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Human selenium-containing single-chain variable fragment with glutathione peroxidase activity protects NIH3T3 fibroblast against oxidative damage

  • Molecular Cell Biology
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Abstract

Ultraviolet B (UVB medium wave, 280–315 nm) induces cellular oxidative damage and apoptosis by producing reactive oxygen species (ROS). Glutathione peroxidase functions as an antioxidant by catalyzing the reduction of hydrogen peroxide, the more important member of reactive oxygen species. A human selenium-containing single-chain variable fragment (se-scFv-B3) with glutathione peroxidase activity of 1288 U/μmol was generated and investigated for its antioxidant effects in UVB-induced oxidative damage model. In particular, cell viability, lipid peroxidation extent, cell apoptosis, the change of mitochondrial membrane potential, caspase-3 activity and the levels of intracellular reactive oxygen species were assayed. Human se-scFv-B3 protects NIH3T3 cells against ultraviolet B-induced oxidative damage and subsequent apoptosis by prevention of lipid peroxidation, inhibition of the collapse of mitochondrial membrane potential as well as the suppression of the caspase-3 activity and the level of intracellular ROS. It seems that antioxidant effects of human se-scFv-B3 are mainly associated with its capability to scavenge reactive oxygen species, which is similar to that of the natural glutathione peroxidase.

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Abbreviations

ROS:

reactive oxygen species

H2O2 :

hydrogen peroxide

GPX:

Glutathione peroxidase

UVB:

ultraviolet B

se-scFv:

selenium-containing single-chain variable fragment

MDA:

malondialdehyde

DiOC6(3):

3,3-dihexyloxacarbocyanine iodide

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

ΔΨm :

mitochondrial membrane potential

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Authors and Affiliations

  1. College of Pharmaceutical Science, Jilin University, Changchun, 130021, P. R. China

    R. Huo & J. Y. Wei

  2. China-Japan Union Hospital, Jilin University, Changchun, 130033, P. R. China

    L. Yang

  3. College of Physical Education, Jilin University, Changchun, 130021, P. R. China

    T. G. Zhang

Authors
  1. R. Huo
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  2. L. Yang
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  3. T. G. Zhang
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  4. J. Y. Wei
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Corresponding author

Correspondence to J. Y. Wei.

Additional information

Published in Russian in Molekulyarnaya Biologiya, 2017, Vol. 51, No. 3, pp. 483–489.

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Huo, R., Yang, L., Zhang, T.G. et al. Human selenium-containing single-chain variable fragment with glutathione peroxidase activity protects NIH3T3 fibroblast against oxidative damage. Mol Biol 51, 426–431 (2017). https://doi.org/10.1134/S0026893317030074

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  • DOI: https://doi.org/10.1134/S0026893317030074

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