Topical application of L-arginine blocks advanced glycation by ascorbic acid in the lens of hSVCT2 transgenic mice

Xingjun Fan, Liu Xiaoqin, Breshey Potts, Christopher M. Strauch, Ina Nemet, Vincent M. Monnier

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Purpose: Previous experiments from our laboratory showed that the oral intake of selected guanidino compounds could block the formation of crystallin-bound advanced ascorbylation products. Here we tested whether these were also active when applied as eye drops. Methods: Two month old hSVCT2 transgenic mice (n=10) were treated twice daily with one drop of 0.1% L-arginine, γ-guanidinobutyric acid (GBA), penicillamine (PA) or N-acetylcysteine (NAC) in one eye and vehicle only in the other eye. After seven months, lens crystallins were isolated, dialyzed, and proteolytically digested to determine the proteinbound fluorescence at 335/385 and 370/440 nm excitation/emission and the advanced glycation/ascorbylation endproducts carboxymethyl-lysine (CML), carboxyethyl-lysine (CEL), glucosepane, glyoxal, and methylglyoxal hydroimidazolones G-H1 and MG-H1. The topical uptake of L-arginine and NAC was also evaluated in vitro and in vivo in rabbit lens. Results: In hSVCT2 mice, L-arginine decreased 335/385 and 370/440 nm fluorescence by 40% (p<0.001), CML, CEL, and glucosepane crystallin crosslinks by 35% (p<0.05), 30% (p<0.05), and 37% (p<0.05), respectively, without affecting MG-H1 and G-H1. NAC decreased 335/385 nm fluorescence by 50% (p<0.001) but, like PA and GBA, had no effect on other modifications. L-Arginine uptake into rabbit eyes treated topically reached identical lenticular plateau levels (~400 nmol/g wet weight) at 0.5% and 2.0% but levels remained three times higher at 5 h at 2% versus 0.5% concentration, respectively. In vitro studies showed a 100 fold higher L-arginine level than NAC levels, implicating high affinity uptake of the former. Conclusions: L-Arginine when applied both orally and topically is a potent and broad suppressor of advanced ascorbylation in the lens. Its uptake in rabbit lens upon topical application suggests transcorneal uptake into the human lens should be feasible for testing its potential anticataract properties in clinical trials.

Original languageEnglish (US)
Pages (from-to)2221-2227
Number of pages7
JournalMolecular Vision
Volume17
StatePublished - Sep 26 2011

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Transgenic Mice
Lenses
Ascorbic Acid
Arginine
Acetylcysteine
Crystallins
Penicillamine
Fluorescence
Rabbits
Lysine
Glyoxal
Pyruvaldehyde
Acids
Ophthalmic Solutions
Clinical Trials
Weights and Measures
N(6)-carboxymethyllysine
glucosepane
In Vitro Techniques

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Fan, X., Xiaoqin, L., Potts, B., Strauch, C. M., Nemet, I., & Monnier, V. M. (2011). Topical application of L-arginine blocks advanced glycation by ascorbic acid in the lens of hSVCT2 transgenic mice. Molecular Vision, 17, 2221-2227.

Topical application of L-arginine blocks advanced glycation by ascorbic acid in the lens of hSVCT2 transgenic mice. / Fan, Xingjun; Xiaoqin, Liu; Potts, Breshey; Strauch, Christopher M.; Nemet, Ina; Monnier, Vincent M.

In: Molecular Vision, Vol. 17, 26.09.2011, p. 2221-2227.

Research output: Contribution to journalArticle

Fan, X, Xiaoqin, L, Potts, B, Strauch, CM, Nemet, I & Monnier, VM 2011, 'Topical application of L-arginine blocks advanced glycation by ascorbic acid in the lens of hSVCT2 transgenic mice', Molecular Vision, vol. 17, pp. 2221-2227.
Fan, Xingjun ; Xiaoqin, Liu ; Potts, Breshey ; Strauch, Christopher M. ; Nemet, Ina ; Monnier, Vincent M. / Topical application of L-arginine blocks advanced glycation by ascorbic acid in the lens of hSVCT2 transgenic mice. In: Molecular Vision. 2011 ; Vol. 17. pp. 2221-2227.
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abstract = "Purpose: Previous experiments from our laboratory showed that the oral intake of selected guanidino compounds could block the formation of crystallin-bound advanced ascorbylation products. Here we tested whether these were also active when applied as eye drops. Methods: Two month old hSVCT2 transgenic mice (n=10) were treated twice daily with one drop of 0.1{\%} L-arginine, γ-guanidinobutyric acid (GBA), penicillamine (PA) or N-acetylcysteine (NAC) in one eye and vehicle only in the other eye. After seven months, lens crystallins were isolated, dialyzed, and proteolytically digested to determine the proteinbound fluorescence at 335/385 and 370/440 nm excitation/emission and the advanced glycation/ascorbylation endproducts carboxymethyl-lysine (CML), carboxyethyl-lysine (CEL), glucosepane, glyoxal, and methylglyoxal hydroimidazolones G-H1 and MG-H1. The topical uptake of L-arginine and NAC was also evaluated in vitro and in vivo in rabbit lens. Results: In hSVCT2 mice, L-arginine decreased 335/385 and 370/440 nm fluorescence by 40{\%} (p<0.001), CML, CEL, and glucosepane crystallin crosslinks by 35{\%} (p<0.05), 30{\%} (p<0.05), and 37{\%} (p<0.05), respectively, without affecting MG-H1 and G-H1. NAC decreased 335/385 nm fluorescence by 50{\%} (p<0.001) but, like PA and GBA, had no effect on other modifications. L-Arginine uptake into rabbit eyes treated topically reached identical lenticular plateau levels (~400 nmol/g wet weight) at 0.5{\%} and 2.0{\%} but levels remained three times higher at 5 h at 2{\%} versus 0.5{\%} concentration, respectively. In vitro studies showed a 100 fold higher L-arginine level than NAC levels, implicating high affinity uptake of the former. Conclusions: L-Arginine when applied both orally and topically is a potent and broad suppressor of advanced ascorbylation in the lens. Its uptake in rabbit lens upon topical application suggests transcorneal uptake into the human lens should be feasible for testing its potential anticataract properties in clinical trials.",
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AU - Xiaoqin, Liu

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AU - Strauch, Christopher M.

AU - Nemet, Ina

AU - Monnier, Vincent M.

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N2 - Purpose: Previous experiments from our laboratory showed that the oral intake of selected guanidino compounds could block the formation of crystallin-bound advanced ascorbylation products. Here we tested whether these were also active when applied as eye drops. Methods: Two month old hSVCT2 transgenic mice (n=10) were treated twice daily with one drop of 0.1% L-arginine, γ-guanidinobutyric acid (GBA), penicillamine (PA) or N-acetylcysteine (NAC) in one eye and vehicle only in the other eye. After seven months, lens crystallins were isolated, dialyzed, and proteolytically digested to determine the proteinbound fluorescence at 335/385 and 370/440 nm excitation/emission and the advanced glycation/ascorbylation endproducts carboxymethyl-lysine (CML), carboxyethyl-lysine (CEL), glucosepane, glyoxal, and methylglyoxal hydroimidazolones G-H1 and MG-H1. The topical uptake of L-arginine and NAC was also evaluated in vitro and in vivo in rabbit lens. Results: In hSVCT2 mice, L-arginine decreased 335/385 and 370/440 nm fluorescence by 40% (p<0.001), CML, CEL, and glucosepane crystallin crosslinks by 35% (p<0.05), 30% (p<0.05), and 37% (p<0.05), respectively, without affecting MG-H1 and G-H1. NAC decreased 335/385 nm fluorescence by 50% (p<0.001) but, like PA and GBA, had no effect on other modifications. L-Arginine uptake into rabbit eyes treated topically reached identical lenticular plateau levels (~400 nmol/g wet weight) at 0.5% and 2.0% but levels remained three times higher at 5 h at 2% versus 0.5% concentration, respectively. In vitro studies showed a 100 fold higher L-arginine level than NAC levels, implicating high affinity uptake of the former. Conclusions: L-Arginine when applied both orally and topically is a potent and broad suppressor of advanced ascorbylation in the lens. Its uptake in rabbit lens upon topical application suggests transcorneal uptake into the human lens should be feasible for testing its potential anticataract properties in clinical trials.

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