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Harvesting Murine Alveolar Macrophages and Evaluating Cellular Activation Induced by Polyanhydride Nanoparticles
Published on: June 8, 2012
Activation of RAW 264.7 cells by a polysaccharide isolated from Antarctic bacterium Pseudoaltermonas sp. S-5
Jing Li1, Wen Qian2, Yanghui Xu2
1Anhui Provincial Engineering Research Center for Polysaccharide Drugs, Anhui Province Key Laboratory of Active Biological Macro-Molecules, Department of Pharmacy, Wannan Medical College, Wuhu, 241000, China; Department of Pharmacy, Lu'an Affiliated Hospital of Anhui Medical University, Lu'an, 237000, China.
Abstract:
The aim of this study was to examine the effect of extracellular polysaccharide (PEP) from Antarctic bacterium Pseudoaltermonas sp. S-5 on RAW 264.7 cells together with the underlying signaling pathways. Our results illustrated that PEP induced dendritic-like morphological change in RAW 264.7 cells, and increased the productions of nitric oxide (NO) and tumor necrosis factor-α (TNF-α). PEP could also enhance phagocytic activity of RAW 264.7 cells. Results of immunofluorescence staining and immunoblotting indicated that PEP caused the nuclear translocation of nuclear factor (NF)-κB subunit p65, the degradation of IκB-α and up-expression of phosphorylated p38 mitogen-activated protein kinase (MAPK) in RAW 264.7 cells. According to pharmacological evaluation with specific enzyme inhibitors, both NF-κB and p38 MAPK signaling pathways were involved in the generation of NO and TNF-α induced by PEP. All these results indicated that PEP from Antarctic bacterium Pseudoaltermonas sp. S-5 activated RAW 264.7 cells through NF-κB and p38 MAPK signaling pathways.
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