Evaluation of UVA-induced oxidative stress using a highly sensitive chemiluminescence method

Oxidative stress is mainly mediated by reactive oxygen species (ROS). Evaluation of oxidative stress is helpful for choosing an appropriate method to protect the organism from the oxidative damage. In this study, a highly sensitive and simple chemiluminescence method is presented for the evaluation of radiation-induced oxidative stress in human peripheral lymphocytes. The lymphocytes were irradiated by ultraviolet radiation (320-400nm, UVA) with different doses. The ROS generated by the lymphocytes was detected by chemiluminescence method, using a highly sensitive chemiluminescence probe 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-α] pyrazin-3-one (MCLA). The cell viability was detected with Cell Counting Kit-8 (CCK-8). The malondialdehyde (MDA), a marker of lipid peroxidation and oxidative stress, and the total antioxidant capacity (TAC), a parameter that is taken as evidence of oxidative stress, were measured too. The results show that both chemiluminescence intensity, cell mortality and MDA concentration of lymphocytes grow with the increase of UVA dose range from 0.5 to 8 J/cm2, while the TAC decreases. There exists a positive relationship between cell oxidative damage degree and the chemiluminescence intensity of lymphocytes. This highly sensitive chemiluminescence method would potentially provide an easy way to evaluate the level of UVA-induced oxidative stress readily, sensitively and rapidly

[1]  R. Ardaillou,et al.  Reactive oxygen species: production and role in the kidney. , 1986, The American journal of physiology.

[2]  Nikolaos Gkantidis,et al.  Clinical Significance , 2022, The SAGE Encyclopedia of Research Design.

[3]  上原 清,et al.  The first application of a chemiluminescence probe, 2-methyl-6-[p-methoxyphenyl]-3,7-dihydroimidazo[1,2-a]pyrazin-3-one (MCLA), for detecting O[2[-] production, in vitro : from Kupffer cells stimulated by phorbol myristate acetate , 1994 .

[4]  F. J. Romero,et al.  Lipid peroxidation products and antioxidants in human disease. , 1998, Environmental health perspectives.

[5]  Ze'ev Ronai,et al.  Role of redox potential and reactive oxygen species in stress signaling , 1999, Oncogene.

[6]  N. Dhalla,et al.  Role of oxidative stress in cardiovascular diseases , 2000, Journal of hypertension.

[7]  H. Sakurai,et al.  Chemiluminescent detection and imaging of reactive oxygen species in live mouse skin exposed to UVA. , 2000, Biochemical and biophysical research communications.

[8]  C. Catena,et al.  Micronucleus yield and colorimetric test as indicators of damage in patients' lymphocytes after 131I therapy. , 2000, Journal of nuclear medicine : official publication, Society of Nuclear Medicine.

[9]  Michael E. Phelps,et al.  Ex vivo cell labeling with 64Cu–pyruvaldehyde-bis(N4-methylthiosemicarbazone) for imaging cell trafficking in mice with positron-emission tomography , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[10]  Abraham Nyska,et al.  Invited Review: Oxidation of Biological Systems: Oxidative Stress Phenomena, Antioxidants, Redox Reactions, and Methods for Their Quantification , 2002, Toxicologic pathology.

[11]  X. Luan,et al.  CP27 affects viability, proliferation, attachment and gene expression in embryonic fibroblasts , 2002, Cell proliferation.

[12]  V. Sypin,et al.  [Estimation of genetic effects of chronic exposure to low-dose rate gamma-radiation by cytogenetic methods and DNA-comet assay]. , 2003, Radiatsionnaia biologiia, radioecologiia.

[13]  B. Shankar,et al.  Generation of Reactive Oxygen Species and Radiation Response in Lymphocytes and Tumor Cells , 2003, Radiation research.

[14]  A. Świerniak,et al.  Radiation-Induced Micronucleus Frequency in Peripheral Blood Lymphocytes is Correlated with Normal Tissue Damage in Patients with Cervical Carcinoma Undergoing Radiotherapy , 2003, Radiation research.

[15]  D. Bernhard,et al.  Enhanced MTT-reducing activity under growth inhibition by resveratrol in CEM-C7H2 lymphocytic leukemia cells. , 2003, Cancer letters.

[16]  Y. Ogawa,et al.  Radiation-induced reactive oxygen species formation prior to oxidative DNA damage in human peripheral T cells. , 2003, International journal of molecular medicine.

[17]  D. Xing,et al.  Imaging of ultra-weak bio-chemiluminescence and singlet oxygen generation in germinating soybean in response to wounding. , 2003, Luminescence : the journal of biological and chemical luminescence.

[18]  R. Touyz Reactive oxygen species, vascular oxidative stress, and redox signaling in hypertension: what is the clinical significance? , 2004, Hypertension.

[19]  M. Nakano,et al.  Detection of Active Oxygen Species in Biological Systems , 1998, Cellular and Molecular Neurobiology.

[20]  T. Neumann,et al.  Lysophosphatidylcholine-induced modulation of Ca(2+)-activated K(+)channels contributes to ROS-dependent proliferation of cultured human endothelial cells. , 2004, Journal of molecular and cellular cardiology.

[21]  J. Andersen,et al.  Oxidative stress in neurodegeneration: cause or consequence? , 2004, Nature Reviews Neuroscience.

[22]  B. Halliwell,et al.  Measuring reactive species and oxidative damage in vivo and in cell culture: how should you do it and what do the results mean? , 2004, British journal of pharmacology.

[23]  D. Xing,et al.  Evaluation of the degree of medical radiation damage with a highly sensitive chemiluminescence method. , 2004, Luminescence : the journal of biological and chemical luminescence.

[24]  G. Schmidt,et al.  Correction for Adonai et al., Ex vivo cell labeling with 64Cu-pyruvaldehyde-bis(N4-methylthiosemicarbazone) for imaging cell trafficking in mice with positron-emission tomography , 2006, Proceedings of the National Academy of Sciences.