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Published on: December 26, 2013
Quantitative proteomic analysis reveals that lipopolysaccharide induces mitogen-activated protein kinase-dependent
Kyunghee Byun1, Jin Young Kim, Enkhjargal Bayarsaikhan
1Center for Genomics and Proteomics, Lee Gil Ya Cancer and Diabetes Institute, Gachon University, Incheon, Korea.
Abstract:
Microglial cells act as the first and main form of active immune defense in the central nervous system related to inflammation and neurodegenerative disease. Lipopolysaccharide (LPS) induces many genes encoding inflammatory mediators, including cytokines such as tumor necrosis factor-α, interleukin-1β, (IL-1β), and IL-6, chemokines, and prostaglandins in microglial cells. Quantitative proteomics methods with isobaric chemical labeling using tandem mass tags and 2D-nano LC-ESI-MS/MS were used to systematically analyze proteomic changes in microglia responding to LPS stimulation. As a result, we found that the expression level of 21 proteins in human microglial cells changed after activation. Among those, one of the strong mitogen-activated protein kinase (MAPK) regulator proteins, CMPK1 was highly upregulated after LPS stimulation in human microglial cells. We detected and validated upregulation of MAPK including ERK1/2, p38, and SAPK/JNK by immunohistochemistry and Western blotting. NFκB, strong transcription factor of CMPK1, was translocated to the nucleus from the cytosol by high contents screening after LPS stimulation. Taken together, we conclude that MAPK signaling plays an important role in LPS-induced human microglial activation related to inflammatory response.
Insights
Lipopolysaccharide (LPS) activates human microglial cells, upregulating mitogen-activated protein kinase (MAPK) signaling pathways. This study identifies key protein changes, including CMPK1, crucial for inflammatory responses in the central nervous system.
Area of Science:
- Neuroscience
- Immunology
- Proteomics
Background:
- Microglial cells are key immune defenders in the central nervous system.
- Lipopolysaccharide (LPS) stimulation triggers inflammatory gene expression in microglia, relevant to neurodegenerative diseases.
Purpose of the Study:
- To systematically analyze proteomic changes in human microglial cells upon LPS activation.
- To identify key signaling pathways involved in LPS-induced microglial inflammatory responses.
Main Methods:
- Quantitative proteomics using tandem mass tags (TMT) and 2D-nano LC-ESI-MS/MS.
- Immunohistochemistry and Western blotting to validate protein expression.
- High-content screening to assess nuclear translocation of transcription factors.
Main Results:
- LPS stimulation altered the expression of 21 proteins in human microglial cells.
- CMPK1, a mitogen-activated protein kinase (MAPK) regulator, was significantly upregulated.
- MAPK pathways (ERK1/2, p38, SAPK/JNK) and NFκB activation were confirmed.
Conclusions:
- MAPK signaling is critically involved in LPS-induced human microglial activation.
- Understanding these pathways offers insights into neuroinflammation and neurodegenerative disease mechanisms.
