Comparison of prostaglandin F2alpha, bimatoprost (prostamide), and butaprost (EP2 agonist) on Cyr61 and connective tissue growth factor gene expression.
Connective tissue growth factor (CTGF) and Cyr61 (cysteine-rich angiogenic protein 61) are members of the CCN gene family that encode multifunctional, extracellular matrix-associated signaling proteins. Because the mechanism of action of certain anti-glaucoma drugs involves extracellular matrix remodeling of ocular ciliary muscle, with a resultant increase in drainage of aqueous humor from the eye, we compared the effects of three pharmacologically distinct ocular hypotensive agents on Cyr61 and CTGF gene expression. Thus, prostaglandin F2alpha (PGF2alpha) (FP receptor agonist), Butaprost (EP2 receptor agonist), and Bimatoprost (a prostamide) were compared. Using Affymetrix gene chip technology, we first identified that PGF2alpha dramatically up-regulated Cyr61 and CTGF mRNA expression in HEK 293/EBNA cells (hFP-HEK 293/EBNA). Northern blot further confirmed the Cyr61 and CTGF up-regulation is in a dose- and time-dependent manner. PGF2alpha-induced up-regulation of Cyr61 appeared to exclusively involve the Rho pathway, and up-regulation of CTGF was via multiple intracellular pathways. Because prostamide receptors are, to date, defined only at the pharmacological level, Bimatoprost effects on Cyr61 and CTGF were studied in the isolated feline iris sphincter preparation, a tissue highly responsive to prostamides. Both PGF2alpha and Bimatoprost up-regulated Cyr61 mRNA expression in the cat iris tissue. Only PGF2alpha up-regulated CTGF mRNA expression in the cat iris. Therefore, PGF2alpha and Bimatoprost appear to interact with different receptors populations in the cat iris, according to their markedly different effects on CTGF. Activation of prostaglandin EP2 receptors (Gs-coupled) also up-regulated Cyr61 but not CTGF mRNA expression in the isolated cat iris. Similar data were observed in human primary ciliary smooth muscle cells. Thus, despite quite different signal transduction pathways, FP receptor stimulation up-regulates CTGF and Cyr61. The prostamide analog Bimatoprost and an EP2-selective agonist affects only Cyr61.
Allergan, Inc., Irvine, California 92612, USA.
Yanbin
Y
Liang
Chen
C
Li
Victor M
VM
Guzman
Albert J
AJ
Evinger
Charles E
CE
Protzman
Achim H-P
AH
Krauss
David F
DF
Woodward
D000527:Alprostadil / Q000031:analogs & derivatives* / Q000494:pharmacology*; D000577:Amides; D000818:Animals; D000069580:Bimatoprost; D002415:Cats; D002460:Cell Line; D002478:Cells, Cultured; D002924:Ciliary Body / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D003008:Cloprostenol / Q000031:analogs & derivatives; D055513:Connective Tissue Growth Factor; D055515:Cysteine-Rich Protein 61; D015237:Dinoprost / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D005901:Glaucoma / Q000188:drug therapy; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics*; D066298:In Vitro Techniques; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics*; D007498:Iris / Q000187:drug effects / Q000378:metabolism; D007700:Kinetics; D008055:Lipids / Q000494:pharmacology*; D020411:Oligonucleotide Array Sequence Analysis; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014129:Trabecular Meshwork / Q000187:drug effects / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects
D000577:Amides; C501848:CCN1 protein, human; C526317:CCN2 protein, human; D055515:Cysteine-Rich Protein 61; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D008055:Lipids; D012333:RNA, Messenger; D055513:Connective Tissue Growth Factor; D003008:Cloprostenol; D015237:Dinoprost; D000527:Alprostadil; C048491:butaprost; D000069580:Bimatoprost