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

基本信息

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
项目类别：
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

项目摘要

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，正在开发大量金属纳米颗粒，并以新的配方和表面特性生产以满足新需求。例如，很重要过渡金属，镍纳米颗粒（纳米-NI）在电池领域具有广泛的应用，电导体，永久磁铁，磁性流体，磁记录介质，太阳能吸收，燃料电池电极和催化剂。另外，由于结构和固有的化学和物理物业，镍合金纳米材料已在生物医学应用中特别感兴趣。作为使用纳米材料不断增长，纳米颗粒污染环境污染的风险正在增加。这术语“纳米毒理学”已被用来提出以下概念，即纳米颗粒可能代表了独特与常规致病颗粒不同的颗粒毒素类别。在此提案中，我们选择了纳米-NI作为型号金属纳米颗粒，因为其工业兴趣广泛，生物学和医学申请。根据我们的初步数据，我们假设纳米-NI将激活单核细胞/巨噬细胞改变基质金属蛋白酶（MMP）的表达和活性通过低氧诱导因子1a（HIF-1A）和microRNA miR-21。在结构上可能量身定制的含Ni的纳米颗粒（例如镍合金纳米颗粒）可能更少甚至没有效果。知道这将非常重要。该项目将同时使用体外和体内系统来解决以下目标。（1）。确定MMP和TIMPS表达和活动的改变在体外和体内暴露于纳米-NI或镍合金纳米颗粒的单核细胞/巨噬细胞中。我们将检查对纳米-NI的接触是否会改变MMP-2，MMP-9，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）和从小鼠分离的单核细胞，通过使用体外定量实时PCR，明胶Zymography或反向Zymography测定法。我们还将确定从纳米-NI-i-pos小鼠分离的单核细胞中MMP和TIMP的表达和活性的改变。我们将研究镍合金纳米颗粒，纳米ni核心和二氧化硅或碳壳，以及二氧化硅或二氧化硅或碳支持的纳米-NI将减少纳米ni诱导的MMP的激活，并在体外和IN中降低毒性效应体内设置。（2）。检查暴露于纳米-NI的单核细胞/巨噬细胞中的HIF-1A积累 HIF-1A对纳米-NI诱导的MMPs表达和活性改变的作用。我们将暴露于纳米-NI的情况下，测量U937细胞中的HIF-1A水平。研究HIF-1A在纳米ni中的作用诱导MMP的表达和活动，我们将采用以下策略：（1）使用HIF-1A抑制剂例如热冲蛋白90（HSP90）；（2）使用HIF-1A特异性siRNA敲除hif-1a表达；（3）使用HIF-1A敲除细胞。（3）。调查纳米ni诱导的MMP的改变通过激活HIF-1A的microRNA miR-21上调，表达和活性。我们将首先确定暴露于纳米-NI是否会通过定量实时PCR引起miR-21的上调。我们将然后研究miR-21是否通过使用：（1）通过激活HIF-1A调节MMP的表达和活性 MIR-21前转染以增加细胞中miR-21的丰度；（2）抗MIR-21转染至较低细胞中miR-21的水平；（3）使用HIF诱饵阻止内源性HIF转录的活性细胞中的因素。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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.)
影响因子：
0
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.)
影响因子：
0
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。