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Neurochemical Research

, Volume 38, Issue 7, pp 1315–1323 | Cite as

Genistein alleviates the mitochondria-targeted DNA damage induced by β-amyloid peptides 25–35 in C6 glioma cells

  • Wei-wei Ma
  • Cheng-cheng Hou
  • Xin Zhou
  • Huan-ling Yu
  • Yuan-di Xi
  • Juan Ding
  • Xia Zhao
  • Rong XiaoEmail author
Original Paper

Abstract

Reactive oxygen species (ROS) are mainly produced by mitochondria which can cause oxidative stress. It has been considered that mitochondrial damage induced by oxidative stress is related to Alzheimer’s disease (AD). Besides, mitochondrial DNA (mtDNA) is more vulnerable to oxidative damage than other biomacromolecules, causing serious dysfunction to mitochondria. β-amyloid peptides (Aβ) is a main factor responsible for the occurence and development of AD. Astrocytes is an important target cell for Aβ’ toxicity and can be activated to neglect their normal fountain in the central nervous system. Genistein (Gen), a main active ingredient of soybean isoflavone, has been shown to have neuroprotective effects by antagonizing oxidative damage induced by Aβ. Thus, in the present study, we evaluated Aβ25–35 induced mitochondrial DNA (mtDNA) damage and the protective effect of Gen in C6 glioma cells (C6 cells). The study design was consisted of four groups: control group (vehicle), Aβ group treated with Aβ25–35, Gen + Aβ group treated with Gen + Aβ25–35 and Gen group treated with Gen only. C6 cells were pre-incubated with or without Gen (50 μM) for 2 h followed by the incubation with Aβ25–35 (25 μM) for another 24 h. Then the cells were harvested and processed to perform the analysis according to protocols. The mitochondrial ROS in C6 cells were measured by fluorescence spectrometer. Enzyme-linked immunosorbent assay (ELISA) was used to detect the mitochondrial reduced glutathione (GSH) and oxidized glutathione (GSSG) in C6 cells, then the ratio of GSH and GSSG was calculated. The levels of 8-hydroxydeoxyguanosine (8-OHdG) in C6 cells was also detected by ELISA. In addition, mtDNA deletion was detected by polymerase chain reaction (PCR). The mRNA and protein expression of 8-oxoguanine DNA glycosylase (OGG1) in both C6 cells and its mitochondria, and manganese superoxide dismutase (MnSOD) in mitochondria were detected by using reverse transcription-PCR and Western blot. The results showed that the increased mitochondrial ROS accumulation in C6 cells induced by Aβ was profoundly reversed by pre-treaded with Gen (p < 0.05). The ratio of GSH and GSSG in mitochondria was significantly increased in both Gen + Aβ group and Gen group compared with Aβ group (p < 0.05). The levels of 8-OHdG in C6 cells and mtDNA deletion were decreased after pre-treated with Gen (p < 0.05). Gen could also up-regulate the mRNA and protein expression of OGG1 in both C6 cells and its mitochondria and mitochondrial MnSOD compared with the Aβ group (p < 0.05). These results confirmed that Gen could alleviate the mitochondria-targeted oxidative damage induced by β-amyloid 25–35 in C6 cells which might be useful for the treatment of neurodegenerative diseases.

Keywords

Genistein β-Amyloid peptides 25–35 Oxidative stress DNA damage Mitochondria C6 glioma cells 

Notes

Acknowledgments

Supported by the National Natural Science Foundation of China (No.30972470, 30771802), the grants from the National High Technology Research and Development Programme (863 Programme) of China (No.2010AA023003), and the Funding Project for Academic Human Resources Development in Institutions of Higher Learning Under the Jurisdiction of Beijing Municipality (PHR201006112).

Conflict of interest

The authors declared none conflicts of interest.

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Wei-wei Ma
    • 1
  • Cheng-cheng Hou
    • 1
  • Xin Zhou
    • 1
  • Huan-ling Yu
    • 1
  • Yuan-di Xi
    • 1
  • Juan Ding
    • 1
  • Xia Zhao
    • 1
  • Rong Xiao
    • 1
    Email author
  1. 1.School of Public Health and Family MedicineCapital Medical UniversityBeijingPeople’s Republic of China

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