constitutive expression of functionally active protease-activated receptors 1 and 2 in human conjunctival epithelial cells组成型表达功能活跃的protease-activated人类结膜上皮细胞受体1和2.pdfVIP

Hindawi Publishing Corporation Mediators of Inflammation Volume 2006, Article ID 61359, Pages 1–8 DOI 10.1155/MI/2006/61359 Research Communication Constitutive Expression of Functionally Active Protease-Activated Receptors 1 and 2 in Human Conjunctival Epithelial Cells Timothy J. Nickel, Mohammad H. Kabir, Jaya Talreja, Daniel J. Stechschulte, and Kottarappat N. Dileepan Division of Allergy, Clinical Immunology, and Rheumatology, Department of Medicine, The University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, KS 66160, USA Received 17 February 2006; Accepted 1 March 2006 Protease-activated receptors (PARs) are G-protein-coupled receptors which initiate inflammatory responses when activated by specific serine proteases. This study was conducted to examine whether human conjunctival epithelial cells (HCECs) express functionally active PAR1 and PAR2 using Chang conjunctival epithelial cells as in vitro model. We performed RT-PCR and im- munofluorescence analyses to determine the expression of PAR1 and PAR2, and monitored the production of IL-6 after activating HCECs with PAR1 activating agents (thrombin or TFLLRN) or PAR2 activating agents (tryptase, trypsin, or SLIGKV). The results show that HCECs constitutively express PAR1 and PAR2 mRNA and proteins, and produce significant amounts of IL-6 when in- cubated with specific PAR-activating enzymes or agonist peptides. Thrombin- and tryptase-induced HCEC activation was blocked by PAR1 and PAR2 neutralizing antibodies, respectively, and by specific enzyme inhibitors. The constitutive expression of PAR1 and PAR2, and their activation by thrombin and tryptase, respectively, may have important implications in ocular inflammation. Copyright © 2006 Timothy J. Nickel et al. This is an open access article distributed under the Creative Comm