Microwave radiation affects the phosphorylation of nuclear factor erythroid-2-related factor-2 and activity of protein kinase C in vascular endothelial cells

Min Zhao, Yang Chen, Guang Bin Zhang, Zhen Ping Yu

Research output: Contribution to journalArticle

Abstract

Background: Nuclear factor erythroid-2-related factor-2 (NF-E2-related factor-2) is an important transcription factor to regulate anti-oxidative stress reaction. Some researches indicate that NF-E2-related factor-2 can be phosphorylated by numerous members of protein kinase C family. In order to investigate generant mechanism of microwave radiation on oxidative stress injury, whether microwave radiation can influence on anti-oxidative regulating system through NF-E2-related factor-2 or not should be further studied. Objective: To analyze the effect of microwave radiation on phosphorylation of NF-E2-related factor-2 and activity of protein kinase C in vascular endothelial cells. Design: Observational-contrast study. Setting: Department of Labor Hygiene, the Third Military Medical University of Chinese PLA. Materials: Vascular endothelial cell strain; H3 32PO4; Protein-A Sepharose (Sigma Company); mono-antibody of NF-E2-related factor-2 (H-300, Santa Cruz); α-mono-antibody of protein kinase C (Santa Cruz); glass microfiber filters (Whatman Company); gel scanning system (Gel Doc 2000, Bio-Rad); liquid scintillation spectrometer (LKB-117, Sweden). Methods: The experiment was carried out in Laboratory of Electromagnetic radiation and Biological Effect, Department of Labor Hygiene, the Third Military Medical University of Chinese PLA from March to July 2003. 1 Analysis of phosphorylation of NF-E2-related factor-2: Vascular endothelial cells were cultured with DMEM medium till the period of productive growth and incubated with 32Pi for 2 hours. And then, cultured bottle was maintained in water bath at 37°C and performed with microwave radiation in dark chamber, whose reflectivity was about zero. It was regarded as radiation group, and the average power density of radiation was 30 mW/cm2; in addition, the duration of radiation was 30 minutes. Cells did not deal with microwave radiation were regarded as control group. Phosphorylation level of NF-E2-related factor-2 was measured at 2, 4, 8 and 24 hours after radiation with immune coprecipitation-autoradiography technique and dealt with semi-quantitative analysis with gel scanning system. Cells in the control group were analyzed directly. 2 Active analysis and expressional measurement of protein kinase C: Cells in the radiation group and the control group were dealt with the same cultured method, condition, radiation styles, dosage and environment as mentioned above. At 2, 4, 8 and 24 hours after radiation, cells were split to extract plasma and membrane protein. Furthermore, activity of protein kinase C was measured with r-32P-ATP labeled liquid scintillation spectrometer; gray value of protein strap was dealt with semi-quantitative analysis with gel scanning system; staining degree of plasma was observed after immunocytochemical staining of protein kinase C. In addition, cells in the control group were measured and observed directly. Main outcome measures: 1 Phosphorylation level of NF-E2-related factor-2 in radiation group and control group; 2 Results of active analysis and expressional measurement of protein kinase C in radiation group and control group. Results: 1 Phosphorylation level of NF-E2-related factor-2 in radiation group and control group: Gray value of NF-E2-related factor-2 was higher in radiation group than that in control group at 2, 4 and 8 hours after radiation. Phosphorylation level of NF-E2-related factor-2 reached the peak at four hours after radiation. In addition, results of semi-quantitative scanning analysis showed that, at 2, 4 and 8 hours after radiation, phosphorytation level of NF-E2-related factor-2 was increased 33%, 261% and 141% in radiation group as compared with that in control group, respectively (t = 2.974, 4.209, 4.047, P < 0.05), and then, fallen down to normal value 24 hours later. 2 Results of active analysis and expressional measurement of protein kinase C in radiation group and control group: At 2, 4 and 8 hours after microwave radiation, expression of protein kinase C in radiation group was higher than that in control group, especially at the 4 hour. In addition, at 24 hours after radiation, expression of protein kinase C recovered the normal value. Results of immunocytochemical staining showed that staining of plasma was deeper in radiation group than that in control group at 4 hours after radiation. Moreover, results of r-32P-ATP labeled liquid scintillation spectrometer also suggested that, at 2, 4 and 8 hours after radiation, activity of protein kinase C was increased 36%, 93% and 47% in radiation group as compared with that in control group, respectively (t=2.801, 3.654, 3.035, P < 0.05). And then, activity of protein kinase C was decreased after 24 hours, otherwise, activity of protein kinase C reached the peak at 4 hours after radiation. Conclusion: Microwave radiation can strengthen the phosphorylation level of NF-E2-related factor-2 in vascular endothelial cells during a special period; meanwhile, it can also cause the increase of expression of protein kinase C. Time effect of activity of protein kinase C is coincidence with phosphorylation level of NF-E2-related factor-2.

Original languageEnglish (US)
Pages (from-to)2765-2768
Number of pages4
JournalJournal of Clinical Rehabilitative Tissue Engineering Research
Volume11
Issue number14
StatePublished - Apr 8 2007
Externally publishedYes

Fingerprint

Phosphorylation
Endothelial cells
Microwaves
Protein Kinase C
NF-E2-Related Factor 2
Endothelial Cells
Radiation
Proteins
Control Groups
Scintillation counters
Gels
Military Hygiene
Scanning
Oxidative stress
Staining and Labeling
Adenosinetriphosphate
Plasmas
Antibodies
Radiation Effects
Liquids

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biomedical Engineering
  • Transplantation

Cite this

@article{84d4686c259c48bfb100d02aab06d950,
title = "Microwave radiation affects the phosphorylation of nuclear factor erythroid-2-related factor-2 and activity of protein kinase C in vascular endothelial cells",
abstract = "Background: Nuclear factor erythroid-2-related factor-2 (NF-E2-related factor-2) is an important transcription factor to regulate anti-oxidative stress reaction. Some researches indicate that NF-E2-related factor-2 can be phosphorylated by numerous members of protein kinase C family. In order to investigate generant mechanism of microwave radiation on oxidative stress injury, whether microwave radiation can influence on anti-oxidative regulating system through NF-E2-related factor-2 or not should be further studied. Objective: To analyze the effect of microwave radiation on phosphorylation of NF-E2-related factor-2 and activity of protein kinase C in vascular endothelial cells. Design: Observational-contrast study. Setting: Department of Labor Hygiene, the Third Military Medical University of Chinese PLA. Materials: Vascular endothelial cell strain; H3 32PO4; Protein-A Sepharose (Sigma Company); mono-antibody of NF-E2-related factor-2 (H-300, Santa Cruz); α-mono-antibody of protein kinase C (Santa Cruz); glass microfiber filters (Whatman Company); gel scanning system (Gel Doc 2000, Bio-Rad); liquid scintillation spectrometer (LKB-117, Sweden). Methods: The experiment was carried out in Laboratory of Electromagnetic radiation and Biological Effect, Department of Labor Hygiene, the Third Military Medical University of Chinese PLA from March to July 2003. 1 Analysis of phosphorylation of NF-E2-related factor-2: Vascular endothelial cells were cultured with DMEM medium till the period of productive growth and incubated with 32Pi for 2 hours. And then, cultured bottle was maintained in water bath at 37°C and performed with microwave radiation in dark chamber, whose reflectivity was about zero. It was regarded as radiation group, and the average power density of radiation was 30 mW/cm2; in addition, the duration of radiation was 30 minutes. Cells did not deal with microwave radiation were regarded as control group. Phosphorylation level of NF-E2-related factor-2 was measured at 2, 4, 8 and 24 hours after radiation with immune coprecipitation-autoradiography technique and dealt with semi-quantitative analysis with gel scanning system. Cells in the control group were analyzed directly. 2 Active analysis and expressional measurement of protein kinase C: Cells in the radiation group and the control group were dealt with the same cultured method, condition, radiation styles, dosage and environment as mentioned above. At 2, 4, 8 and 24 hours after radiation, cells were split to extract plasma and membrane protein. Furthermore, activity of protein kinase C was measured with r-32P-ATP labeled liquid scintillation spectrometer; gray value of protein strap was dealt with semi-quantitative analysis with gel scanning system; staining degree of plasma was observed after immunocytochemical staining of protein kinase C. In addition, cells in the control group were measured and observed directly. Main outcome measures: 1 Phosphorylation level of NF-E2-related factor-2 in radiation group and control group; 2 Results of active analysis and expressional measurement of protein kinase C in radiation group and control group. Results: 1 Phosphorylation level of NF-E2-related factor-2 in radiation group and control group: Gray value of NF-E2-related factor-2 was higher in radiation group than that in control group at 2, 4 and 8 hours after radiation. Phosphorylation level of NF-E2-related factor-2 reached the peak at four hours after radiation. In addition, results of semi-quantitative scanning analysis showed that, at 2, 4 and 8 hours after radiation, phosphorytation level of NF-E2-related factor-2 was increased 33{\%}, 261{\%} and 141{\%} in radiation group as compared with that in control group, respectively (t = 2.974, 4.209, 4.047, P < 0.05), and then, fallen down to normal value 24 hours later. 2 Results of active analysis and expressional measurement of protein kinase C in radiation group and control group: At 2, 4 and 8 hours after microwave radiation, expression of protein kinase C in radiation group was higher than that in control group, especially at the 4 hour. In addition, at 24 hours after radiation, expression of protein kinase C recovered the normal value. Results of immunocytochemical staining showed that staining of plasma was deeper in radiation group than that in control group at 4 hours after radiation. Moreover, results of r-32P-ATP labeled liquid scintillation spectrometer also suggested that, at 2, 4 and 8 hours after radiation, activity of protein kinase C was increased 36{\%}, 93{\%} and 47{\%} in radiation group as compared with that in control group, respectively (t=2.801, 3.654, 3.035, P < 0.05). And then, activity of protein kinase C was decreased after 24 hours, otherwise, activity of protein kinase C reached the peak at 4 hours after radiation. Conclusion: Microwave radiation can strengthen the phosphorylation level of NF-E2-related factor-2 in vascular endothelial cells during a special period; meanwhile, it can also cause the increase of expression of protein kinase C. Time effect of activity of protein kinase C is coincidence with phosphorylation level of NF-E2-related factor-2.",
author = "Min Zhao and Yang Chen and Zhang, {Guang Bin} and Yu, {Zhen Ping}",
year = "2007",
month = "4",
day = "8",
language = "English (US)",
volume = "11",
pages = "2765--2768",
journal = "Chinese Journal of Tissue Engineering Research",
issn = "1673-8225",
publisher = "Journal of Clinical Rehabilitative Tissue Engineering Research",
number = "14",

}

TY - JOUR

T1 - Microwave radiation affects the phosphorylation of nuclear factor erythroid-2-related factor-2 and activity of protein kinase C in vascular endothelial cells

AU - Zhao, Min

AU - Chen, Yang

AU - Zhang, Guang Bin

AU - Yu, Zhen Ping

PY - 2007/4/8

Y1 - 2007/4/8

N2 - Background: Nuclear factor erythroid-2-related factor-2 (NF-E2-related factor-2) is an important transcription factor to regulate anti-oxidative stress reaction. Some researches indicate that NF-E2-related factor-2 can be phosphorylated by numerous members of protein kinase C family. In order to investigate generant mechanism of microwave radiation on oxidative stress injury, whether microwave radiation can influence on anti-oxidative regulating system through NF-E2-related factor-2 or not should be further studied. Objective: To analyze the effect of microwave radiation on phosphorylation of NF-E2-related factor-2 and activity of protein kinase C in vascular endothelial cells. Design: Observational-contrast study. Setting: Department of Labor Hygiene, the Third Military Medical University of Chinese PLA. Materials: Vascular endothelial cell strain; H3 32PO4; Protein-A Sepharose (Sigma Company); mono-antibody of NF-E2-related factor-2 (H-300, Santa Cruz); α-mono-antibody of protein kinase C (Santa Cruz); glass microfiber filters (Whatman Company); gel scanning system (Gel Doc 2000, Bio-Rad); liquid scintillation spectrometer (LKB-117, Sweden). Methods: The experiment was carried out in Laboratory of Electromagnetic radiation and Biological Effect, Department of Labor Hygiene, the Third Military Medical University of Chinese PLA from March to July 2003. 1 Analysis of phosphorylation of NF-E2-related factor-2: Vascular endothelial cells were cultured with DMEM medium till the period of productive growth and incubated with 32Pi for 2 hours. And then, cultured bottle was maintained in water bath at 37°C and performed with microwave radiation in dark chamber, whose reflectivity was about zero. It was regarded as radiation group, and the average power density of radiation was 30 mW/cm2; in addition, the duration of radiation was 30 minutes. Cells did not deal with microwave radiation were regarded as control group. Phosphorylation level of NF-E2-related factor-2 was measured at 2, 4, 8 and 24 hours after radiation with immune coprecipitation-autoradiography technique and dealt with semi-quantitative analysis with gel scanning system. Cells in the control group were analyzed directly. 2 Active analysis and expressional measurement of protein kinase C: Cells in the radiation group and the control group were dealt with the same cultured method, condition, radiation styles, dosage and environment as mentioned above. At 2, 4, 8 and 24 hours after radiation, cells were split to extract plasma and membrane protein. Furthermore, activity of protein kinase C was measured with r-32P-ATP labeled liquid scintillation spectrometer; gray value of protein strap was dealt with semi-quantitative analysis with gel scanning system; staining degree of plasma was observed after immunocytochemical staining of protein kinase C. In addition, cells in the control group were measured and observed directly. Main outcome measures: 1 Phosphorylation level of NF-E2-related factor-2 in radiation group and control group; 2 Results of active analysis and expressional measurement of protein kinase C in radiation group and control group. Results: 1 Phosphorylation level of NF-E2-related factor-2 in radiation group and control group: Gray value of NF-E2-related factor-2 was higher in radiation group than that in control group at 2, 4 and 8 hours after radiation. Phosphorylation level of NF-E2-related factor-2 reached the peak at four hours after radiation. In addition, results of semi-quantitative scanning analysis showed that, at 2, 4 and 8 hours after radiation, phosphorytation level of NF-E2-related factor-2 was increased 33%, 261% and 141% in radiation group as compared with that in control group, respectively (t = 2.974, 4.209, 4.047, P < 0.05), and then, fallen down to normal value 24 hours later. 2 Results of active analysis and expressional measurement of protein kinase C in radiation group and control group: At 2, 4 and 8 hours after microwave radiation, expression of protein kinase C in radiation group was higher than that in control group, especially at the 4 hour. In addition, at 24 hours after radiation, expression of protein kinase C recovered the normal value. Results of immunocytochemical staining showed that staining of plasma was deeper in radiation group than that in control group at 4 hours after radiation. Moreover, results of r-32P-ATP labeled liquid scintillation spectrometer also suggested that, at 2, 4 and 8 hours after radiation, activity of protein kinase C was increased 36%, 93% and 47% in radiation group as compared with that in control group, respectively (t=2.801, 3.654, 3.035, P < 0.05). And then, activity of protein kinase C was decreased after 24 hours, otherwise, activity of protein kinase C reached the peak at 4 hours after radiation. Conclusion: Microwave radiation can strengthen the phosphorylation level of NF-E2-related factor-2 in vascular endothelial cells during a special period; meanwhile, it can also cause the increase of expression of protein kinase C. Time effect of activity of protein kinase C is coincidence with phosphorylation level of NF-E2-related factor-2.

AB - Background: Nuclear factor erythroid-2-related factor-2 (NF-E2-related factor-2) is an important transcription factor to regulate anti-oxidative stress reaction. Some researches indicate that NF-E2-related factor-2 can be phosphorylated by numerous members of protein kinase C family. In order to investigate generant mechanism of microwave radiation on oxidative stress injury, whether microwave radiation can influence on anti-oxidative regulating system through NF-E2-related factor-2 or not should be further studied. Objective: To analyze the effect of microwave radiation on phosphorylation of NF-E2-related factor-2 and activity of protein kinase C in vascular endothelial cells. Design: Observational-contrast study. Setting: Department of Labor Hygiene, the Third Military Medical University of Chinese PLA. Materials: Vascular endothelial cell strain; H3 32PO4; Protein-A Sepharose (Sigma Company); mono-antibody of NF-E2-related factor-2 (H-300, Santa Cruz); α-mono-antibody of protein kinase C (Santa Cruz); glass microfiber filters (Whatman Company); gel scanning system (Gel Doc 2000, Bio-Rad); liquid scintillation spectrometer (LKB-117, Sweden). Methods: The experiment was carried out in Laboratory of Electromagnetic radiation and Biological Effect, Department of Labor Hygiene, the Third Military Medical University of Chinese PLA from March to July 2003. 1 Analysis of phosphorylation of NF-E2-related factor-2: Vascular endothelial cells were cultured with DMEM medium till the period of productive growth and incubated with 32Pi for 2 hours. And then, cultured bottle was maintained in water bath at 37°C and performed with microwave radiation in dark chamber, whose reflectivity was about zero. It was regarded as radiation group, and the average power density of radiation was 30 mW/cm2; in addition, the duration of radiation was 30 minutes. Cells did not deal with microwave radiation were regarded as control group. Phosphorylation level of NF-E2-related factor-2 was measured at 2, 4, 8 and 24 hours after radiation with immune coprecipitation-autoradiography technique and dealt with semi-quantitative analysis with gel scanning system. Cells in the control group were analyzed directly. 2 Active analysis and expressional measurement of protein kinase C: Cells in the radiation group and the control group were dealt with the same cultured method, condition, radiation styles, dosage and environment as mentioned above. At 2, 4, 8 and 24 hours after radiation, cells were split to extract plasma and membrane protein. Furthermore, activity of protein kinase C was measured with r-32P-ATP labeled liquid scintillation spectrometer; gray value of protein strap was dealt with semi-quantitative analysis with gel scanning system; staining degree of plasma was observed after immunocytochemical staining of protein kinase C. In addition, cells in the control group were measured and observed directly. Main outcome measures: 1 Phosphorylation level of NF-E2-related factor-2 in radiation group and control group; 2 Results of active analysis and expressional measurement of protein kinase C in radiation group and control group. Results: 1 Phosphorylation level of NF-E2-related factor-2 in radiation group and control group: Gray value of NF-E2-related factor-2 was higher in radiation group than that in control group at 2, 4 and 8 hours after radiation. Phosphorylation level of NF-E2-related factor-2 reached the peak at four hours after radiation. In addition, results of semi-quantitative scanning analysis showed that, at 2, 4 and 8 hours after radiation, phosphorytation level of NF-E2-related factor-2 was increased 33%, 261% and 141% in radiation group as compared with that in control group, respectively (t = 2.974, 4.209, 4.047, P < 0.05), and then, fallen down to normal value 24 hours later. 2 Results of active analysis and expressional measurement of protein kinase C in radiation group and control group: At 2, 4 and 8 hours after microwave radiation, expression of protein kinase C in radiation group was higher than that in control group, especially at the 4 hour. In addition, at 24 hours after radiation, expression of protein kinase C recovered the normal value. Results of immunocytochemical staining showed that staining of plasma was deeper in radiation group than that in control group at 4 hours after radiation. Moreover, results of r-32P-ATP labeled liquid scintillation spectrometer also suggested that, at 2, 4 and 8 hours after radiation, activity of protein kinase C was increased 36%, 93% and 47% in radiation group as compared with that in control group, respectively (t=2.801, 3.654, 3.035, P < 0.05). And then, activity of protein kinase C was decreased after 24 hours, otherwise, activity of protein kinase C reached the peak at 4 hours after radiation. Conclusion: Microwave radiation can strengthen the phosphorylation level of NF-E2-related factor-2 in vascular endothelial cells during a special period; meanwhile, it can also cause the increase of expression of protein kinase C. Time effect of activity of protein kinase C is coincidence with phosphorylation level of NF-E2-related factor-2.

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M3 - Article

VL - 11

SP - 2765

EP - 2768

JO - Chinese Journal of Tissue Engineering Research

JF - Chinese Journal of Tissue Engineering Research

SN - 1673-8225

IS - 14

ER -