Multi-walled carbon nanotubes induce COX-2 and iNOS expression via MAP kinase-dependent and -independent mechanisms in mouse RAW264.7 macrophages.
Multi-walled carbon nanotubes induce COX-2 and iNOS expression via MAP kinase-dependent and -independent mechanisms in mouse RAW264.7 macrophages.
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DOI：
10.1186/1743-8977-9-14
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发表时间：
2012-05-09
影响因子：
10
通讯作者：
Langenbach R
中科院分区：
文献类型：
--
作者：
Lee JK;Sayers BC;Chun KS;Lao HC;Shipley-Phillips JK;Bonner JC;Langenbach R
碳纳米管（CNT）是工程石墨烯圆柱体，在工程、电子和医学中具有许多应用。然而，CNT在啮齿动物肺部引起炎症和纤维化，表明潜在的人类健康风险。我们假设多壁碳纳米管（MWCNTs）通过激活细胞外信号调节激酶（ERK 1，2）诱导巨噬细胞中的两种关键炎症酶，环氧合酶-2（考克斯-2）和诱导型一氧化氮合酶（iNOS）。RAW264.7巨噬细胞在一定剂量和时间范围内暴露于MWCNT或炭黑纳米颗粒（CBNP）。多壁碳纳米管的摄取和亚细胞定位通过透射电子显微镜（TEM）可视化。通过Western印迹分析测量考克斯-2、iNOS和ERK 1，2（总ERK和磷酸化ERK）的蛋白水平。ELISA法检测细胞培养上清中前列腺素E2（PGE 2）的含量，Greiss法检测细胞培养上清中一氧化氮（NO）的含量。多壁碳纳米管，而不是CBNPs，诱导考克斯-2和iNOS的时间和剂量依赖性的方式。多壁碳纳米管诱导的考克斯-2和iNOS分别与PGE 2和NO产生的增加相关。多壁碳纳米管可激活ERK 1，2，抑制ERK 1，2（U 0126）可阻断多壁碳纳米管诱导的考克斯-2和PGE 2的产生，但不减少诱导型一氧化氮合酶的产生。抑制iNOS（L-NAME）不影响ERK 1，2激活，L-NAME也不显著降低MWCNT对考克斯-2的诱导。镍纳米颗粒（NiNPs）作为残留催化剂存在于MWCNT中，也通过ERK-1诱导考克斯-2，2。然而，通过比较含有4.5%和1.8% Ni的MWCNT对考克斯-2的诱导，未显示出诱导考克斯-2的能力的显著差异，表明除了Ni含量之外，MWCNT的特性也有助于考克斯-2的诱导。本研究确定考克斯-2和随后的PGE 2的生产，沿着诱导型一氧化氮合酶和NO的生产，作为炎症介质参与巨噬细胞对多壁碳纳米管的反应。此外，我们的工作表明，考克斯-2诱导的多壁碳纳米管在RAW 264.7巨噬细胞是ERK 1，2依赖性，而诱导型一氧化氮合酶诱导的多壁碳纳米管是ERK 1，2非依赖性。我们的数据还表明，除了残留的镍催化剂外，其他物理化学因素也在考克斯-2诱导成多壁碳纳米管中发挥作用。
Carbon nanotubes (CNTs) are engineered graphene cylinders with numerous applications in engineering, electronics and medicine. However, CNTs cause inflammation and fibrosis in the rodent lung, suggesting a potential human health risk. We hypothesized that multi-walled CNTs (MWCNTs) induce two key inflammatory enzymes in macrophages, cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS), through activation of extracellular signal-regulated kinases (ERK1,2). RAW264.7 macrophages were exposed to MWCNTs or carbon black nanoparticles (CBNPs) over a range of doses and time course. Uptake and subcellular localization of MWCNTs was visualized by transmission electron microscopy (TEM). Protein levels of COX-2, iNOS, and ERK1,2 (total ERK and phosphorylated ERK) were measured by Western blot analysis. Prostaglandin-E2 (PGE2) and nitric oxide (NO) levels in cell supernatants were measured by ELISA and Greiss assay, respectively. MWCNTs, but not CBNPs, induced COX-2 and iNOS in a time- and dose-dependent manner. COX-2 and iNOS induction by MWCNTs correlated with increased PGE2 and NO production, respectively. MWCNTs caused ERK1,2 activation and inhibition of ERK1,2 (U0126) blocked MWCNT induction of COX-2 and PGE2 production, but did not reduce the induction of iNOS. Inhibition of iNOS (L-NAME) did not affect ERK1,2 activation, nor did L-NAME significantly decrease COX-2 induction by MWCNT. Nickel nanoparticles (NiNPs), which are present in MWCNTs as a residual catalyst, also induced COX-2 via ERK-1,2. However, a comparison of COX-2 induction by MWCNTs containing 4.5 and 1.8% Ni did not show a significant difference in ability to induce COX-2, indicating that characteristics of MWCNTs in addition to Ni content contribute to COX-2 induction. This study identifies COX-2 and subsequent PGE2 production, along with iNOS induction and NO production, as inflammatory mediators involved in the macrophage response to MWCNTs. Furthermore, our work demonstrates that COX-2 induction by MWCNTs in RAW264.7 macrophages is ERK1,2-dependent, while iNOS induction by MWCNTs is ERK1,2-independent. Our data also suggest contributory physicochemical factors other than residual Ni catalyst play a role in COX-2 induction to MWCNT.
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影响因子：
38.3
作者：
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影响因子：
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DOI：
10.1165/rcmb.2008-0276oc
发表时间：
2009-03-01
影响因子：
6.4
作者：
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影响因子：
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