Thrombin induces expression of cytokine-induced SH2 protein (CIS) in rat brain astrocytes: involvement of phospholipase A2, cyclooxygenase, and lipoxygenase.
Previously we have reported that thrombin induces inflammatory mediators in brain glial cells (Ryu et al. 2000. J Biol Chem 275:29955). In the present study, we found that thrombin induced a negative regulator of a cytokine signaling molecule, cytokine-induced SH2 protein (CIS), in rat brain astrocytes. In response to thrombin, CIS expression was increased at both the mRNA and protein levels. Although STAT5 is known to regulate CIS expression, thrombin did not activate STAT5, and inhibitors of JAK2 (AG490) and JAK3 (WHI-P97 and WHI-P154) had little effect on thrombin-induced CIS expression. In contrast, cytosolic phospholipase A(2) (cPLA(2)), cyclooxygenase (COX), and lipoxygenase (LO) play a role in CIS expression, since inhibitors of cPLA(2), cyclooxygenase (COX), and LO significantly reduced CIS expression. Reactive oxygen species (ROS) scavengers (N-acetyl-cysteine [NAC] and trolox) reduced thrombin-induced CIS expression, and inhibitors of COX and LO reduced ROS produced by thrombin. Furthermore, prostaglandin E(2) (PGE(2)) and leukotriene B(4) (LTB(4)), products of COX and LO, respectively, potentiated thrombin-induced CIS expression, indicating that ROS, and PGE(2) and LTB(4) generated by COX and LO, mediate CIS expression. Since interferon-gamma (IFN-gamma)-induced GAS-luciferase activity and tyrosine phosphorylation of STAT1 and STAT3 were lower in CIS-transfected cells compared to control vector-transfected cells, CIS could have anti-inflammatory activity. These data suggest that thrombin-stimulation of ROS and prostaglandin and leukotriene production via the cPLA(2), COX and LO pathways results in CIS expression. More importantly, CIS expression may be a negative feedback mechanism that prevents prolonged inflammatory responses.
Ji
Kyung-Ae
KA
Neuroscience Graduate Program, Brain Disease Research Center, Ajou University School of Medicine, Suwon, Korea.
Yang
Myung-Soon
MS
Jou
Ilo
I
Shong
Min-Ho
MH
Joe
Eun-Hye
EH
D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000187:drug effects / Q000378:metabolism; D015232:Dinoprostone / Q000378:metabolism / Q000494:pharmacology; D004660:Encephalitis / Q000139:chemically induced / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D025461:Feedback, Physiological / Q000187:drug effects / Q000235:genetics; D016166:Free Radical Scavengers / Q000494:pharmacology; D017874:Immediate-Early Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D018836:Inflammation Mediators / Q000378:metabolism*; D007371:Interferon-gamma / Q000378:metabolism / Q000494:pharmacology; D007975:Leukotriene B4 / Q000378:metabolism / Q000494:pharmacology; D008084:Lipoxygenase / Q000187:drug effects / Q000378:metabolism; D010741:Phospholipases A / Q000187:drug effects / Q000378:metabolism; D054467:Phospholipases A2; D011451:Prostaglandin-Endoperoxide Synthases / Q000187:drug effects / Q000378:metabolism; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D050794:STAT1 Transcription Factor; D050826:Suppressor of Cytokine Signaling Proteins; D013917:Thrombin / Q000494:pharmacology*; D015534:Trans-Activators / Q000187:drug effects / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics
D004268:DNA-Binding Proteins; D015232:Dinoprostone; D004791:Enzyme Inhibitors; D016166:Free Radical Scavengers; D017874:Immediate-Early Proteins; D018836:Inflammation Mediators; D007371:Interferon-gamma; D007975:Leukotriene B4; D008084:Lipoxygenase; D010741:Phospholipases A; D054467:Phospholipases A2; D011451:Prostaglandin-Endoperoxide Synthases; D012333:RNA, Messenger; D050794:STAT1 Transcription Factor; D050826:Suppressor of Cytokine Signaling Proteins; D013917:Thrombin; D015534:Trans-Activators; C494081:Stat1 protein, rat