Pentosidine induces PDGF-B chain expression in ARPE-19 RPE cells

J. T. Handa, K. M. Reiser, Leonard M Hjelmeland

Research output: Contribution to journalArticle

Abstract

Purpose. Advanced glycation endproducts (AGEs) have been implicated in a number of aging complications such as altered matrix deposition. This alteration may be influenced by the abnormal expression of cytokines. PDGF, expressed by RPE cells, is known to influence matrix deposition and remodeling. The purpose of this study is to determine the effect of pentosidine, a known in vivo AGE, on the expression of PDGF-B chain in the human ARPE-19 RPE cell line. Methods. Pentosidine was synthesized from L-arginine, L-lysine, and Dribose, and purified. ARPE-19 cells were grown to confluence, rendered quiet by serum starvation, and exposed to pentosidine (0.01-lOfiM) for 1-24 hours. Total RNA was extracted, and mRNA expression of PDGF A and B chain was determined by Northern analysis. Steady state mRNA levels of PDGF A and B chain after pentosidine treatment were compared to unstimulated cells. PDGF-B mRNA levels were quantitated by autoradiography, and normalized against actin mRNA. Results, The maximum induction of steady state PDGF-B mRNA occurred after a 2 hour (1.75-fold increase) and 4 hour exposure (2.25-fold increase) to IjiM pentosidine whereas shorter (1 hour) and longer (8, 24 hour) pentosidine exposures did not induce PDGF-B mRNA. A 4 hour exposure to pentosidine (0.01-10|iM) induced PDGF-B mRNA from 1.5-6-fold over baseline in a dose dependent fashion. PDGF-A mRNA was not detected in ARPE-19 cells. Conclusions. Pentosidine, a well characterized AGE, altered the expression of PDGF-B chain in ARPE-19 cells. These findings suggest a role for AGEs in ocular diseases associated with aeine such as aee-related macular degeneration.

Original languageEnglish (US)
JournalInvestigative Ophthalmology and Visual Science
Volume38
Issue number4
StatePublished - 1997

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Messenger RNA
Eye Diseases
pentosidine
Macular Degeneration
Starvation
Autoradiography
Lysine
Arginine
Actins
RNA
Cytokines
Cell Line
Serum
platelet-derived growth factor A

ASJC Scopus subject areas

  • Ophthalmology

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Pentosidine induces PDGF-B chain expression in ARPE-19 RPE cells. / Handa, J. T.; Reiser, K. M.; Hjelmeland, Leonard M.

In: Investigative Ophthalmology and Visual Science, Vol. 38, No. 4, 1997.

Research output: Contribution to journalArticle

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abstract = "Purpose. Advanced glycation endproducts (AGEs) have been implicated in a number of aging complications such as altered matrix deposition. This alteration may be influenced by the abnormal expression of cytokines. PDGF, expressed by RPE cells, is known to influence matrix deposition and remodeling. The purpose of this study is to determine the effect of pentosidine, a known in vivo AGE, on the expression of PDGF-B chain in the human ARPE-19 RPE cell line. Methods. Pentosidine was synthesized from L-arginine, L-lysine, and Dribose, and purified. ARPE-19 cells were grown to confluence, rendered quiet by serum starvation, and exposed to pentosidine (0.01-lOfiM) for 1-24 hours. Total RNA was extracted, and mRNA expression of PDGF A and B chain was determined by Northern analysis. Steady state mRNA levels of PDGF A and B chain after pentosidine treatment were compared to unstimulated cells. PDGF-B mRNA levels were quantitated by autoradiography, and normalized against actin mRNA. Results, The maximum induction of steady state PDGF-B mRNA occurred after a 2 hour (1.75-fold increase) and 4 hour exposure (2.25-fold increase) to IjiM pentosidine whereas shorter (1 hour) and longer (8, 24 hour) pentosidine exposures did not induce PDGF-B mRNA. A 4 hour exposure to pentosidine (0.01-10|iM) induced PDGF-B mRNA from 1.5-6-fold over baseline in a dose dependent fashion. PDGF-A mRNA was not detected in ARPE-19 cells. Conclusions. Pentosidine, a well characterized AGE, altered the expression of PDGF-B chain in ARPE-19 cells. These findings suggest a role for AGEs in ocular diseases associated with aeine such as aee-related macular degeneration.",
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AU - Handa, J. T.

AU - Reiser, K. M.

AU - Hjelmeland, Leonard M

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Y1 - 1997

N2 - Purpose. Advanced glycation endproducts (AGEs) have been implicated in a number of aging complications such as altered matrix deposition. This alteration may be influenced by the abnormal expression of cytokines. PDGF, expressed by RPE cells, is known to influence matrix deposition and remodeling. The purpose of this study is to determine the effect of pentosidine, a known in vivo AGE, on the expression of PDGF-B chain in the human ARPE-19 RPE cell line. Methods. Pentosidine was synthesized from L-arginine, L-lysine, and Dribose, and purified. ARPE-19 cells were grown to confluence, rendered quiet by serum starvation, and exposed to pentosidine (0.01-lOfiM) for 1-24 hours. Total RNA was extracted, and mRNA expression of PDGF A and B chain was determined by Northern analysis. Steady state mRNA levels of PDGF A and B chain after pentosidine treatment were compared to unstimulated cells. PDGF-B mRNA levels were quantitated by autoradiography, and normalized against actin mRNA. Results, The maximum induction of steady state PDGF-B mRNA occurred after a 2 hour (1.75-fold increase) and 4 hour exposure (2.25-fold increase) to IjiM pentosidine whereas shorter (1 hour) and longer (8, 24 hour) pentosidine exposures did not induce PDGF-B mRNA. A 4 hour exposure to pentosidine (0.01-10|iM) induced PDGF-B mRNA from 1.5-6-fold over baseline in a dose dependent fashion. PDGF-A mRNA was not detected in ARPE-19 cells. Conclusions. Pentosidine, a well characterized AGE, altered the expression of PDGF-B chain in ARPE-19 cells. These findings suggest a role for AGEs in ocular diseases associated with aeine such as aee-related macular degeneration.

AB - Purpose. Advanced glycation endproducts (AGEs) have been implicated in a number of aging complications such as altered matrix deposition. This alteration may be influenced by the abnormal expression of cytokines. PDGF, expressed by RPE cells, is known to influence matrix deposition and remodeling. The purpose of this study is to determine the effect of pentosidine, a known in vivo AGE, on the expression of PDGF-B chain in the human ARPE-19 RPE cell line. Methods. Pentosidine was synthesized from L-arginine, L-lysine, and Dribose, and purified. ARPE-19 cells were grown to confluence, rendered quiet by serum starvation, and exposed to pentosidine (0.01-lOfiM) for 1-24 hours. Total RNA was extracted, and mRNA expression of PDGF A and B chain was determined by Northern analysis. Steady state mRNA levels of PDGF A and B chain after pentosidine treatment were compared to unstimulated cells. PDGF-B mRNA levels were quantitated by autoradiography, and normalized against actin mRNA. Results, The maximum induction of steady state PDGF-B mRNA occurred after a 2 hour (1.75-fold increase) and 4 hour exposure (2.25-fold increase) to IjiM pentosidine whereas shorter (1 hour) and longer (8, 24 hour) pentosidine exposures did not induce PDGF-B mRNA. A 4 hour exposure to pentosidine (0.01-10|iM) induced PDGF-B mRNA from 1.5-6-fold over baseline in a dose dependent fashion. PDGF-A mRNA was not detected in ARPE-19 cells. Conclusions. Pentosidine, a well characterized AGE, altered the expression of PDGF-B chain in ARPE-19 cells. These findings suggest a role for AGEs in ocular diseases associated with aeine such as aee-related macular degeneration.

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