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Protective Effects of ShcA Protein Silencing for Photothrombotic Cerebral Infarction

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Abstract

Reactive oxygen species (ROS) exacerbate stroke-induced cell damage. We found that ShcA, a protein that regulates ROS, is highly expressed in a Rose Bengal photothrombosis model. We investigated whether ShcA is essential for mitophagy in ROS-induced cellular damage and determined whether ROS exacerbate mitochondrial dysfunction via ShcA protein expression. Ischemic stroke was generated by Rose Bengal photothrombosis in mice. To silence ShcA protein expression in the mouse brain, ShcA-targeting siRNA-encapsulated nanoparticles were intrathecally injected into the cisterna magna. Upon staining with antibodies against ShcA counterpart caspase-3 or NeuN, we found that the ShcA protein expression was increased in apoptotic neurons. In addition, mitochondrial dysfunction and excessive mitophagy were evident in photothrombotic stroke tissue. Infarct volumes were significantly reduced, and neurological deficits were diminished in the ShcA siRNA nanoparticle-treated group, compared with the negative control siRNA nanoparticle-treated group. We confirmed that the reduction of ShcA expression by nanoparticle treatment rescued the expression of genes, associated with mitochondrial dynamics and mitophagy mediation in a stroke model. This study suggests that the regulation of ShcA protein expression can be a therapeutic target for reducing brain damage with mitochondrial dysfunction caused by thrombotic infarction.

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Funding

This research was supported by the Ministry of Science, ICT & Future Planning (NRF-2019R1C1C1005585, NRF-2018R1D1A1B07045664), and the Brain Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning (NRF-2019R1A2C2004884).

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Authors and Affiliations

  1. Department of Medical Science, Chungnam National University College of Medicine, Daejeon, 35015, Republic of Korea

    Jeong-Ah Hwang, Nara Shin, Hyo Jung Shin, Yuhua Yin, Hyeok Hee Kwon, Hyewon Park, Juhee Shin, Song I Kim & Dong Woon Kim

  2. Department of Anatomy and Cell Biology, Brain Research Institute, Chungnam National University College of Medicine, Daejeon, 35015, Republic of Korea

    Jeong-Ah Hwang, Nara Shin, Hyo Jung Shin, Yuhua Yin, Hyeok Hee Kwon, Hyewon Park, Juhee Shin, Song I Kim & Dong Woon Kim

  3. Brain Korea 21 PLUS Project for Medical Science, Chungnam National University College of Medicine, Daejeon, 35015, Republic of Korea

    Jeong-Ah Hwang

  4. Department of Neurology, Chungnam National University College of Medicine and Sejong Hospital, Sejong, 30099, Republic of Korea

    Hee-Jung Song

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  1. Jeong-Ah Hwang
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Contributions

JH participated in the design of the study, carried out pain behavior testing and immunostaining studies, participated in data analysis, and drafted the manuscript. NS and HJS carried out real-time PCR. YY and HHK participated in the RB model and behavior tests. HP and SK participated in in vitro experiments. DWK and HS participated in the design of the study and data analysis and wrote the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Dong Woon Kim or Hee-Jung Song.

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The authors declare that they have no conflict of interest.

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All protocols were performed in accordance with the Chungnam National University hospital Guide for the Care and Use of Laboratory Animals and were approved by the University of Chungnam Institutional Animal Care and Use Committee.

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Hwang, JA., Shin, N., Shin, H.J. et al. Protective Effects of ShcA Protein Silencing for Photothrombotic Cerebral Infarction. Transl. Stroke Res. 12, 866–878 (2021). https://doi.org/10.1007/s12975-020-00874-1

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  • DOI: https://doi.org/10.1007/s12975-020-00874-1

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