The regulation of pro-inflammatory gene expression induced by pigment epithelium-derived factor in rat cultured microglial cells.
Pigment epithelium-derived factor (PEDF) is a potent and broadly acting neurotrophic factor that protects various cultured neurons against apoptotic stimuli. To investigate whether PEDF acts not only on neurons, but also glial cells, we analyzed the effects of recombinant human PEDF (rhPEDF) on cytokine mRNA levels, transcription factors, and signal transduction pathways in cultured microglial cells. RT-PCR analysis revealed that pro-inflammatory genes such as IL-1beta, IL-6, and TNFalpha were induced in rhPEDF-treated cultured microglial cells. Furthermore, rapid phosphorylation of CREB protein had occurred in rhPEDF-treated neonatal astrocytes. Up-regulation of pro-inflammatory genes by rhPEDF was blocked by overexpression of dominant negative CREB or a mutated form of IkappaBalpha. These results suggest that the induction of pro-inflammatory genes by rhPEDF is mediated via activation of NF-kappaB or CREB in microglial cells. Taken together, these results demonstrate that PEDF is a multipotent factor, capable of affecting not only neurons, but also microglial cells, and suggests that it may act as a neuro-immune modulator in the rodent brain.
Sanagi
Tomomi
T
Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8641, Japan.
Yabe
Takeshi
T
Yamada
Haruki
H
D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism; D021983:Cyclophilins / Q000235:genetics / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005136:Eye Proteins / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D007249:Inflammation / Q000235:genetics* / Q000378:metabolism; D007976:Leupeptins / Q000494:pharmacology; D017628:Microglia / Q000187:drug effects*; D048052:Mitogen-Activated Protein Kinase 3 / Q000235:genetics / Q000378:metabolism; D009414:Nerve Growth Factors / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D015843:Serpins / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D000099174:Pigment Epithelium-Derived Factor
D017362:Cyclic AMP Response Element-Binding Protein; D021983:Cyclophilins; D015853:Cysteine Proteinase Inhibitors; D016207:Cytokines; D005136:Eye Proteins; D007976:Leupeptins; D048052:Mitogen-Activated Protein Kinase 3; D009414:Nerve Growth Factors; D012333:RNA, Messenger; D011994:Recombinant Proteins; D015843:Serpins; D014409:Tumor Necrosis Factor-alpha; D048051:p38 Mitogen-Activated Protein Kinases; D000099174:Pigment Epithelium-Derived Factor; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde