Thrombin induces expression of cytokine-induced SH2 protein (CIS) in rat brain astrocytes: Involvement of phospholipase A2, cyclooxygenase, and lipoxygenase

• Published in: Glia 2004-11, Vol.48 (2), p.102-111
• Main Authors: Ji, Kyung-ae, Yang, Myung-Soon, Jou, Ilo, Shong, Min-Ho, Joe, Eun-Hye
• Format: Article
• Language: English
• Subjects: Animals; Animals, Newborn; Astrocytes - drug effects; Astrocytes - enzymology; Astrocytes - metabolism; Biological and medical sciences; Cells, Cultured; Dinoprostone - metabolism; Dinoprostone - pharmacology; DNA-Binding Proteins - drug effects; DNA-Binding Proteins - metabolism; Encephalitis - chemically induced; Encephalitis - metabolism; Enzyme Inhibitors - pharmacology; Feedback, Physiological - drug effects; Feedback, Physiological - genetics; Free Radical Scavengers - pharmacology; Fundamental and applied biological sciences. Psychology; Immediate-Early Proteins - drug effects; Immediate-Early Proteins - genetics; Immediate-Early Proteins - metabolism; inflammation; Inflammation Mediators - metabolism; Interferon-gamma - metabolism; Interferon-gamma - pharmacology; Isolated neuron and nerve. Neuroglia; Leukotriene B4 - metabolism; Leukotriene B4 - pharmacology; Lipoxygenase - drug effects; Lipoxygenase - metabolism; neuroimmunology; Phospholipases A - drug effects; Phospholipases A - metabolism; Phospholipases A2; Prostaglandin-Endoperoxide Synthases - drug effects; Prostaglandin-Endoperoxide Synthases - metabolism; Rats; Rats, Sprague-Dawley; RNA, Messenger - drug effects; RNA, Messenger - metabolism; signal transduction; STAT1 Transcription Factor; Suppressor of Cytokine Signaling Proteins; Thrombin - pharmacology; Trans-Activators - drug effects; Trans-Activators - metabolism; Up-Regulation - drug effects; Up-Regulation - genetics; Vertebrates: nervous system and sense organs
• ISSN: 0894-1491; 1098-1136
• DOI: 10.1002/glia.20059

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 A2 (cPLA2), cyclooxygenase (COX), and lipoxygenase (LO) play a role in CIS expression, since inhibitors of cPLA2, 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 E2 (PGE2) and leukotriene B4 (LTB4), products of COX and LO, respectively, potentiated thrombin‐induced CIS expression, indicating that ROS, and PGE2 and LTB4 generated by COX and LO, mediate CIS expression. Since interferon‐γ (IFN‐γ)‐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 cPLA2, COX and LO pathways results in CIS expression. More importantly, CIS expression may be a negative feedback mechanism that prevents prolonged inflammatory responses. © 2004 Wiley‐Liss, Inc.