JOURNAL OF OCULAR PHARMACOLOGY AND THERAPEUTICS
`Volume 24, Number 4, 2008
`© Mary Ann Liebert, Inc.
`DOI: 10.1089/jop.2008.0042
`
`Latanoprost-Induced Changes in Rat Intraocular Pressure:
`Direct or Indirect?
`
`Shahid Husain, Phillip W. Yates, and Craig E. Crosson
`
`Abstract
`
`Introduction: The topical application of prostaglandin F2␣ (FP)-receptor agonists has been shown to signifi-
`cantly lower intraocular pressure (IOP) in humans and is now considered the first-line treatment for ocular hy-
`pertension. Despite the prominent role FP-receptor agonists play in the treatment of glaucoma, our under-
`standing of how these agents lower IOP remains incomplete. The present study was designed to evaluate the
`role of matrix metalloproteinase (MMP) activation and the cytokine, tumor necrosis factor alpha (TNF-␣), in
`latanoprost-induced changes in IOP.
`Methods: Changes in IOP following an acute topical administration of latanoprost (60 ng) in normotensive
`Brown Norway rats were evaluated by means of a commercially available rebound tonometer. To examine the
`role of MMPs and TNF-␣ in this response, the rats were pretreated with a broad-spectrum MMP inhibitor, GM-
`6001 (100 ␮g), or the TNF-␣ inhibitor, thalidomide (25 ␮g).
`Results: The topical administration of latanoprost (60 ng) alone produced a biphasic change in ipsilateral IOP:
`an initial hypertension (4.21 ⫾ 0.52), followed by a prolonged hypotension (⫺4.79 ⫾ 0.65). In rats, pretreatment
`with GM-6001 blocked the latanoprost-induced reduction in IOP but did not prevent the initial rise in IOP. Pre-
`treatment with thalidomide also blocked the ocular hypotension induced by latanoprost; however, thalidomide
`pretreatment enhanced the duration of the initial hypertension.
`Conclusions: These results provide evidence that the secretion and activation of MMPs and the release of TNF-
`␣ play a central role in the ocular hypotension induced by FP-agonists. The administration of FP-agonists ap-
`pears to lower IOP directly by inducing the activation of MMPs within the ciliary body, leading to improved
`uveoscleral outflow and indirectly through the release of TNF-␣ within the ciliary body. Secreted TNF-␣ may
`then activate TNF-receptors in the uvea and trabecular meshwork, increasing both uveoscleral and conven-
`tional outflow.
`
`Introduction
`
`THE PRIMARY RISK FACTOR FOR the development of glaucoma
`
`is elevated intraocular pressure (IOP). Topical applica-
`tion of prostaglandin F2␣ (FP) or its analogs have been shown
`to significantly reduce IOP in humans1,2 and primates.3,4
`However, early animal studies demonstrated that this re-
`sponse is unusual, in that the response to FP-receptor ago-
`nists improves with multiple dosing over time.5–7 This en-
`hanced functional response supports the idea that the
`chronic use of FP-receptor agonists induces a structural
`and/or molecular change within the outflow pathway that
`acts synergistically with a later administration of an FP-re-
`ceptor agonist to lower IOP.
`
`Topical treatment with prostaglandin F2␣ analogs alters
`the extracellular matrix (ECM) in the tissues of the uveoscle-
`ral outflow pathway8 and increases the production of ma-
`trix metalloproteinases (MMPs) in the ciliary muscle cells.8–15
`Although studies have shown that MMPs can alter conven-
`tional outflow facility in human and animal perfused
`eyes,16,17 direct evidence that MMPs participate in the FP-
`agonist-induced modulation of IOP remains to be deter-
`mined.
`In other systems, the activation of FP-receptors has been
`shown to induce the expression of the cytokines.18–20 In the
`eye, tumor necrosis factor alpha (TNF-␣) induces the secre-
`tion of MMPs and ECM remodeling in a number of tissues,
`including the trabecular meshwork, cornea, and optic nerve
`
`Hewitt Laboratory of the Ola B. Williams Glaucoma Center, Department of Ophthalmology, Medical University of South Carolina,
`Charleston, SC.
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`HUSAIN ET AL.
`
`head.21–25 Although TNF-␣ has been shown to increase con-
`ventional outflow facility, the participation of this cytokine
`in modulating uveoscleral outflow has not been investigated.
`To evaluate the roles of TNF-␣ and MMPs in the la-
`tanoprost-induced changes in IOP, Brown Norway rats were
`pretreated with an inhibitor of TNF-␣ production, thalido-
`mide, or the MMP inhibitor, GM-6001. Rats were selected for
`this study because they share similar anatomic and devel-
`opmental characteristics in the anterior segment with those
`of the human.26–29 Therefore, aqueous-humor-dynamic stud-
`ies in rats may be more predictive of the response in humans
`than those from other nonprimate laboratory animals. Re-
`sults from these studies support the idea that both TNF-␣
`and MMPs play a role in prostaglandin-induced changes in
`IOP.
`
`Methods
`
`Animals
`
`Normotensive, male Brown Norway (200–300 g) rats were
`used in this study. All animals were housed under standard
`laboratory conditions of a 12-h light and 12-h dark cycle. An-
`imal handling was in accordance with the Association for
`Research in Vision and Ophthalmology Statement for the
`Use of Animals in Ophthalmic and Vision Research, and
`study protocols were approved by the Animal Care and Use
`Committee at the Medical University of South Carolina
`(Charleston, SC).
`
`Drug treatment
`
`Latanoprost (0.005%; Pfizer, San Diego, CA) was diluted
`in saline to a final concentration of 0.002% and administered
`topically (3 ␮L). Thalidomide (0.25%; Sigma Chemical Com-
`pany, St Louis, MO) or GM-6001 (0.05%; Calbiochem, La
`Jolla, CA) was formulated in saline containing 10% dimethyl
`
`FIG. 1. Effect of latanoprost-induced intraocular pressure
`changes in Brown Norway rats. Rats were treated topically
`with 60 ng of latanoprost or vehicle (t ⫽ 0 h). Values are
`recorded as the mean ⫾ standard error of the mean. Aster-
`isks denote a significant (P ⬍ 0.05) difference between eyes
`treated with latanoprost alone and eyes receiving vehicle
`(n ⫽ 6).
`
`FIG. 2. Effect of the matrix metalloproteinase inhibitor,
`GM-6001, on
`latanoprost-induced
`intraocular pressure
`changes in Brown Norway rats. Rats were treated topically
`with: GM-6001 (100 ␮g) bilaterally 1 h prior to the topical,
`unilateral administration of 60 ng of latanoprost (t ⫽ 0 h).
`Values are recorded as the mean ⫾ standard error of the
`mean. Asterisks denote a significant (P ⬍ 0.05) difference be-
`tween eyes treated with latanoprost alone and eyes receiv-
`ing both GM-6001 and latanoprost (n ⫽ 6).
`
`sulfoxide (DMSO). For thalidomide pretreatment, the agent
`was topically administered as a 5 ␮L-drop, twice, 15 min
`apart. For GM-6001 pretreatment, the agent was topically ad-
`ministered as a 10-␮L drop, twice, 15 min apart.
`
`IOP measurement in rats
`
`IOP was measured in conscious rats by using a calibrated
`TonoLab (Colonial Medical Supply Co. Inc., Franconia, NH)
`rebound tonometer, as previously described.30 To minimize
`any discomfort to the animals during tonometry, corneas
`were lightly anesthetized by the topical application of 0.1%
`proparacaine (3 ␮L). Baseline IOPs of both eyes were deter-
`mined (t ⫽ ⫺1 h). Rats were then pretreated bilaterally with
`GM-6001, thalidomide, or vehicle. After 1 h, IOPs were again
`determined (t ⫽ 0 h) and latanoprost applied topically to 1
`eye. Ipsilateral and contralateral IOPs were then measured
`at 1, 2, 3, 4, 5, 6, and 24 h postlatanoprost treatment. All IOP
`measurements were recorded in a masked fashion. In sepa-
`rate studies, unilateral treatment with GM-6001 or thalido-
`mide alone was evaluated to determine if these agents pro-
`duced any acute change in IOP.
`
`Data analysis
`
`Values are presented as the mean ⫾ standard error of the
`mean. Ipsilateral IOPs from latanoprost-treated animals
`were compared with corresponding IOPs from animals pre-
`treated with thalidomide or GM-6001 by means of the Stu-
`dent t-test for nonpaired data. A P-value of ⱕ0.05 was con-
`sidered significant.
`
`Results
`
`In conscious Brown Norway rats, the mean IOP measured
`in this study was 18.6 ⫾ 1.2 mmHg. The topical administra-
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`LATANOPROST AND IOP
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`mmHg) at 5 h (Fig. 1). No significant change in contralateral
`IOP was noted in these animals.
`The administration of GM-6001 (100 ␮g) alone did not sig-
`nificantly alter IOP (Fig. 2). In eyes pretreated with GM-6001,
`the administration of latanoprost induced an initial increase
`in pressure at 1 h. However, the latanaprost-induced hyper-
`tension at 2 h and the subsequent hypotension from 4 to 6 h
`were completely blocked by pretreatment with GM-6001.
`The administration of thalidomide (25 ␮g) alone did not
`induce any significant change in IOP (Fig. 3). In eyes pre-
`treated with thalidomide, the duration of the early hyper-
`tensive response was enhanced, with pressures remaining
`elevated above baseline for up to 4 h. At times beyond 4 h,
`the ocular hypotensive response was significantly inhibited,
`when compared to eyes treated with latanoprost alone.
`No changes in conjunctival hyperemia or abnormal ani-
`mal behavior were observed during the course of these ex-
`periments.
`
`Discussion
`
`Topically administered prostaglandin F2␣ and its analogs
`lower IOP in human and nonhuman primates.1,3,4,31,32 Al-
`though these drugs are now considered first-line therapies
`for the treatment of glaucoma, the cellular mechanisms that
`mediate this response are poorly understood. The ocular hy-
`potensive response to FP-agonists is unusual, in that it im-
`proves over time with multiple dosing.5–7 This enhanced re-
`sponse supports the idea that the chronic use of FP-agonists
`induces a structural and/or molecular change within the out-
`flow pathway that acts synergistically with the subsequent
`administration of FP-agonists to lower IOP. Morphologic,
`molecular, and biochemical studies have demonstrated that
`
`FIG. 3. Effect of the tumor necrosis factor alpha inhibitor,
`thalidomide, on
`latanoprost-induced
`intraocular pressure
`changes in Brown Norway rats. Rats were treated bilaterally with
`thalidomide (25 ␮g) 1 h prior to the unilateral administration of
`60 ng of latanoprost (t ⫽ 0 h). Values are recorded as the mean ⫾
`standard error of the mean. Asterisks denote a significant (P ⬍
`0.05) difference between eyes treated with latanoprost alone and
`eyes receiving both thalidomide and latanoprost (n ⫽ 8).
`
`tion of 60 ng of latanoprost alone produced a biphasic change
`in IOP: an initial rise in pressure peaking at 1 h (4.21 ⫾ 0.52
`mmHg), followed by a prolonged hypotensive response
`from 4–6 h, with a maximum reduction in IOP (⫺4.79 ⫾ 0.65
`
`FIG. 4. Putative model for the ocular mechanisms responsible for intraocular pressure lowering by prostaglandin F2␣ (FP)-
`receptor agonists. Solid arrows show direct FP actions, whereas gray, dashed arrows indicate indirect actions.
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`HUSAIN ET AL.
`
`FP-agonists lead to a loss of connective tissue within the cil-
`iary body and increase the expression and/or secretion of
`MMPs.8,13,15,33 However, studies have not provided direct
`evidence that MMPs are involved in the ocular hypotensive
`response to FP-agonists.
`In vitro studies have shown that exposure to prostaglan-
`dins induces the secretion of MMPs from human ciliary mus-
`cle cells.9,11 In addition, a reduction in collagen types I, III,
`and IV in PGF2␣-treated human ciliary muscle cultures have
`been reported.14 Further, a reduction in these collagens was
`observed in the ciliary muscle of inflamed monkey eyes pre-
`viously treated with PGF2␣-IE, in which uveoscleral outflow
`was also increased.8,34 Thus, increased degradation of ciliary
`muscle ECM by MMP is thought to decrease the hydraulic
`resistance to uveoscleral outflow, thereby lowering the IOP
`in the eye. To provide direct evidence that the secretion and
`activation of MMPs was involved in the IOP responses to la-
`tanoprost, rats were pretreated with the broad-spectrum
`MMP inhibitor, GM-6001. GM-6001 is a hydroxamic acid de-
`rivative originally synthesized as an inhibitor of human skin
`collagenase35 and has been shown also to block MMP-1, -2,
`-3, and -9 activities.36,37 Pretreatment with GM-6001 blocked
`the ocular hypotensive responses induced by latanoprost.
`These results provide evidence, for the first time, that the se-
`cretion and activation of MMPs are required steps in the la-
`tanoprost-induced reduction in rat IOP.
`The initial hypertensive response seen at 1 h following a
`latanoprost administration was not altered. These results
`support the idea that the early hypertensive response is in-
`dependent of MMP secretion and activation within the an-
`terior segment of the eye. This response is likely mediated
`by the vasodilation of uveal vessels producing a rapid in-
`crease in uveal volume and IOP.
`TNF-␣ is a proinflammatory cytokine produced by a vari-
`ety of cells, including lymphocytes, astrocytes, and smooth
`muscle cells. In other systems, the activation of FP-receptors
`has been shown to induce the expression of the cytokine TNF-
`␣.18–20 In the anterior segment, elevation in TNF-␣ is thought
`to contribute to the increase in conventional outflow facility
`produced by argon laser trabeculoplasty and the breakdown
`of the blood-aqueous barrier.38,39 Thalidomide is a potent in-
`hibitor of TNF-␣ production.40,41 This inhibition is due to the
`increased degradation of TNF-␣ mRNA and reduced expres-
`sion by stabilizing NF␬B within the cytosol.42 However, it
`should be noted that thalidomide can also suppress the lev-
`els of interleukin (IL)-1␤, IL-6, and granulocyte macrophage-
`colony stimulating factor and enhance the levels of IL-10.43 In
`the current study, pretreatment with thalidomide suppressed
`the ocular hypotension induced by the addition of latanoprost.
`These data support the idea that the secretion of TNF-␣, or
`other cytokines, is, in part, responsible for the ocular hy-
`potensive response to FP-agonists. This release of TNF-␣
`might also provide an explanation for observations that FP-
`agonists can also increase conventional outflow facility.44,45
`Although FP-receptor expression is only a minor fraction of
`the total prostanoid-receptor expression in the trabecular
`meshwork,46 the latanoprost-induced secretion of TNF-␣ from
`the ciliary muscle may act in a paracrine fashion to stimulate
`MMP secretion from the trabecular meshwork, thus increas-
`ing conventional outflow indirectly.
`Rats pretreated with thalidomide also exhibited an en-
`hanced early hypertensive response. These data indicate that
`
`the full expression of the ocular hypertension induced by FP-
`agonists is masked by the hypotensive action of TNF-␣ re-
`leased into the intraocular environment.
`
`Conclusions
`In summary, we have shown that the topical application of
`latanoprost in rats produced a biphasic change in IOP: an ini-
`tial rise in pressure, followed by a prolonged hypotensive re-
`sponse. The prolonged hypotension could be blocked by pre-
`treatment with the general MMP inhibitor, GM-6001, or the
`TNF-␣ inhibitor, thalidomide. However, neither agent sup-
`pressed the initial hypertension induced in these animals.
`Taken together, these results provide evidence that the secre-
`tion and activation of MMP and the release of TNF-␣ play a
`role in the ocular hypotension induced by FP-agonists. Based
`on the results from this study and others, Figure 4 provides a
`putative working model for the ocular hypotensive actions of
`FP-agonists. In this model, FP-agonists lower IOP directly by
`inducing the activation of MMPs within the ciliary body, lead-
`ing to improved uveoscleral outflow. They also lower IOP in-
`directly through the release of TNF-␣ within the ciliary body.
`Secreted TNF-␣ may then activate TNF-receptors in the ciliary
`body, and possibly, the trabecular meshwork to increase both
`conventional and uveoscleral outflow.
`
`Acknowledgments
`This study was supported, in part, by NIH/NEI grants
`EY-09741 (C.E.C.) and EY-14793 (C.E.C.); the American
`Health Assistance Foundation (S.H.), NIH grant C-06
`RR015455 from the Extramural Research Facilities Program
`of the National Center for Research Resources, and an un-
`restricted grant to the Storm Eye Institute from Research to
`Prevent Blindness (New York, NY). The authors acknowl-
`edge the critical review of the original manuscript by L.
`Bartholomew, Ph.D.
`
`Disclosure
`The authors have no commercial relationships in the prod-
`ucts or equipment mentioned in this paper.
`
`References
`1. Camras, CB. Comparison of latanoprost and timolol in pa-
`tients with ocular hypertension and glaucoma: A six-month
`masked, multicenter trial in the United States. The United
`States Latanoprost Study Group. Ophthalmology 103:138–147,
`1996.
`2. Alm, A. Comparative phase III clinical trial of latanoprost
`and timolol in patients with elevated intraocular pressure.
`Adv. Prostagland. Thrombox. Leukot. Res. 23:527–532, 1995.
`3. Gabelt, B.T., and Kaufman, P.L. The effect of prostaglandin
`F2alpha on trabecular outflow facility in cynomolgus mon-
`keys. Exp. Eye Res. 51:87–91, 1990.
`4. Camras, C.B., and Bito, L.Z. Reduction of intraocular pres-
`sure in normal and glaucomatous primate (Aotus trivirgatus)
`eyes by topically applied prostaglandin F2alpha. Curr. Eye
`Res. 1:205–209, 1981.
`5. Wang, R.F., Camras, C.B., Lee, P.Y., et al. Effects of prosta-
`glandins F2alpha, A2, and their esters in glaucomatous mon-
`key eyes. Invest. Ophthalmol. Vis. Sci. 31:2466–2470, 1990.
`6. Crawford, K.S., and Kaufman, P.L. Dose-related effects of
`prostaglandin F2alpha isopropylester on intraocular pres-
`
`Micro Labs Exhibit 1046-4
`
`

`

`LATANOPROST AND IOP
`
`371
`
`sure, refraction, and pupil diameter in monkeys. Invest. Oph-
`thalmol. Vis. Sci. 32:510–519, 1991.
`7. Gabelt, B.T., Seeman, J.L., Podos, S.M., et al. Aqueous hu-
`mor dynamics in monkeys after topical 8-iso PGE(2). Invest.
`Ophthalmol. Vis. Sci. 45:892–899, 2004.
`8. Sagara, T., Gaton, D.D., Lindsey, J.D., et al. Topical prosta-
`glandin F2alpha treatment reduces collagen types I, III, and
`IV in the monkey uveoscleral outflow pathway. Arch. Oph-
`thalmol. 117:794–801, 1999.
`9. Husain, S., Jafri, F., and Crosson, C.E. Acute effects of
`PGF2alpha on MMP-2 secretion from human ciliary muscle
`cells, a PKC- and ERK-dependent process. Invest. Ophthal-
`mol. Vis. Sci. 46:1706–1713, 2005.
`10. Ocklind, A. Effect of latanoprost on the extracellular matrix
`of the ciliary muscle. A study on cultured cells and tissue
`sections. Exp. Eye Res. 67:179–191, 1998.
`11. Lindsey, J.D., Kashiwagi, K., Boyle, D., et al. Prostaglandins
`increase pro-MMP-1 and pro-MMP-3 secretion by human
`ciliary smooth muscle cells. Curr. Eye Res. 15:869–875, 1996.
`12. Weinreb, R.N., Kashiwagi, K., Kashiwagi, F., et al. Prosta-
`glandins increase matrix metalloproteinase release from hu-
`man ciliary smooth muscle cells. Invest. Ophthalmol. Vis. Sci.
`38:2772–2780, 1997.
`13. Gaton, D.D., Sagara, T., Lindsey, J.D., et al. Increased matric
`metalloproteinases 1, 2, and 3 in the monkey uveoscleral out-
`flow pathway after topical prostaglandin F2alpha-isopropyl
`ester treatment. Arch. Ophthalmol. 119:1165–1170, 2001.
`14. Lindsey, J.D., Kashiwagi, K., Kashiwagi, F., et al. Prosta-
`glandin action on ciliary smooth muscle extracellular matrix
`metabolism: Implications for uveoscleral outflow. Surv. Oph-
`thalmol. 41(Suppl 2):S53–S59, 1997.
`15. Lutjen-Drecoll, E., and Tamm, E. Morphological study of the
`anterior segment of cynomolgus monkey eyes following
`treatment with prostaglandin F2alpha. Exp. Eye Res.
`47:761–769, 1988.
`16. Crosson, C.E., Sloan, C.F., and Yates, P.W. Modulation of
`conventional outflow facility by the adenosine A1 agonist
`N6-cyclohexyladenosine.
`Invest. Ophthalmol. Vis. Sci.
`46:3795–3799, 2005.
`17. Bradley, J.M., Vranka, J., Colvis, C.M., et al. Effect of matrix
`metalloproteinases activity on outflow in perfused human
`organ culture. Invest. Ophthalmol. Vis. Sci. 39:2649–2658,
`1998.
`18. Fujino, H., and Regan, J.W. Prostaglandin F2alpha amplifies
`tumor necrosis factor-alpha promoter activity by the FPB
`prostanoid receptor. Biochem. Biophys. Res. Commun.
`317:1114–1120, 2004.
`19. Timmermann, M., and Hogger, P. Oxidative stress and 8-
`iso-prostaglandin F2a induce ectodomain shedding of CD163
`and release of tumor necrosis factor-alpha from human
`monocytes. Free Radic. Biol. Med. 39:98–107, 2005.
`20. Noguchi, K., Endo, H., Kondo, H., et al. Prostaglandin F2a
`upregulates interleukin-6 production in human gingival fi-
`broblasts. J. Endodontal. Res. 36:80–87, 2001.
`21. Alexander, J.P., Samples, J.R., and Acott, T.S. Growth factor
`and cytokine modulation of trabecular meshwork matrix
`metalloproteinase and TIMP expression. Curr. Eye Res.
`17:276–285, 1998.
`22. Samples, J.R., Alexander, J.P., and Acott, T.S. Regulation of
`the levels of human trabecular matrix metalloproteinases
`and inhibitor by interleukin-1 and dexamethasone. Invest.
`Ophthalmol. Vis. Sci. 34:3386–3395, 1993.
`23. Li, D.Q., Lokeshwar, B.L., Solomon, A., et al. Regulation of
`MMP-9 production by human corneal epithelial cells. Exp.
`Eye Res. 73:449–459, 2001.
`
`24. Yan, X., Tezel, G., Wax, M.B., et al. Matrix metalloproteinases
`and tumor necrosis factor alpha in glaucomatous optic nerve
`head. Arch. Ophthalmol. 118:666–673, 2000.
`25. Yuan, L., and Neufeld, A.H. Activated microglia in the hu-
`man glaucomatous optic nerve head. J. Neurosci. Res. 64:
`523–532, 2001.
`26. Daimon, T., Kazama, M., Miyajima, Y., et al. Immunocyto-
`chemical localization of thrombomodulin in the aqueous hu-
`mor passage of the rat eye. Histochem. Cell Biol. 108:121–131,
`1997.
`27. van der Zypen, E. Experimental morphological study on
`structure and function of the filtration angel of the rat eye.
`Ophthalmologica 174:285–298, 1997.
`28. Reme, C., Urner, U., and Aeberhard, B. The development of
`the chamber angle in the rat eye. Morphological character-
`istics of developmental stages. Graefe’s Arch. Clin. Exp. Oph-
`thalmol. 220:139–153, 1983.
`29. Nucci, P., Tredici, G., Manitto, M.P., et al. Neuron-specific
`enolase and embryology of the trabecular meshwork of the
`rat eye: An immunohistochemical study. Int. J. Biol. Markers
`7:253–255, 1992.
`30. Husain, S., Whitlock, N.A., Rice, D.S., et al. Effects of la-
`tanoprost on rodent intraocular pressure. Exp. Eye Res.
`83:1453–1458, 2006.
`31. Giuffre, G. The effects of prostaglandin F2alpha in the hu-
`man eye. Graefe’s Arch. Clin. Exp. Ophthalmol. 222:139–141,
`1985.
`32. Toris, C.B., Camras, C.B., and Yablonski, M.E. Effects of
`PhXA41, a new prostaglandin F2alpha analog, on aqueous
`humor dynamics in human eyes. Ophthalmology 100:
`1297–1304, 1993.
`33. Richter, M., and Krauss, A.H. Morphological changes in
`the anterior eye segment after long-term treatment with
`different receptor selective prostaglandin agonists and a
`prostamide. Invest. Ophthalmol. Vis. Sci. 44:4419–4426,
`2003.
`34. Sagara, T., Gaton, D.D., Lindsey, J.D., et al. Reduction of col-
`lagen type I in the ciliary muscle of inflamed monkey eyes.
`Invest. Ophthalmol. Vis. Sci. 40:2568–2576, 1999.
`35. Grobelny, D., Poncz, L., and Galardy, R.E. Inhibition of hu-
`man
`skin
`fibroblast
`collagenase,
`thermolysin, and
`Pseudomonas aeruginosa elastase by peptide hydroxamic
`acids. Biochemistry 31:7152–7154, 1992.
`36. Hao, J.L., Nagano, T., Nakamura, M., et al. Effect of galardin
`on collagen degradation by Pseudomonas aeruginosa. Exp. Eye
`Res. 69:595–601, 1999.
`37. Agren, M.S., Mirastschijski, U., Karlsmark, T., et al. Topical
`synthetic inhibitor of matrix metalloproteinases delays epi-
`dermal regeneration of human wounds. Exp. Dermatol.
`10:337–348, 2001.
`38. Bradley, J.M., Anderssohn, A.M., Colvis, C.M., et al. Medi-
`ation of laser trabeculoplasty-induced matrix metallopro-
`teinase expression by IL-1beta and TNFalpha. Invest. Oph-
`thalmol. Vis. Sci. 41:422–430, 2000.
`39. Parshley, D.E., Bradley, J.M., Fisk, A., et al. Laser trabecu-
`loplasty induces stromelysin expression by trabecular jux-
`tacanalicular cells. Invest. Ophthalmol. Vis. Sci. 37:795–804,
`1996.
`40. Raje, N., and Anderson, K. Thalidomide—a revival story. N.
`Engl. J. Med. 341:1565–1571, 1999.
`41. Tseng, S., Pak, G., Washenik, K., et al. Rediscovering thalido-
`mide: A review of its mechanism of action, side effects, and
`potential uses. J. Am. Acad. Dermatol. 35:969–979, 1996.
`42. Moreira, A.L., Sampaio, E.P., Zmuidzinas, A., et al. Thalido-
`mide exerts its inhibitory action on tumor necrosis factor al-
`
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`372
`
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`
`pha by enhancing mRNA degradation. J. Exp. Med.
`177:1675–1680, 1993.
`43. Corral, L.G., Haslett, P.A., Muller, G.W., et al. Differential
`cytokine modulation and T-cell activation by two distinct
`classes of thalidomide analogues that are potent inhibitors
`of TNF-a. J. Immunol. 163:380–386, 1999.
`44. Dinslage, S., Hueber, A., Diestelhorst, M., et al. The influ-
`ence of latanoprost 0.005% on aqueous humor flow and out-
`flow facility in glaucoma patients: A double-masked,
`placebo-controlled clinical study. Graefe’s Arch. Clin. Exp.
`Ophthalmol. 242:654–660, 2004.
`45. Toris, C.B., Zhan, G., Fan, S., et al. Effects of travoprost on
`aqueous humor dynamics in patients with elevated intraoc-
`ular pressure. J. Glaucoma. 16:189–195, 2007.
`
`46. Kamphuis, W., Schneemann, A., van Beek, L.M., et al.
`Prostanoid receptor gene expression profile in human tra-
`becular meshwork: A quantitative real-time PCR approach.
`Invest. Ophthalmol. Vis. Sci. 42:3209–3215, 2001.
`
`Received: March 28, 2008
`Accepted: March 31, 2008
`
`Reprint Requests: Shahid Husain
`Department of Ophthalmology
`Storm Eye Institute
`Medical University of South Carolina
`167 Ashley Avenue
`Charleston, SC 29425
`
`E-mail: husain@musc.edu
`
`
`
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