多壁碳纳米管（MWCNTs）对体外人静脉内皮细胞（HUVECs）的不良血管效应：MWCNTs 长度的作用。
The adverse vascular effects of multi-walled carbon nanotubes (MWCNTs) to human vein endothelial cells (HUVECs) in vitro: role of length of MWCNTs.
机构信息
Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha, 410205, People's Republic of China.
Key Laboratory of Environment-Friendly Chemistry and Application of Ministry of Education, Lab of Biochemistry, College of Chemistry, Xiangtan University, Xiangtan, 411105, People's Republic of China.
出版信息
J Nanobiotechnology. 2017 Nov 10;15(1):80. doi: 10.1186/s12951-017-0318-x.
BACKGROUND
Increasing evidences indicate that exposure to multi-walled carbon nanotubes (MWCNTs) could induce adverse vascular effects, but the role of length of MWCNTs in determining the toxic effects is less studied. This study investigated the adverse effects of two well-characterized MWCNTs to human umbilical vein endothelial cells (HUVECs).
METHODS
The internalization and localization of MWCNTs in HUVECs were examined by using transmission electron microscopy (TEM). The cytotoxicity of MWCNTs to HUVECs was assessed by water soluble tetrazolium-8 (WST-8), lactate dehydrogenase (LDH) and neutral red uptake assays. Oxidative stress was indicated by the measurement of intracellular glutathione (GSH) and reactive oxygen species (ROS). ELISA was used to determine the release of inflammatory cytokines. THP-1 monocyte adhesion to HUVECs was also measured. To indicate the activation of endoplasmic reticulum (ER) stress, the expression of ddit3 and xbp-1s was measured by RT-PCR, and BiP protein level was measured by Western blot.
RESULTS
Transmission electron microscopy observation indicates the internalization of MWCNTs into HUVECs, with a localization in nuclei and mitochondria. The longer MWCNTs induced a higher level of cytotoxicity to HUVECs compared with the shorter ones. Neither of MWCNTs significantly promoted intracellular ROS, but the longer MWCNTs caused a higher depletion of GSH. Exposure to both types of MWCNTs significantly promoted THP-1 adhesion to HUVECs, accompanying with a significant increase of release of interleukin-6 (IL-6) but not tumor necrosis factor α (TNFα), soluble ICAM-1 (sICAM-1) or soluble VCAM-1 (sVCAM-1). Moreover, THP-1 adhesion and release of IL-6 and sVCAM-1 induced by the longer MWCNTs were significantly higher compared with the responses induced by the shorter ones. The biomarker of ER stress, ddit3 expression, but not xbp-1s expression or BiP protein level, was significantly induced by the exposure of longer MWCNTs.
CONCLUSIONS
Combined, these results indicated length dependent toxic effects of MWCNTs to HUVECs in vitro, which might be associated with oxidative stress and activation of ER stress.
背景
越来越多的证据表明，多壁碳纳米管（MWCNTs）的暴露会引起不良的血管效应，但MWCNTs 长度在确定其毒性效应中的作用研究较少。本研究调查了两种经过良好表征的 MWCNTs 对人脐静脉内皮细胞（HUVECs）的不良影响。
方法
使用透射电子显微镜（TEM）观察 MWCNTs 在 HUVECs 中的内化和定位。通过水溶性四唑盐-8（WST-8）、乳酸脱氢酶（LDH）和中性红摄取测定法评估 MWCNTs 对 HUVECs 的细胞毒性。通过测量细胞内谷胱甘肽（GSH）和活性氧（ROS）来指示氧化应激。ELISA 用于测定炎症细胞因子的释放。还测量了 THP-1 单核细胞与 HUVECs 的黏附。为了表明内质网（ER）应激的激活，通过 RT-PCR 测量 ddit3 和 xbp-1s 的表达，通过 Western blot 测量 BiP 蛋白水平。
结果
透射电子显微镜观察表明，MWCNTs 被内化到 HUVECs 中，并定位于细胞核和线粒体中。与较短的 MWCNTs 相比，较长的 MWCNTs 诱导了更高水平的 HUVECs 细胞毒性。两种 MWCNTs 都没有显著促进细胞内 ROS，但较长的 MWCNTs 导致 GSH 耗竭更高。两种类型的 MWCNTs 暴露均显著促进 THP-1 与 HUVECs 的黏附，伴随着白细胞介素-6（IL-6）的显著释放，但肿瘤坏死因子-α（TNFα）、可溶性细胞间黏附分子-1（sICAM-1）或可溶性血管细胞黏附分子-1（sVCAM-1）的释放没有显著增加。此外，较长的 MWCNTs 诱导的 THP-1 黏附和 IL-6 和 sVCAM-1 的释放明显高于较短的 MWCNTs 诱导的黏附和释放。内质网应激的生物标志物，ddit3 表达，但不是 xbp-1s 表达或 BiP 蛋白水平，由较长的 MWCNTs 暴露显著诱导。
结论
综上所述，这些结果表明 MWCNTs 对体外 HUVECs 的长度依赖性毒性作用，这可能与氧化应激和内质网应激的激活有关。
Zotero 插件