA miR-21的表达和活性。我们将首先 通过实时定量聚合酶链式反应确定纳米镍暴露是否会导致miR-21表达上调。我们会 然后研究miR-21是否通过激活HIF-1a调节MMPs的表达和活性：(1) 预转染miR-21以增加细胞内miR-21的丰度；(2)反转miR-21以降低 细胞内miR-21的水平；(3)利用HIF圈套阻断内源性HIF转录的活性 细胞中的因素。

项目成果

Cobalt nanoparticles induce lung injury, DNA damage and mutations in mice.

• DOI: 10.1186/s12989-017-0219-z
• 发表时间: 2017-09-18
• 期刊: Particle and fibre toxicology
• 影响因子: 10
• 作者: Wan R;Mo Y;Zhang Z;Jiang M;Tang S;Zhang Q
• 通讯作者: Zhang Q

Analysis of Nanomaterial Toxicity by Western Blot.

通过蛋白质印迹分析纳米材料毒性。

• DOI: 10.1007/978-1-4939-8916-4_10
• 发表时间: 2019
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Long,Gao;Mo,Yiqun;Zhang,Qunwei;Jiang,Mizu
• 通讯作者: Jiang,Mizu

Determination of Phosphorylated Histone H2AX in Nanoparticle-Induced Genotoxic Studies.

纳米颗粒诱导的基因毒性研究中磷酸化组蛋白 H2AX 的测定。

• DOI: 10.1007/978-1-4939-8916-4_9
• 发表时间: 2019
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Wan,Rong;Mo,Yiqun;Tong,Ruirui;Gao,Meiqin;Zhang,Qunwei
• 通讯作者: Zhang,Qunwei

Endoplasmic reticulum stress and oxidative stress are involved in ZnO nanoparticle-induced hepatotoxicity.

• DOI: 10.1016/j.toxlet.2015.02.004
• 发表时间: 2015-04-02
• 期刊: TOXICOLOGY LETTERS
• 影响因子: 3.5
• 作者: Yang, Xia;Shao, Huali;Liu, Weirong;Gu, Weizhong;Shu, Xiaoli;Mo, Yiqun;Chen, Xuejun;Zhang, Qunwei;Jiang, Mizu
• 通讯作者: Jiang, Mizu

Application of Gelatin Zymography in Nanotoxicity Research.

明胶酶谱法在纳米毒性研究中的应用。

• DOI: 10.1007/978-1-4939-8916-4_8
• 发表时间: 2019
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Zhang,Yue;Wan,Rong;Zhang,Qunwei;Mo,Yiqun
• 通讯作者: Mo,Yiqun