Ferulic acid prevents LPS-induced up-regulation of PDE4B and stimulates the cAMP/CREB signaling pathway in PC12 cells.
OBJECTIVE
Phosphodiesterase 4 (PDE4) isozymes are involved in different functions, depending on their patterns of distribution in the brain. The PDE4 subtypes are distributed in different inflammatory cells, and appear to be important regulators of inflammatory processes. In this study we examined the effects of ferulic acid (FA), a plant component with strong anti-oxidant and anti-inflammatory activities, on lipopolysaccharide (LPS)-induced up-regulation of phosphodiesterase 4B (PDE4B) in PC12 cells, which in turn regulated cellular cAMP levels and the cAMP/cAMP response element binding protein (CREB) pathway in the cells.


METHODS
PC12 cells were treated with LPS (1 μg/mL) for 8 h, and the changes of F-actin were detected using laser scanning confocal microscopy. The levels of pro-inflammatory cytokines were measured suing ELISA kits, and PDE4B-specific enzymatic activity was assessed with a PDE4B assay kit. The mRNA levels of PDE4B were analyzed with Q-PCR, and the protein levels of CREB and phosphorylated CREB (pCREB) were determined using immunoblotting. Furthermore, molecular docking was used to identify the interaction between PDE4B2 and FA.


RESULTS
Treatment of PC12 cells with LPS induced thick bundles of actin filaments appearing in the F-actin cytoskeleton, which were ameliorated by pretreatment with FA (10-40 μmol/L) or with a PDE4B inhibitor rolipram (30 μmol/L). Pretreatment with FA dose-dependently inhibited the LPS-induced production of TNF-α and IL-1β in PC12 cells. Furthermore, pretreatment with FA dose-dependently attenuated the LPS-induced up-regulation of PDE4 activity in PC12 cells. Moreover, pretreatment with FA decreased LPS-induced up-regulation of the PDE4B mRNA, and reversed LPS-induced down-regulation of CREB and pCREB in PC12 cells. The molecular docking results revealed electrostatic and hydrophobic interactions between FA and PDE4B2.


CONCLUSIONS
The beneficial effects of FA in PC12 cells might be conferred through inhibition of LPS-induced up-regulation of PDE4B and stimulation of cAMP/CREB signaling pathway. Therefore, FA may be a potential therapeutic intervention for the treatment of neuroinflammatory diseases such as AD.
College of Life Science and Bioengineering, Beijing University of Technology, Beijing 100124, China.|Beijing Institute of Radiation Medicine, Beijing 100850, China.
Hao
H
Huang
No 97 Hospital of CPLA, Xuzhou 221004, China.
Qian
Q
Hong
Beijing Institute of Radiation Medicine, Beijing 100850, China.
Hong-Ling
HL
Tan
Beijing Institute of Radiation Medicine, Beijing 100850, China.
Cheng-Rong
CR
Xiao
Beijing Institute of Radiation Medicine, Beijing 100850, China.
Yue
Y
Gao
D000818:Animals; D003373:Coumaric Acids / Q000494:pharmacology*; D000242:Cyclic AMP / Q000378:metabolism*; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4 / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D062105:Molecular Docking Simulation; D016716:PC12 Cells; D051381:Rats; D015398:Signal Transduction; D015854:Up-Regulation
D003373:Coumaric Acids; D017362:Cyclic AMP Response Element-Binding Protein; D008070:Lipopolysaccharides; C004999:ferulic acid; D000242:Cyclic AMP; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4; C546993:PDE4B protein, rat
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