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Involvement of Src-Suppressed C Kinase Substrate in Neuronal Death Caused by the Lipopolysaccharide-Induced Reactive Astrogliosis

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Abstract

Src-suppressed C kinase substrate (SSeCKS), a protein kinase C substrate, is a major lipopolysaccharide (LPS) response protein, regulating the inflammatory process. In the process of spinal inflammatory diseases by LPS intraspinal injection, expression of SSeCKS in the spinal cord was increased, mainly in active astrocytes and neurons. Induced SSeCKS was colabeled with terminal deoxynucleotidyl transferase-mediated biotinylated-dUTP nick-end labeling (an apoptosis maker) in the late inflammation processes. These results indicated that SSeCKS might correlate with the inflammatory reaction and late neurodegeneration after LPS injection. A cell type-specific action for SSeCKS was further studied within C6 cells and PC12 cells. Knockdown of SSeCKS by small-interfering RNAs (siRNAs) blocked the LPS-induced inducible nitric oxide synthase (iNOS) expression in C6 cells, while overexpression SSeCKS enhanced iNOS expression. SSeCKS is also participated in regulation of PC12 cell viability. Loss of SSeCKS rescued PC12 cell viability, and excessive SSeCKS exacerbated the cell death upon conditioned medium and tumor necrosis factor-alpha exposure. This study delineates that SSeCKS may be important for host defenses in spinal inflammation and suggests a valuable molecular mechanism by which astrocytes modify neuronal viability during pathological states.

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Abbreviations

CNS:

Central nervous system

SSeCKS:

Src-suppressed C kinase substrate

LPS:

Lipopolysaccharide

PKC:

Protein kinase C

TUNEL:

Terminal deoxynucleotidyl transferase-mediated biotinylated-dUTP nick-end labeling

CM:

Conditioned medium

TNF-α:

Tumor necrosis factor α

iNOS:

Induced nitrogen monoxidum

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

FBS:

Fetal bovine serum

HS:

Horse serum

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Nos. 30770488 and 30870320); Natural Science Foundation of Jiangsu province (Nos. BK2009156 and BK2009157); and Health Project of Jiangsu Province (H200632).

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Authors and Affiliations

  1. Department of Immunology, Medical College, Nantong University, Nantong, 226001, People’s Republic of China

    Ping Wang, Linlin Sun, Aiguo Shen, Haiou Liu, Tao Tao & Chun Cheng

  2. Laboratory Center, Affiliated Hospital of Nantong University, Nantong, 226001, People’s Republic of China

    Junling Yang

  3. The Jiangsu Province Key Laboratory of Neuroregeneration, Nantong University, Nantong, 226001, People’s Republic of China

    Xiaohong Li

  4. Department of Geriatrics, Nanjing Second Hospital affiliated to Nanjing Medical University, 121 Jiangjiayuan, Nanjing, 210011, People’s Republic of China

    Xiang Lu

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  1. Ping Wang
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Corresponding authors

Correspondence to Chun Cheng or Xiang Lu.

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Ping Wang and Linlin Sun contributed equally to this work.

Grant sponsor: National Natural Science Foundation of China (Nos. 30770488 and 30870320); Natural Science Foundation of Jiangsu province (Nos. BK2009156 and BK2009157); Health Project of Jiangsu Province (H200632)

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Wang, P., Sun, L., Shen, A. et al. Involvement of Src-Suppressed C Kinase Substrate in Neuronal Death Caused by the Lipopolysaccharide-Induced Reactive Astrogliosis. Inflammation 33, 359–373 (2010). https://doi.org/10.1007/s10753-010-9194-3

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