Journal of Shandong University(Engineering Science) ›› 2019, Vol. 49 ›› Issue (3): 120-128.doi: 10.6040/j.issn.1672-3961.0.2018.443

• Chemistry and Environment • Previous Articles    

Toxic effects and mechanisms of oxidative stress induced by modified ultrafine carbon black

Chenhao JIA(),Rutao LIU*()   

  1. School of Environment Science and Engineering, Shandong University, Qingdao 266200, Shandong, China
  • Received:2018-10-18 Online:2019-06-20 Published:2019-06-27
  • Contact: Rutao LIU E-mail:1272466157@qq.com;rutaoliu@sdu.edu.cn
  • Supported by:
    国家自然科学基金资助项目(21477067);国家自然科学基金资助项目(21777088);高等学校博士学科点专项科研基金资助项目(708058)

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Abstract:

To study the toxic effects and mechanisms of oxidative stress induced by modified ultrafine carbon black (MCB), mouse hepatocytes and catalase (CAT) were exposed to MCB solutions. The cytotoxicity of MCB was assessed by CCK-8 kit, malondialdehyde (MDA) kit and CAT activity assay. The effects on CAT structure and function of MCB were investigated by utilizing fluorescence, UV—vis absorption and circular dichroism spectroscopy. The experiment results demonstrated that the hepatocyte viability decreased with the increase of MCB concentration. Low MCB doses (< 30 mg/L) increased CAT activity to protectcells from oxidative damage while high doses of MCB (> 30 mg/L) caused accumulation of MDA and redox imbalance in the cells, which induced oxidative damage in the liver. Spectroscopy studies found that MCB destroyed the secondary and tertiary structure of CAT and changed the microenvironment of amino acids, which made denaturation of the peptide chain. As a result, changes of skeleton structure reduced the activity of CAT. This study clarified the oxidative mechanism of MCB causing oxidative stress effects and provided a reference for the toxicity mechanism of nanomaterials.

Key words: modified ultrafine carbon black, oxidative stress, liver cells, catalase, fluorescence spectroscopy, ultraviolet-visible absorption spectroscopy

CLC Number: 

  • X503.1

Fig.1

TEM images and SEM images of MCB"

Fig.2

Dispersion of SB-100 and MCB in PB buffer"

Fig.3

Relative cell viability of hepatocytes under different exposure conditions"

Fig.4

Dose-effect fitting curve of cell viability as a function of MCB exposure concentration"

Fig.5

MDA contents in hepatocytes in the presence of MCB"

Fig.6

CAT relative activity exposed to different concentrations of MCB after 24 hours"

Fig.7

Effect of MCB on CAT fluorescence spectra"

Fig.8

UV-visible absorption spectra of CAT"

Fig.9

Schematic diagram for the binding of MCB with CAT"

Fig.10

Three-dimensional fluorescence spectrum of MCB and CAT"

Table 1

Three-dimensional characteristics of CAT combined with MCB"

MCB暴露的质量浓度/(mg·L-1) 峰1 峰2
激发波长/发射波长(nm/nm) 荧光强度 激发波长/发射波长(nm/nm) 荧光强度
0 280/280 4 309.0 280/326 5 600.3
2.5 280/284 9 035.4 280/328 5 006.1
5 280/284 9 078.6 280/328 3 862.7
10 280/284 9 595.5 280/330 2 633.3
15 280/284 9 615.8 280/330 1 493.1
20 280/284 9 680.5 280/332 1 040.7

Table 2

Effects of MCB on the secondary structure of CAT"

MCB暴露的质量浓度/ (mg·L-1) 二级结构含量/%
α-螺旋 β-折叠 β-转角 无序结构
0 43.4 18.8 14.4 23.1
5 45.8 18.0 14.2 22.9
10 47.9 17.4 13.9 21.6
20 49.5 16.7 13.5 21.3

Fig.11

CD spectra of CAT-MCB system"

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